WO2015150465A2

WO2015150465A2 - Plants having increased tolerance to herbicides

Info

Publication number: WO2015150465A2
Application number: PCT/EP2015/057198
Authority: WO
Inventors: Raphael Aponte; Stefan Tresch; Johannes Hutzler; Thomas Mietzner; Jill Marie Paulik; Chad BROMMER; Anja Simon; Florian Vogt
Original assignee: Basf Se
Priority date: 2014-04-03
Filing date: 2015-04-01
Publication date: 2015-10-08
Also published as: WO2015150465A3

Abstract

The present invention refers to aplant or plant part comprising a polynucleotide encoding a wildtype or mutant transketolase polypeptide, the expression of said polynucleotide confers to the plant or plant part tolerance to transketolase-inhibiting herbicides.

Description

PLANTS HAVING INCREASED TOLERANCE TO HERBICIDES

FIELD OF THE INVENTION This application claims priority to US provisional applications number US 61/974466, filed on April 3, 2014, the content of which is incorporated herein by reference in its entirety.

The present invention relates in general to methods for conferring on plants agricultural level tolerance to herbicides. Particularly, the invention refers to plants having an increased tolerance to herbicides, more specifically to herbicides which inhibit the enzyme transketolase (TK). More specifically, the present invention relates to methods and plants obtained by mutagenesis and cross-breeding and transformation that have an increased tolerance to herbicides, particularly TK-inhibiting herbicides. BACKGROUND OF THE INVENTION

Transketolase (EC 2.2.1.1 ) is integral to both the Calvin cycle and the oxidative pentose phosphate pathway of higher-plant chloroplasts. In the Calvin cycle, it catalyses the transfer of a two-carbon ketol group from either D-fructose- 6-phosphate or D-sedoheptulose-7- phosphate to D-glyceraldehyde- 3-phosphate to yield D-xylulose-5- phosphate and either D- erythrose-4-phosphate or D-ribose-5-phosphate, respectively [Flechner et al., Plant

Molecular Biology 32: 475-484, 1996]. In the pentose phosphate pathway these reversible reactions proceed in the opposite direction. Transketolase (TK) is thiamine pyrophosphate- dependent and is a dimer of 74 kDa subunits [de la Haba et al., J Biol Chem 214 409-426 (1955); Murphy and Walker, Planta 155, 316-320 (1982)]. Yeast transketolase is one of several thiamin diphosphate dependent enzymes whose three-dimensional structures have been determined [Schenk et al., The International Journal of Biochemistry & Cell Biology 30 (1998) 1297-1318]. Together with mutational analysis these structural data have led to detailed understanding of thiamin diphosphate catalyzed reactions. In the homodimer transketolase the two catalytic sites, where dihydroxyethyl groups are transferred from ketose donors to aldose acceptors, are formed at the interface between the two subunits, where the thiazole and pyrimidine rings of thiamin diphosphate are bound. Transketolase is ubiquitous and more than 30 full-length sequences are known. The encoded protein sequences contain two motifs of high homology; one common to all thiamin diphosphate- dependent enzymes and the other a unique transketolase motif [Schenk et al., The

International Journal of Biochemistry & Cell Biology 30 (1998) 1297-1318]. Higher plants in general might possess chloroplast and cytosolic TK isoenzymes, and, additionally, TK activity was observed to be localized to the chloroplast, at least in spinach leaves [Flechner et al., Plant Molecular Biology 32: 475-484, 1996 and references contained therein]. Since products of the transketolase-catalysed reaction serve as precursors for a number of synthetic compounds this enzyme has been exploited for industrial applications. The inventors of the present invention have now surprisingly found that over-expression of wildtype and mutant transketolase forms confers in plants tolerance/resistance to particular classes of TK-inhibiting herbicides as compared to the non-transformed and/or non- mutagenized plants or plant cells, respectively. More specifically, the inventors of the present invention have found that TK expression confers tolerance/resistance to cornexistin and/or hydrocornexistin.

Cornexistin and hydroxycornexistin are natural products derived from the fungus

Paecilomyces divaricatus. Isolation of cornexistin from the cultures of Paecilomyces species was published as early as 1989 by the Sankyo research group. The Sankyo Corporation discovered cornexistin during the screening of biological extracts for herbicidal use

(JP2256602). Later work from the DOW Elanco group described identification of

hydroxycornexistin also produced in Paecilomyces variotii SANK 21086 (US00542478). Both, cornexistin and hydroxycornexistin are highly potent herbicides that have the unique quality of being harmless to corn plants. Because of this quality, both molecules have attracted research interest. Cornexistin showed good activity as a herbicide as well as relative inactivity towards corn plants. Cornexistin and hydroxycornexistin has been synthesized by chemical synthesis only as diastereomeres (Org. Biomol. Chem., 2008, 6, 4012-4025). Nine-membered carbocyclic structures in general are rare in nature and their synthesis as well as the genes involved in the synthesis is still unknown and not described.

The problem of the present invention can be seen as to the provision of novel traits by identifying target polypeptides, the manipulation of which makes plants tolerant to herbicides.

Three main strategies are available for making plants tolerant to herbicides, i.e. (1 ) detoxifying the herbicide with an enzyme which transforms the herbicide, or its active metabolite, into non-toxic products, such as, for example, the enzymes for tolerance to bromoxynil or to basta (EP242236, EP337899); (2) mutating the target enzyme into a functional enzyme which is less sensitive to the herbicide, or to its active metabolite, such as, for example, the enzymes for tolerance to glyphosate (EP293356, Padgette S. R. et al., J.Biol. Chem., 266, 33, 1991 ); or (3) overexpressing the sensitive enzyme so as to produce quantities of the target enzyme in the plant which are sufficient in relation to the herbicide, in view of the kinetic constants of this enzyme, so as to have enough of the functional enzyme available despite the presence of its inhibitor.

The problem is solved by the subject-matter of the present invention. SUMMARY OF THE INVENTION

Accordingly, in one aspect, the present invention provides a plant or plant part comprising a polynucleotide encoding a wildtype or mutated TK polypeptide, the expression of said polynucleotide confers to the plant or plant part tolerance to TK-inhibiting herbicides.

In some aspects, the present invention provides a seed capable of germination into a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In one aspect, the present invention provides a plant cell of or capable of regenerating a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides, wherein the plant cell comprises the polynucleotide operably linked to a promoter.

In another aspect, the present invention provides a plant cell comprising a polynucleotide operably linked to a promoter operable in a cell, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In other aspects, the present invention provides a plant product prepared from a plant or plant part comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In some aspects, the present invention provides a progeny or descendant plant derived from a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, wherein the progeny or descendant plant comprises in at least some of its cells the recombinant polynucleotide operably linked to the promoter, the expression of the wildtype or mutated TK polypeptide conferring to the progeny or descendant plant tolerance to the TK-inhibiting herbicides.

In other aspects, the present invention provides a method for controlling weeds at a locus for growth of a plant, the method comprising: (a) applying an herbicide composition comprising TK-inhibiting herbicides to the locus; and (b) planting a seed at the locus, wherein the seed is capable of producing a plant that comprises in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In some aspects, the present invention provides a method for controlling weeds at a locus for growth of a plant, the method comprising: applying an herbicidal composition comprising TK-inhibiting herbicides to the locus; wherein said locus is: (a) a locus that contains: a plant or a seed capable of producing said plant; or (b) a locus that is to be after said applying is made to contain the plant or the seed; wherein the plant or the seed comprises in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In one aspect, step (a) occurs before, after, or concurrently with step (b). In other aspects, the present invention provides a method of producing a plant having tolerance to TK-inhibiting herbicides, the method comprising regenerating a plant from a plant cell transformed with a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In one aspect, the present invention provides a method of producing a progeny plant having tolerance to TK-inhibiting herbicides, the method comprising: crossing a first TK-inhibiting herbicides- tolerant plant with a second plant to produce a TK-inhibiting herbicides- tolerant progeny plant, wherein the first plant and the progeny plant comprise in at least some of their cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In addition, the present invention refers to a method for identifying a TK-inhibiting herbicide by using a wild-type or mutated TK of the present invention encoded by a nucleic acid which comprises the nucleotide sequence of SEQ ID NO: 182 or 183, or a variant, homologue, paralogue or orthologue thereof.

Said method comprises the steps of:

a) generating a transgenic cell or plant comprising a nucleic acid encoding a mutated TK of the present invention, wherein the mutated TK of the present invention is expressed;

b) applying a TK-inhibiting herbicide to the transgenic cell or plant of a) and to a control cell or plant of the same variety;

c) determining the growth or the viability of the transgenic cell or plant and the control cell or plant after application of said test compound, and d) selecting test compounds which confer reduced growth to the control cell or plant as compared to the growth of the transgenic cell or plant.

Another object refers to a method of identifying a nucleotide sequence encoding a mutated TK which is resistant or tolerant to a TK-inhibiting herbicide, the method comprising:

a) generating a library of mutated TK-encoding nucleic acids,

b) screening a population of the resulting mutated TK-encoding nucleic acids by expressing each of said nucleic acids in a cell or plant and treating said cell or plant with a TK- inhibiting herbicide,

c) comparing the TK-inhibiting herbicide-tolerance levels provided by said population of mutated TK encoding nucleic acids with the TK-inhibiting herbicide-tolerance level provided by a control TK-encoding nucleic acid,

d) selecting at least one mutated TK-encoding nucleic acid that provides a significantly increased level of tolerance to a TK-inhibiting herbicide as compared to that provided by the control TK-encoding nucleic acid.

In a preferred embodiment, the mutated TK-encoding nucleic acid selected in step d) provides at least 2-fold as much tolerance to a TK-inhibiting herbicide as compared to that provided by the control TK-encoding nucleic acid.

The resistance or tolerance can be determined by generating a transgenic plant comprising a nucleic acid sequence of the library of step a) and comparing said transgenic plant with a control plant.

Another object refers to a method of identifying a plant or algae containing a nucleic acid encoding a mutated TK which is resistant or tolerant to a TK-inhibiting herbicide, the method comprising:

a) identifying an effective amount of a TK-inhibiting herbicide in a culture of plant cells or green algae.

b) treating said plant cells or green algae with a mutagenizing agent,

c) contacting said mutagenized cells population with an effective amount of TK-inhibiting herbicide, identified in a),

d) selecting at least one cell surviving these test conditions,

e) PCR-amplification and sequencing of TK genes from cells selected in d) and comparing such sequences to wild-type TK gene sequences, respectively.

In a preferred embodiment, the mutagenizing agent is ethylmethanesulfonate.

Another object refers to an isolated nucleic acid encoding a mutated TK, the nucleic acid comprising the sequence of SEQ ID NO: 182, or 183, or a variant thereof, as defined hereinafter.

In a preferred embodiment, the nucleic acid being identifiable by a method as defined above. Another object refers to an isolated mutated TK polypeptide, the polypeptide comprising the sequence set forth in SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 1 10, 1 1 1 , 1 12, 1 13, 1 14, 1 15, 1 16, 1 17, 1 18, 1 19, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , a variant, derivative, orthologue, paralogue or homologue thereof, as defined hereinafter.

In still further aspects, the present invention provides a plant or plant part comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides, wherein the plant or plant part further exhibits a second or third herbicide-tolerant trait. In another embodiment, the invention refers to a plant cell transformed by and expressing a wild-type or a mutated TK nucleic acid according to the present invention or a plant which has been mutated to obtain a plant expressing, preferably over-expressing a wild-type or a mutated TK nucleic acid according to the present invention, wherein expression of said nucleic acid in the plant cell results in increased resistance or tolerance to a TK -inhibiting herbicide as compared to a wild type variety of the plant cell

In another embodiment, the invention refers to a plant comprising a plant cell according to the present invention, wherein expression of the nucleic acid in the plant results in the plant's increased resistance to TK-inhibiting herbicide as compared to a wild type variety of the plant.

The plants of the present invention can be transgenic or non-transgenic.

Preferably, the expression of the nucleic acid of the invention in the plant results in the plant's increased resistance to TK-inhibiting herbicides as compared to a wild type variety of the plantin another embodiment, the invention refers to a seed produced by a transgenic plant comprising a plant cell of the present invention, wherein the seed is true breeding for an increased resistance to a TK-inhibiting herbicide as compared to a wild type variety of the seed.

In another embodiment, the invention refers to a method of producing a transgenic plant cell with an increased resistance to a TK-inhibiting herbicide as compared to a wild type variety of the plant cell comprising, transforming the plant cell with an expression cassette comprising a a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide.

In another embodiment, the invention refers to a method of producing a transgenic plant comprising, (a) transforming a plant cell with an expression cassette comprising a a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, and (b) generating a plant with an increased resistance to TK-inhibiting herbicide from the plant cell.

Preferably, the expression cassette further comprises a transcription initiation regulatory region and a translation initiation regulatory region that are functional in the plant.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1

A, B, C, D, E, F, G, and H show a Multiple alignment using clustalw of sequences 1 to 181 depicted below in Table 1 .

KEY TO SEQUENCE LISTING

Table 1

SEQ ID Organism / Origin / Accession number

1 1 [transketolase Spinacia oleracea]

2 transketolase [Spinacia oleracea]

3 gi|2529342|gb|AAD10219.11 transketolase [Spinacia oleracea]

4 gi|223548870|gb|EEF50359.1 | transketolase, putative [Ricinus communis]

5 Transketolase_1 _AMATU

6 Transketolase_2_AMATU

7 Transketolase_3_AMATU

8 Transketolase_1 _KOSCS

9 Transketolase_2_KOSCS

10 Transketolase_LEMPA

1 1 Partial_Sequence_Transketolase_ARBTH

gi|514760041 |ref|XP_004964445.1 | PREDICTED: transketolase, chloroplastic-

12 like [Setaria italica]

gi|568875658|ref|XP_006490909.1 | PREDICTED: transketolase-2,

13 chloroplastic-like [Citrus sinensis]

14 gi|351735634|gb|AEQ59483.1 | chloroplast transketolase [Cucumis sativus]

15 gi|1 18487947|gb|ABK95795.1 | [Populus trichocarpa]

gi|470142472|ref|XP_004306931 .1 | PREDICTED: [Fragaria vesca subsp.

16 vesca]

gi|561015858|gb|ESW14662.11 hypothetical protein PHAVU_007G006600g

17 [Phaseolus vulgaris]

18 gi|449498919|ref|XP_004160671.11 PREDICTED: transketolase, chloroplastic- like [Cucumis sativus]

gi|356576867|ref|XP_003556551.11 transketolase, chloroplastic [Glycine max] gi|469474163|gb|AGH33875.11 transketolase [Camellia sinensis]

gi|462416702|gb|EMJ21439.1 | [Prunus persica]

gi|222843004|gb|EEE80551 .11 transketolase family protein [Populus trichocarpa]

gi|460388792|ref|XP_004240048.11 transketolase, chloroplastic-like [Solanum lycopersicum]

gi|356536526|ref|XP_003536788.11 transketolase, chloroplastic-like [Glycine max]

gi|355516131 |gb|AES97754.11 Transketolase [Medicago truncatula] gi|330255441 |gb|AEC10535.11 transketolase [Arabidopsis thaliana] gi|1 18481093|gb|ABK92500.11 [Populus trichocarpa]

gi|449474170|ref|XP_004154093.1 | transketolase, chloroplastic-like [Cucumis sativus]

gi|565357366|ref|XP_006345515.11 transketolase, chloroplastic-like [Solanum tuberosum]

gi|502121526|ref|XP_004497357.11 transketolase, chloroplastic-like [Cicer arietinum]

Nicotianatabacum

Arabidopsisthaliana

Theobromacacao

Capsellarubella

Arabidopsisthaliana

Vitisvinifera

Solanumlycopersicum

Zeamays

Arabidopsisthaliana

gi|3559814|emb|CAA75777.11 transketolase [Capsicum annuum]

gi|568214657|ref|NP_001275202.11 transketolase [Solanum tuberosum] gi|356506190|ref|XP_003521870.11 transketolase, chloroplastic [Glycine max] gi|7329685|emb|CAB82679.1 | transketolase-like protein [Arabidopsis thaliana] gi|565465348|ref|XP_006290638.1 | [Capsella rubella]

gi|28190676|gb|AAO33154.11 transketolase [Oryza sativa]

gi|561026815|gb|ESW25455.1 | [Phaseolus vulgaris]

gi|1 15466224|ref|NP_00105671 1.11 Os06g0133800 [Oryza sativa]

gi|312282187|dbj|BAJ33959.1 | [Thellungiella halophila]

gi|573948603|ref|XP_006656605.1 | partial Transketolase [Oryza brachyantha] gi|548832450|gb|ERM95246.1 [Amborella trichopoda]

gi|548847307|gb|ERN06491.1 | [Amborella trichopoda]

gi|502146626|ref|XP_004506536.11 [Cicer arietinum]

gi|3571 10873|ref|XP_003557240.1 | [Brachypodium distachyon] gi|355506195|gb|AES87337.11 Transketolase [Medicago truncatula] gi|147835837|emb|CAN72939.1 | hypothetical protein [Vitis vinifera] gi|225454009|ref|XP_002280760.11 transketolase [Vitis vinifera]

gi|297318807|gb|EFH49229.11 transketolase [Arabidopsis lyrata subsp. lyrata] gi|241915985|gb|EER89129.11 [Sorghum bicolor]

gi|413953334|gb|AFW85983.11 transketolase isoform 1 [Zea mays] gi|224127366|ref|XP_002320056.1 | [Populus trichocarpa]

Hordeum vulgare subsp. vulgare

Picea sitchensis

Arabidopsis thaliana

Sorghum bicolor

Zea mays

Eutrema salsugineum

Polygonum tinctorium

Oryza sativa Japonica Group

Oryza sativa Indica Group

Oryza brachyantha

Setaria italica

Physcomitrella patens

Setaria italica

Physcomitrella patens

Oryza sativa Indica Group

gi|1 15457470|ref|NP_001052335.11 Os04g0266900 [Oryza sativa Japonica] gi|475481099|gb|EMT02862.11 Transketolase [Aegilops tauschii]

gi|357167367|ref|XP_003581 128.11 transketolase [Brachypodium distachyon] gi|125547501 |gb|EAY93323.1 | [Oryza sativa Indica Group]

gi|514760053|ref|XP_004964448.11 transketolase [Setaria italica]

gi|242072546|ref|XP_002446209.11 [Sorghum bicolor]

gi|514760061 |ref|XP_004964450.11 transketolase [Setaria italica]

gi|326515912|dbj|BAJ87979.1 | [Hordeum vulgare subsp. vulgare]

gi|474352176|gb|EMS63024.11 Transketolase [Triticum urartu]

gi|664903|emb|CAA86609.11 transketolase [Craterostigma plantagineum] gi|147788852|emb|CAN60522.1 | [Vitis vinifera]

gi|300140959|gb|EFJ07676.1 | [Selaginella moellendorffii]

gi|413953333|gb|AFW85982.1 | [Zea mays]

gi|414587814|tpg|DAA38385.11 [Zea mays]

gi|162683807|gb|EDQ70214.1 | [Physcomitrella patens]

gi|125595949|gb|EAZ35729.1 | [Oryza sativa Japonica Group]

gi|222423758|dbj|BAH19845.11 AT3G60750 [Arabidopsis thaliana]

Medicago truncatula

Sorghum bicolor

Volvox carteri f. nagariensis 96 Chlamydomonas reinhardtii

97 Geitlerinema sp. PCC 7407

98 Craterostigma plantagineum

99 Pleurocapsa sp. PCC 7327

100 Moorea producens 3L

101 Fischerella muscicola

102 Leptolyngbya sp. Heron Island J

103 Cyanothece sp. PCC 7425

104 Paecilomyces divaricatus (Transketolase, scaffold001 g03940)

105 SPTREMBL|A5DSN8_LODEL RecName: Full=Transketolase; EC=2.2.1 .1 ;

106 SPTREMBL|A8PWG5_MALGO RecName: Full=Transketolase; EC=2.2.1 .1 ;

107 SPTREMBL|B6K5F9_SCHJY RecName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|C0LQF6_PICPA RecName: Full=Transketolase; EC=2.2.1.1 ;

108 Flags: Fragment;

SPTREMBL|C7ZAM9_NECH7 SubName: Full=Putative uncharacterized

109 protein;

SPTREMBL|D7UPI2_OGAME SubName: Full=Dihydroxyacetone synthase;

1 10 EC=2.2.1 .3;

1 1 1 SPTREMBL|D7UPI3_OGAME SubName: Full=DAS-like protein;

SPTREMBL|D8Q788_SCHCM SubName: Full=Putative uncharacterized

1 12 protein;

1 13 SPTREMBL|F4P3U6_BATDJ RecName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|F5HCT5_CRYNB SubName: Full=Putative uncharacterized

1 14 protein;

SPTREMBL|G3BBY9_CANTC SubName: Full=Putative uncharacterized

1 15 protein;

1 16 SPTREMBL|G7DV49_MIXOS SubName: Full=Uncharacterized protein;

1 17 SPTREMBL|G8C0M6_TETPH RecName: Full=Transketolase; EC=2.2.1 .1 ;

1 18 SPTREMBL|G9MFC9_HYPVG SubName: Full=Uncharacterized protein;

1 19 SPTREMBL|H0GCH3_9SACH RecName: Full=Transketolase; EC=2.2.1 .1 ;

120 SPTREMBL|I 1 BXP2_RHI09 RecName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|I 1 RJN7_GIBZE SubName: Full=Dihydroxyacetone synthase;

121 SubName: Full=Uncharacterized protein;

122 SPTREMBL|I2K2H5_DEKBR RecName: Full=Transketolase; EC=2.2.1 .1 ;

123 SPTREMBL|J9N9S0_FUSO4 SubName: Full=Uncharacterized protein;

124 SPTREMBL|J9VL23_CRYNH RecName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|L0PF66_PNEJ8 SubName: Full=l WGS project CAKMOOOOOOOO

125 data, strain SE8, contig 242;

126 SPTREMBL|L2FB38_COLGN SubName: Full=Dihydroxyacetone synthase;

127 SPTREMBL|Q5KHG5_CRYNJ SubName: Full=Transketolase, putative;

128 SPTREMBL|Q6BN17_DEBHA SubName: Full=DEHA2F00968p;

129 SPTREMBL|Q6BXS5_DEBHA SubName: Full=DEHA2B00616p; 130 Paecilomyces divaricatus (Transketolase, scaffold020g00760)

131 SPTREMBL|A1 DJZ3_NEOFI SubName: Full=Transketolase;

132 SPTREMBL|A5DD66_PICGU SubName: Full=Putative uncharacterized protein;

133 SPTREMBL|A5DSN8_LODEL RecName: Full=Transketolase; EC=2.2.1 .1 ;

134 SPTREMBL|A8PWG5_MALGO RecName: Full=Transketolase; EC=2.2.1 .1 ;

135 SPTREMBL|B6K5F9_SCHJY RecName: Full=Transketolase; EC=2.2.1 .1 ;

136 SPTREMBL|B8M240_TALSN RecName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|B8M3Z5_TALSN SubName: Full=Transketolase, putative;

137 EC=2.2.1 .3;

SPTREMBL|C0LQF6_PICPA RecName: Full=Transketolase; EC=2.2.1.1 ;

138 Flags: Fragment;

SPTREMBL|C7ZAM9_NECH7 SubName: Full=Putative uncharacterized

139 protein;

SPTREMBL|D7UPI2_OGAME SubName: Full=Dihydroxyacetone synthase;

140 EC=2.2.1 .3;

141 SPTREMBL|D7UPI3_OGAME SubName: Full=DAS-like protein;

SPTREMBL|D8Q788_SCHCM SubName: Full=Putative uncharacterized

142 protein;

143 SPTREMBL|E6ZQ75_SPORE SubName: Full=Probable Transketolase;

SPTREMBL|F2QV92_PICP7 SubName: Full=Dihydroxyacetone synthase

144 variant 1 ; EC=2.2.1 .3;

145 SPTREMBL|F4P3U6_BATDJ RecName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|F5HCT5_CRYNB SubName: Full=Putative uncharacterized

146 protein;

147 SPTREMBL|F9GER6_FUSOF SubName: Full=Uncharacterized protein;

148 SPTREMBL|F9XCD0_MYCGM SubName: Full=Uncharacterized protein;

149 SPTREMBL|G2YJE4_BOTF4 SubName: Full=Similar to transketolase;

150 SPTREMBL|G3B8B6_CANTC SubName: Full=Dihydroxyacetone synthase;

SPTREMBL|G3BBY9_CANTC SubName: Full=Putative uncharacterized

151 protein;

152 SPTREMBL|G7DV49_MIXOS SubName: Full=Uncharacterized protein;

153 SPTREMBL|G8BNE8_TETPH RecName: Full=Transketolase; EC=2.2.1 .1 ;

154 SPTREMBL|G8C0M6_TETPH RecName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|H0EPU8_GLAL7 SubName: Full=Putative Dihydroxyacetone

155 synthase;

156 Paecilomyces divaricatus (Transketolase, scaffold025g01090)

157 SPTREMBL|A8NV31_COPC7 SubName: Full=Transketolase;

SPTREMBL|D8Q4R0_SCHCM SubName: Full=Putative uncharacterized

158 protein;

SPTREMBL|E3JQR8_PUCGT SubName: Full=Putative uncharacterized

159 protein;

160 SPTREMBL|F4RJP4_MELLP SubName: Full=Putative uncharacterized protein; 161 SPTREMBL|G7DVE0_MIXOS SubName: Full=Uncharacterized protein;

162 SPTREMBL|H0EEH2_GLAL7 SubName: Full=Putative Transketolase;

SPTREMBL|I4YAU9_WALSC SubName: Full=Thiamin diphosphate-binding

163 protein;

SPTREMBL|I4YE20_WALSC SubName: Full=Thiamin diphosphate-binding

164 protein;

165 SPTREMBL|K1WGM0_MARBU SubName: Full=Transketolase;

166 SPTREMBL|K5WES2_PHACS SubName: Full=Uncharacterized protein;

SPTREMBL|Q55T38_CRYNB SubName: Full=Putative uncharacterized

167 protein;

168 SPTREMBL|L1J2M8_GUITH SubName: Full=Uncharacterized protein;

169 SPTREMBL|A0YNC4_LYNSP SubName: Full=Uncharacterized protein;

170 SPTREMBL|A0ZLZ6_NODSP SubName: Full=Transketolase; EC=2.2.1 .1 ;

SPTREMBL|B4WLI3_9SYNE SubName: Full=Transketolase, C-terminal

171 domain protein;

SPTREMBL|C9M6X8_9BACT SubName: Full=Transketolase, C-terminal

172 domain protein;

173 SPTREMBL|D1 Y3C7_9BACT SubName: Full=Transketolase; EC=2.2.1 .1 ;

174 SPTREMBL|D2Z2G4_9BACT SubName: Full=Transketolase central region;

SPTREMBL|D3L2W3_9BACT SubName: Full=Transketolase, thiamine

175 diphosphate binding domain protein;

176 SPTREMBL|D6THL7_9CHLR SubName: Full=Transketolase domain protein;

SPTREMBL|D8F7T0_9DELT SubName: Full=Putative 1 -deoxy-D-xylulose-5-

177 phosphate synthase;

178 SPTREMBL|F4XPA7_9CYAN SubName: Full=Transketolase;

SPTREMBL|G2H8B8_9DELT SubName: Full=Transketolase, C-terminal

179 domain protein;

SPTREMBL|G3J1 Y8_9GAMM SubName: Full=Transketolase domain-

180 containing protein;

SPTREMBL|G9PZY8_9BACT SubName: Full=Putative uncharacterized

181 protein;

DETAILED DESCRIPTION

The articles "a" and "an" are used herein to refer to one or more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one or more elements.

As used herein, the word "comprising," or variations such as "comprises" or "comprising," will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. The term "control of undesired vegetation or weeds" is to be understood as meaning the killing of weeds and/or otherwise retarding or inhibiting the normal growth of the weeds. Weeds, in the broadest sense, are understood as meaning all those plants which grow in locations where they are undesired. The weeds of the present invention include, for example, dicotyledonous and monocotyledonous weeds. Dicotyledonous weeds include, but are not limited to, weeds of the genera: Sinapis, Lepidium, Galium, Stellaria, Matricaria, Anthemis, Galinsoga, Chenopodium, Urtica, Senecio, Amaranthus, Portulaca, Xanthium, Convolvulus, Ipomoea, Polygonum, Sesbania, Ambrosia, Cirsium, Carduus, Sonchus, Solanum, Rorippa, Rotala, Lindernia, Lamium, Veronica, Abutilon, Emex, Datura, Viola, Galeopsis, Papaver, Centaurea, Trifolium, Ranunculus, and Taraxacum. Monocotyledonous weeds include, but are not limited to, weeds of of the genera: Echinochloa, Setaria,

Panicum, Digitaria, Phleum, Poa, Festuca, Eleusine, Brachiaria, Lolium, Bromus, Avena, Cyperus, Sorghum, Agropyron, Cynodon, Monochoria, Fimbristyslis, Sagittaria, Eleocharis, Scirpus, Paspalum, Ischaemum, Sphenoclea, Dactyloctenium, Agrostis, Alopecurus, and Apera. In addition, the weeds of the present invention can include, for example, crop plants that are growing in an undesired location. For example, a volunteer maize plant that is in a field that predominantly comprises soybean plants can be considered a weed, if the maize plant is undesired in the field of soybean plants. The term "plant" is used in its broadest sense as it pertains to organic material and is intended to encompass eukaryotic organisms that are members of the Kingdom Plantae, examples of which include but are not limited to vascular plants, vegetables, grains, flowers, trees, herbs, bushes, grasses, vines, ferns, mosses, fungi and algae, etc, as well as clones, offsets, and parts of plants used for asexual propagation (e.g. cuttings, pipings, shoots, rhizomes, underground stems, clumps, crowns, bulbs, corms, tubers, rhizomes,

plants/tissues produced in tissue culture, etc.). The term "plant" further encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, shoots, stems, leaves, roots (including tubers), flowers, florets, fruits, pedicles, peduncles, stamen, anther, stigma, style, ovary, petal, sepal, carpel, root tip, root cap, root hair, leaf hair, seed hair, pollen grain, microspore, cotyledon, hypocotyl, epicotyl, xylem, phloem, parenchyma, endosperm, a companion cell, a guard cell, and any other known organs, tissues, and cells of a plant, and tissues and organs, wherein each of the aforementioned comprise the gene/nucleic acid of interest. The term "plant" also encompasses plant cells, suspension cultures, callus tissue, embryos, meristematic regions, gametophytes, sporophytes, pollen and microspores, again wherein each of the aforementioned comprises the gene/nucleic acid of interest.

Plants that are particularly useful in the methods of the invention include all plants which belong to the superfamily Viridiplantae, in particular monocotyledonous and dicotyledonous plants including fodder or forage legumes, ornamental plants, food crops, trees or shrubs selected from the list comprising Acer spp., Actinidia spp., Abelmoschus spp., Agave sisalana, Agropyron spp., Agrostis stolonifera, Allium spp., Amaranthus spp., Ammophila arenaria, Ananas comosus, Annona spp., Apium graveolens, Arachis spp, Artocarpus spp., Asparagus officinalis, Avena spp. (e.g. Avena sativa, Avena fatua, Avena byzantina, Avena fatua var. sativa, Avena hybrida), Averrhoa carambola, Bambusa sp., Benincasa hispida, Bertholletia excelsea, Beta vulgaris, Brassica spp. (e.g. Brassica napus, Brassica rapa ssp. [canola, oilseed rape, turnip rape]), Cadaba farinosa, Camellia sinensis, Canna indica, Cannabis sativa, Capsicum spp., Carex elata, Carica papaya, Carissa macrocarpa, Carya spp., Carthamus tinctorius, Castanea spp., Ceiba pentandra, Cichorium endivia,

Cinnamomum spp., Citrullus lanatus, Citrus spp., Cocos spp., Coffea spp., Colocasia esculenta, Cola spp., Corchorus sp., Coriandrum sativum, Corylus spp., Crataegus spp., Crocus sativus, Cucurbita spp., Cucumis spp., Cynara spp., Daucus carota, Desmodium spp., Dimocarpus longan, Dioscorea spp., Diospyros spp., Echinochloa spp., Elaeis (e.g. Elaeis guineensis, Elaeis oleifera), Eleusine coracana, Eragrostis tef, Erianthus sp., Eriobotrya japonica, Eucalyptus sp., Eugenia uniflora, Fagopyrum spp., Fagus spp., Festuca arundinacea, Ficus carica, Fortunella spp., Fragaria spp., Ginkgo biloba, Glycine spp. (e.g. Glycine max, Soja hispida or Soja max), Gossypium hirsutum, Helianthus spp. (e.g. Helianthus annuus), Hemerocallis fulva, Hibiscus spp., Hordeum spp. (e.g. Hordeum vulgare), Ipomoea batatas, Juglans spp., Lactuca sativa, Lathyrus spp., Lens culinaris, Linum usitatissimum, Litchi chinensis, Lotus spp., Luffa acutangula, Lupinus spp., Luzula sylvatica, Lycopersicon spp. (e.g. Lycopersicon esculentum, Lycopersicon lycopersicum, Lycopersicon pyriforme), Macrotyloma spp., Malus spp., Malpighia emarginata, Mammea americana, Mangifera indica, Manihot spp., Manilkara zapota, Medicago sativa, Melilotus spp., Mentha spp., Miscanthus sinensis, Momordica spp., Morus nigra, Musa spp.,

Nicotiana spp., Olea spp., Opuntia spp., Ornithopus spp., Oryza spp. (e.g. Oryza sativa, Oryza latifolia), Panicum miliaceum, Panicum virgatum, Passiflora edulis, Pastinaca sativa, Pennisetum sp., Persea spp., Petroselinum crispum, Phalaris arundinacea, Phaseolus spp., Phleum pratense, Phoenix spp., Phragmites australis, Physalis spp., Pinus spp., Pistacia vera, Pisum spp., Poa spp., Populus spp., Prosopis spp., Prunus spp., Psidium spp., Punica granatum, Pyrus communis, Quercus spp., Raphanus sativus, Rheum rhabarbarum, Ribes spp., Ricinus communis, Rubus spp., Saccharum spp., Salix sp., Sambucus spp., Secale cereale, Sesamum spp., Sinapis sp., Solanum spp. (e.g. Solanum tuberosum, Solanum integrifolium or Solanum lycopersicum), Sorghum bicolor, Spinacia spp.,

Syzygium spp., Tagetes spp., Tamarindus indica, Theobroma cacao, Trifolium spp., Tripsacum dactyloides, Triticosecale rimpaui, Triticum spp. (e.g. Triticum aestivum, Triticum durum, Triticum turgidum, Triticum hybernum, Triticum macha, Triticum sativum, Triticum monococcum or Triticum vulgare), Tropaeolum minus, Tropaeolum majus, Vaccinium spp., Vicia spp., Vigna spp., Viola odorata, Vitis spp., Zea mays, Zizania palustris, Ziziphus spp., amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, strawberry, sugar beet, sugar cane, sunflower, tomato, squash, tea and algae, amongst others. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include inter alia soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato or tobacco. Further preferebly, the plant is a monocotyledonous plant, such as sugarcane. Further preferably, the plant is a cereal, such as rice, maize, wheat, barley, millet, rye, sorghum or oats.

Generally, the term "herbicide" is used herein to mean an active ingredient that kills, controls or otherwise adversely modifies the growth of plants. The preferred amount or concentration of the herbicide is an "effective amount" or "effective concentration." By "effective amount" and "effective concentration" is intended an amount and concentration, respectively, that is sufficient to kill or inhibit the growth of a similar, wild-type, plant, plant tissue, plant cell, or host cell, but that said amount does not kill or inhibit as severely the growth of the herbicide-resistant plants, plant tissues, plant cells, and host cells of the present invention. Typically, the effective amount of a herbicide is an amount that is routinely used in agricultural production systems to kill weeds of interest. Such an amount is known to those of ordinary skill in the art. Herbicidal activity is exhibited by herbicides useful for the the present invention when they are applied directly to the plant or to the locus of the plant at any stage of growth or before planting or emergence. The effect observed depends upon the plant species to be controlled, the stage of growth of the plant, the application parameters of dilution and spray drop size, the particle size of solid components, the environmental conditions at the time of use, the specific compound employed, the specific adjuvants and carriers employed, the soil type, and the like, as well as the amount of chemical applied. These and other factors can be adjusted as is known in the art to promote non-selective or selective herbicidal action. Generally, it is preferred to apply the herbicide postemergence to relatively immature undesirable vegetation to achieve the maximum control of weeds. By a "herbicide-tolerant" or "herbicide-resistant" plant, it is intended that a plant that is tolerant or resistant to at least one herbicide at a level that would normally kill, or inhibit the growth of, a normal or wild-type plant. By "herbicide-tolerant wildtype or mutated TK protein" or "herbicide -resistant wildtype or mutated TK protein", it is intended that such a TK protein displays higher TK activity, relative to the TK activity of a wild-type TK protein, when in the presence of at least one herbicide that is known to interfere with TK activity and at a

concentration or level of the herbicide that is known to inhibit the TK activity of the wild-type mutated TK protein. Furthermore, the TK activity of such a herbicide-tolerant or herbicide- resistant mutated TK protein may be referred to herein as "herbicide-tolerant" or "herbicide- resistant" TK activity.

Levels of herbicide that normally inhibit growth of a non-tolerant plant are known and readily determined by those skilled in the art. Examples include the amounts recommended by manufacturers for application. The maximum rate is an example of an amount of herbicide that would normally inhibit growth of a non-tolerant plant. For the present invention, the terms "herbicide-tolerant" and "herbicide-resistant" are used interchangeably and are intended to have an equivalent meaning and an equivalent scope. Similarly, the terms "herbicide-tolerance" and "herbicide-resistance" are used interchangeably and are intended to have an equivalent meaning and an equivalent scope. Similarly, the terms "tolerant" and "resistant" are used interchangeably and are intended to have an equivalent meaning and an equivalent scope. As used herein, in regard to an herbicidal composition useful in various embodiments hereof, terms such as TK-inhibiting herbicides, and the like, refer to those agronomically acceptable herbicide active ingredients (A.I.) recognized in the art. Similarly, terms such as fungicide, nematicide, pesticide, and the like, refer to other agronomically acceptable active ingredients recognized in the art.

When used in reference to a particular mutant enzyme or polypeptide, terms such as herbicide-tolerant and herbicide-tolerance refer to the ability of such enzyme or polypeptide to perform its physiological activity in the presence of an amount of an herbicide A.I. that would normally inactivate or inhibit the activity of the wild-type (non-mutant) version of said enzyme or polypeptide. For example, when used specifically in regard to a TK enzyme, it refers specifically to the ability to tolerate a TK-inhibitor. By "herbicide-tolerant wildtype or mutated TK protein" or "herbicide -resistant wildtype or mutated TK protein", it is intended that such a TK protein displays higher TK activity, relative to the TK activity of a wild-type TK protein, when in the presence of at least one herbicide that is known to interfere with TK activity and at a concentration or level of the herbicide that is known to inhibit the TK activity of the wild-type or mutated TK protein. Furthermore, the TK activity of such a herbicide- tolerant or herbicide-resistant wildtype or mutated TK protein may be referred to herein as "herbicide-tolerant" or "herbicide-resistant" TK activity.

As used herein, "recombinant," when referring to nucleic acid or polypeptide, indicates that such material has been altered as a result of human application of a recombinant technique, such as by polynucleotide restriction and ligation, by polynucleotide overlap- extension, or by genomic insertion or transformation. A gene sequence open reading frame is recombinant if that nucleotide sequence has been removed from it natural text and cloned into any type of artificial nucleic acid vector. The term recombinant also can refer to an organism having a recombinant material, e.g., a plant that comprises a recombinant nucleic acid can be considered a recombinant plant.

The term "transgenic plant" refers to a plant that comprises a heterologous polynucleotide. Preferably, the heterologous polynucleotide is stably integrated within the genome such that the polynucleotide is passed on to successive generations. The heterologous

polynucleotide may be integrated into the genome alone or as part of a recombinant expression cassette. "Transgenic" is used herein to refer to any cell, cell line, callus, tissue, plant part or plant, the genotype of which has been so altered by the presence of heterologous nucleic acid including those transgenic organisms or cells initially so altered, as well as those created by crosses or asexual propagation from the initial transgenic organism or cell. In some embodiments, a "recombinant" organism is a "transgenic" organism. The term "transgenic" as used herein is not intended to encompass the alteration of the genome (chromosomal or extra-chromosomal) by conventional plant breeding methods (e.g., crosses) or by naturally occurring events such as, e.g., self-fertilization, random cross-fertilization, non-recombinant viral infection, non-recombinant bacterial transformation, non- recombinant transposition, or spontaneous mutation. As used herein, "mutagenized" refers to an organism or DNA thereof having alteration(s) in the biomolecular sequence of its native genetic material as compared to the sequence of the genetic material of a corresponding wild-type organism or DNA, wherein the

alteration(s) in genetic material were induced and/or selected by human action. Examples of human action that can be used to produce a mutagenized organism or DNA include, but are not limited to, as illustrated in regard to herbicide tolerance: tissue culture of plant cells (e.g., calli) and selection thereof with herbicides (e.g., TK-inhibiting herbicides), treatment of plant cells with a chemical mutagen such as EMS and subsequent selection with

herbicide(s); or by treatment of plant cells with x-rays and subsequent selection with herbicide(s). Any method known in the art can be used to induce mutations. Methods of inducing mutations can induce mutations in random positions in the genetic material or can induce mutations in specific locations in the genetic material (i.e., can be directed mutagenesis techniques), such as by use of a genoplasty technique.

As used herein, a "genetically modified organism" (GMO) is an organism whose genetic characteristics contain alteration(s) that were produced by human effort causing

transfection that results in transformation of a target organism with genetic material from another or "source" organism, or with synthetic or modified-native genetic material, or an organism that is a descendant thereof that retains the inserted genetic material. The source organism can be of a different type of organism (e.g., a GMO plant can contain bacterial genetic material) or from the same type of organism (e.g., a GMO plant can contain genetic material from another plant). As used herein in regard to plants and other organisms, "recombinant," "transgenic," and "GMO" are considered synonyms and indicate the presence of genetic material from a different source; in contrast, "mutagenized" is used to refer to a plant or other organism, or the DNA thereof, in which no such transgenic material is present, but in which the native genetic material has become mutated so as to differ from a corresponding wild-type organism or DNA.

As used herein, "wild-type" or "corresponding wild-type plant" means the typical form of an organism or its genetic material, as it normally occurs, as distinguished from, e.g., mutagenized and/or recombinant forms. Similarly, by "control cell" or "similar, wild-type, plant, plant tissue, plant cell or host cell" is intended a plant, plant tissue, plant cell, or host cell, respectively, that lacks the herbicide-resistance characteristics and/or particular polynucleotide of the invention that are disclosed herein. The use of the term "wild-type" is not, therefore, intended to imply that a plant, plant tissue, plant cell, or other host cell lacks recombinant DNA in its genome, and/or does not possess herbicide-resistant

characteristics that are different from those disclosed herein. As used herein, "descendant" refers to any generation plant. In some embodiments, a descendant is a first, second, third, fourth, fifth, sixth, seventh, eight, ninth, or tenth generation plant. As used herein, "progeny" refers to a first generation plant.

The term "seed" comprises seeds of all types, such as, for example, true seeds, caryopses, achenes, fruits, tubers, seedlings and similar forms. In the context of Brassica and Sinapis species, "seed" refers to true seed(s) unless otherwise specified. For example, the seed can be seed of transgenic plants or plants obtained by traditional breeding methods.

Examples of traditional breeding methods can include cross-breeding, selfing, back- crossing, embryo rescue, in-crossing, out-crossing, inbreeding, selection, asexual propagation, and other traditional techniques as are known in the art. Although exemplified with reference to specific plants or plant varieties and their hybrids, in various embodiments, the presently described methods using TK-inhibiting herbicides can be employed with a variety of commercially valuable plants. TK-inhibiting herbicides-tolerant plant lines described as useful herein can be employed in weed control methods either directly or indirectly, i. e. either as crops for herbicide treatment or as TK-inhibiting herbicides-tolerance trait donor lines for development, as by traditional plant breeding, to produce other varietal and/or hybrid crops containing such trait or traits. All such resulting variety or hybrids crops, containing the ancestral TK-inhibiting herbicides-tolerance trait or traits can be referred to herein as progeny or descendant of the ancestral, TK-inhibiting herbicides-tolerant line(s). Such resulting plants can be said to retain the "herbicide tolerance characteristic(s)" of the ancestral plant, i.e. meaning that they possess and express the ancestral genetic molecular components responsible for the trait.

In one aspect, the present invention provides a plant or plant part comprising a

polynucleotide encoding a wildtype or mutated TK polypeptide, the expression of said polynucleotide confers to the plant or plant part tolerance to TK-inhibiting herbicides.

In a preferred embodiment, the plant has been previously produced by a process comprising recombinantly preparing a plant by introducing and over-expressing a wild-type or mutated TK transgene according to the present invention, as described in greater detail hereinfter.

In another preferred embodiment, the plant has been previously produced by a process comprising in situ mutagenizing plant cells, to obtain plant cells which express a mutated TK.

In another embodiment, the polynucleotide encoding the wildtype or mutated TK

polypeptide polypeptide comprises the nucleic acid sequence set forth in SEQ ID NO: 182 or 183 a variant or derivative thereof. In other embodiments, the wildtype or mutated TK polypeptide for use according to the present invention is a functional variant having, over the full-length of the variant, at least about 80%, illustratively, at least about 80%, 90%, 95%, 98%, 99% or more amino acid sequence identity to SEQ ID NO: 1,2, 3,4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181.

In another embodiment, the wildtype or mutated TK polypeptide for use according to the present invention is a functional fragment of a polypeptide having the amino acid sequence set forth in SEQ ID NO: 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181.

It is recognized that the TK polynucleotide molecules and TK polypeptides of the invention encompass polynucleotide molecules and polypeptides comprising a nucleotide or an amino acid sequence that is sufficiently identical to nucleotide sequences set forth in SEQ ID Nos: 182 or 183, or to the amino acid sequences set forth in SEQ ID Nos: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181. The term "sufficiently identical" is used herein to refer to a first amino acid or nucleotide sequence that contains a sufficient or minimum number of identical or equivalent (e.g., with a similar side chain) amino acid residues or nucleotides to a second amino acid or nucleotide sequence such that the first and second amino acid or nucleotide sequences have a common structural domain and/or common functional activity. Generally, "sequence identity" refers to the extent to which two optimally aligned DNA or amino acid sequences are invariant throughout a window of alignment of components, e.g., nucleotides or amino acids. An "identity fraction" for aligned segments of a test sequence and a reference sequence is the number of identical components that are shared by the two aligned sequences divided by the total number of components in reference sequence segment, i.e., the entire reference sequence or a smaller defined part of the reference sequence. "Percent identity" is the identity fraction times 100. Optimal alignment of sequences for aligning a comparison window are well known to those skilled in the art and may be conducted by tools such as the local homology algorithm of Smith and Waterman, the homology alignment algorithm of Needleman and Wunsch, the search for similarity method of Pearson and Lipman, and preferably by computerized implementations of these algorithms such as GAP, BESTFIT, FASTA, and TFASTA available as part of the GCG. Wisconsin Package. (Accelrys Inc. Burlington, Mass.) Polynucleotides and Oligonucleotides

By an "isolated polynucleotide", including DNA, RNA, or a combination of these, single or double stranded, in the sense or antisense orientation or a combination of both, dsRNA or otherwise, we mean a polynucleotide which is at least partially separated from the polynucleotide sequences with which it is associated or linked in its native state. Preferably, the isolated polynucleotide is at least 60% free, preferably at least 75% free, and most preferably at least 90% free from other components with which they are naturally

associated. As the skilled addressee would be aware, an isolated polynucleotide can be an exogenous polynucleotide present in, for example, a transgenic organism which does not naturally comprise the polynucleotide. Furthermore, the terms "polynucleotide(s)", "nucleic acid sequence(s)", "nucleotide sequence(s)", "nucleic acid(s)", "nucleic acid molecule" are used interchangeably herein and refer to nucleotides, either ribonucleotides or

deoxyribonucleotides or a combination of both, in a polymeric unbranched form of any length.

The term "mutated TK nucleic acid" refers to a TK nucleic acid having a sequence that is mutated from a wild-type TK nucleic acid and that confers increased TK-inhibiting herbicide tolerance to a plant in which it is expressed. Furthermore, the term "mutated transketolase (mutated TK)" refers to the replacement of an amino acid of the wild-type primary sequences of SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 1 1 1 , 112, 1 13, 114, 1 15, 116, 1 17, 1 18, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , or a variant, a derivative, a homologue, an orthologue, or paralogue thereof, with another amino acid. The expression "mutated amino acid" will be used below to designate the amino acid which is replaced by another amino acid, thereby designating the site of the mutation in the primary sequence of the protein.

In a preferred embodiment, the TK nucleotide sequence encoding a mutated TK comprises the sequence of SEQ ID NO: 182, or 183, or a variant or derivative thereof Furthermore, it will be understood by the person skilled in the art that the TK nucleotide sequences encompasse homologues, paralogues and and orthologues of SEQ ID NO: 182 or 183, as defined hereinafter.

The term "variant" with respect to a sequence (e.g., a polypeptide or nucleic acid sequence such as - for example - a transcription regulating nucleotide sequence of the invention) is intended to mean substantially similar sequences. For nucleotide sequences comprising an open reading frame, variants include those sequences that, because of the degeneracy of the genetic code, encode the identical amino acid sequence of the native protein. Naturally occurring allelic variants such as these can be identified with the use of well-known molecular biology techniques, as, for example, with polymerase chain reaction (PCR) and hybridization techniques. Variant nucleotide sequences also include synthetically derived nucleotide sequences, such as those generated, for example, by using site-directed mutagenesis and for open reading frames, encode the native protein comprising the sequence of SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 1 1 1 , 1 12, 1 13, 114, 1 15, 116, 1 17, 1 18, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , as well as those that encode a polypeptide having amino acid substitutions relative to the native protein, e.g. the mutated TK according to the present invention as disclosed herein.

Generally, nucleotide sequence variants of the invention will have at least 30, 40, 50, 60, to 70%, e.g., preferably 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%, to 79%, generally at least 80%, e.g., 81 %-84%, at least 85%, e.g., 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, to 98% and 99% nucleotide "sequence identity" to the nucleotide sequence of SEQ ID NO: 182 or 183. The % identity of a polynucleotide is determined by GAP (Needleman and Wunsch, 1970) analysis (GCG program) with a gap creation penalty=5, and a gap extension penalty=0.3. Unless stated otherwise, the query sequence is at least 45 nucleotides in length, and the GAP analysis aligns the two sequences over a region of at least 45 nucleotides. Preferably, the query sequence is at least 150 nucleotides in length, and the GAP analysis aligns the two sequences over a region of at least 150 nucleotides. More preferably, the query sequence is at least 300 nucleotides in length and the GAP analysis aligns the two sequences over a region of at least 300 nucleotides. Even more preferably, the GAP analysis aligns the two sequences over their entire length.

Polypeptides

By "substantially purified polypeptide" or "purified" a polypeptide is meant that has been separated from one or more lipids, nucleic acids, other polypeptides, or other contaminating molecules with which it is associated in its native state. It is preferred that the substantially purified polypeptide is at least 60% free, more preferably at least 75% free, and more preferably at least 90% free from other components with which it is naturally associated. As the skilled addressee will appreciate, the purified polypeptide can be a recombinantly produced polypeptide. The terms "polypeptide" and "protein" are generally used

interchangeably and refer to a single polypeptide chain which may or may not be modified by addition of non-amino acid groups. It would be understood that such polypeptide chains may associate with other polypeptides or proteins or other molecules such as co-factors. The terms "proteins" and "polypeptides" as used herein also include variants, mutants, modifications, analogous and/or derivatives of the polypeptides of the invention as described herein.

The % identity of a polypeptide is determined by GAP (Needleman and Wunsch, 1970) analysis (GCG program) with a gap creation penalty=5, and a gap extension penalty=0.3. The query sequence is at least 25 amino acids in length, and the GAP analysis aligns the two sequences over a region of at least 25 amino acids. More preferably, the query sequence is at least 50 amino acids in length, and the GAP analysis aligns the two sequences over a region of at least 50 amino acids. More preferably, the query sequence is at least 100 amino acids in length and the GAP analysis aligns the two sequences over a region of at least 100 amino acids. Even more preferably, the query sequence is at least 250 amino acids in length and the GAP analysis aligns the two sequences over a region of at least 250 amino acids. Even more preferably, the GAP analysis aligns the two sequences over their entire length. With regard to a defined polypeptide, it will be appreciated that % identity figures higher than those provided above will encompass preferred embodiments. Thus, where applicable, in light of the minimum % identity figures, it is preferred that the TK polypeptide of the invention comprises an amino acid sequence which is at least 40%, more preferably at least 45%, more preferably at least 50%, more preferably at least 55%, more preferably at least 60%, more preferably at least 65%, more preferably at least 70%, more preferably at least 75%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 91 %, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, more preferably at least 99%, more preferably at least 99.1 %, more preferably at least 99.2%, more preferably at least 99.3%, more preferably at least 99.4%, more preferably at least 99.5%, more preferably at least 99.6%, more preferably at least 99.7%, more preferably at least 99.8%, and even more preferably at least 99.9% identical to SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 1 10, 11 1 , 1 12, 113, 1 14, 1 15, 116, 1 17, 118, 1 19, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181.

By "variant" polypeptide is intended a polypeptide derived from the protein of SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 1 1 1 , 112, 1 13, 1 14, 115, 1 16, 117, 1 18, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , by deletion (so-called truncation) or addition of one or more amino acids to the N-terminal and/or C-terminal end of the native protein; deletion or addition of one or more amino acids at one or more sites in the native protein; or substitution of one or more amino acids at one or more sites in the native protein Such variants may result from, for example, genetic polymorphism or from human manipulation. Methods for such manipulations are generally known in the art.

"Derivatives" of a protein encompass peptides, oligopeptides, polypeptides, proteins and enzymes having amino acid substitutions, deletions and/or insertions relative to the unmodified protein in question and having similar biological and functional activity as the unmodified protein from which they are derived. Thus, functional variants and fragments of the TK polypeptides, and nucleic acid molecules encoding them, also are within the scope of the present invention, and unless specifically described otherwise, irrespective of the origin of said polypeptide and irrespective of whether it occurs naturally. Various assays for functionality of a TK polypeptide can be employed. For example, a functional variant or fragment of the TK polypeptide can be assayed to determine its ability to confer TK- inhibiting herbicides detoxification. By way of illustration, a TK-inhibiting herbicides detoxification rate can be defined as a catalytic rate sufficient to provide a determinable increase in tolerance to TK-inhibiting herbicides in a plant or plant part comprising a recombinant polynucleotide encoding the variant or fragment of the TK polypeptide, wherein the plant or plant part expresses the variant or fragment at up to about 0.5%, illustratively, about 0.05 to about 0.5%, about 0.1 to about 0.4%, and about 0.2 to about 0.3%, of the total cellular protein relative to a similarly treated control plant that does not express the variant or fragment.

In a preferred embodiment, the wildtype or mutated TK polypeptide is a functional variant or fragment of a transketolase having the amino acid sequence set forth in SEQ ID NO: 1 or 2, wherein the functional variant or fragment has at least about 80% amino acid sequence identity to SEQ ID NO:1 or 2. For the avoidance of doubts, SEQ ID NO: 2 is identical to SEQ ID NO: 1 except that SEQ ID NO: 2 lacks the N-terminal transit peptide comprising amino acids 1-73 of SEQ ID NO: 1 [MAASSSLSTL SHHQTLLSHP KTHLPTTPAS SLLVPTTSSK VNGVLLKSTS SSRRLRVGSA SAVVRAAAVE ALE] (see Table 2a and 2b herein below for corresponding amino acid residues)

In other embodiments, the functional variant or fragment further has a TK-inhibiting herbicides detoxification rate defined as a catalytic rate sufficient to provide a determinable increase in tolerance to TK-inhibiting herbicides in a plant or plant part comprising a recombinant polynucleotide encoding the variant or fragment, wherein the plant or plant part expresses the variant or fragment at up to about 0.5% of the total cellular protein to a similarly treated control plant that does not express the variant or fragment. "Homologues" of a protein encompass peptides, oligopeptides, polypeptides, proteins and enzymes having amino acid substitutions, deletions and/or insertions relative to the unmodified protein in question and having similar biological and functional activity as the unmodified protein from which they are derived. In addition, one of ordinary skill in the art will further appreciate that changes can be introduced by mutation into the nucleotide sequences of the invention thereby leading to changes in the amino acid sequence of the encoded proteins without altering the biological activity of the proteins. Thus, for example, an isolated polynucleotide molecule encoding a mutated TK polypeptide having an amino acid sequence that differs from that of SEQ ID NO: 1 or 2 can be created by introducing one or more nucleotide substitutions, additions, or deletions into the corresponding nucleotide sequence, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced by standard techniques, such as site-directed mutagenesis and PCR- mediated mutagenesis. Such variant nucleotide sequences are also encompassed by the present invention. For example, preferably, conservative amino acid substitutions may be made at one or more predicted preferably nonessential amino acid residues. A

"nonessential" amino acid residue is a residue that can be altered from the wild-type sequence of a protein without altering the biological activity, whereas an "essential" amino acid residue is required for biological activity.

A deletion refers to removal of one or more amino acids from a protein.

An insertion refers to one or more amino acid residues being introduced into a

predetermined site in a protein. Insertions may comprise N-terminal and/or C-terminal fusions as well as intra-sequence insertions of single or multiple amino acids. Generally, insertions within the amino acid sequence will be smaller than N- or C-terminal fusions, of the order of about 1 to 10 residues. Examples of N- or C-terminal fusion proteins or peptides include the binding domain or activation domain of a transcriptional activator as used in the yeast two-hybrid system, phage coat proteins, (histidine)-6-tag, glutathione S- transferase-tag, protein A, maltose-binding protein, dihydrofolate reductase, Tag^»100 epitope, c-myc epitope, FLAG^®-epitope, lacZ, CMP (calmodulin-binding peptide), HA epitope, protein C epitope and VSV epitope.

A substitution refers to replacement of amino acids of the protein with other amino acids having similar properties (such as similar hydrophobicity, hydrophilicity, antigenicity, propensity to form or break a-helical structures or β-sheet structures). Amino acid

substitutions are typically of single residues, but may be clustered depending upon functional constraints placed upon the polypeptide and may range from 1 to 10 amino acids; insertions will usually be of the order of about 1 to 10 amino acid residues. A conservative amino acid substitution is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Such substitutions would not be made for conserved amino acid residues, or for amino acid residues residing within a conserved motif. Conservative substitution tables are well known in the art (see for example Creighton (1984) Proteins. W.H. Freeman and Company (Eds). Amino acid substitutions, deletions and/or insertions may readily be made using peptide synthetic techniques well known in the art, such as solid phase peptide synthesis and the like, or by recombinant DNA manipulation. Methods for the manipulation of DNA sequences to produce substitution, insertion or deletion variants of a protein are well known in the art. For example, techniques for making substitution mutations at predetermined sites in DNA are well known to those skilled in the art and include M13 mutagenesis, T7-Gen in vitro mutagenesis (USB, Cleveland, OH), QuickChange Site Directed mutagenesis (Stratagene, San Diego, CA), PCR-mediated site-directed mutagenesis or other site-directed mutagenesis protocols.

"Derivatives" further include peptides, oligopeptides, polypeptides which may, compared to the amino acid sequence of the naturally-occurring form of the protein, such as the protein of interest, comprise substitutions of amino acids with non-naturally occurring amino acid residues, or additions of non-naturally occurring amino acid residues. "Derivatives" of a protein also encompass peptides, oligopeptides, polypeptides which comprise naturally occurring altered (glycosylated, acylated, prenylated, phosphorylated, myristoylated, sulphated etc.) or non-naturally altered amino acid residues compared to the amino acid sequence of a naturally-occurring form of the polypeptide. A derivative may also comprise one or more non-amino acid substituents or additions compared to the amino acid sequence from which it is derived, for example a reporter molecule or other ligand, covalently or non-covalently bound to the amino acid sequence, such as a reporter molecule which is bound to facilitate its detection, and non-naturally occurring amino acid residues relative to the amino acid sequence of a naturally-occurring protein. Furthermore, "derivatives" also include fusions of the naturally-occurring form of the protein with tagging peptides such as FLAG, HIS6 or thioredoxin (for a review of tagging peptides, see Terpe, Appl. Microbiol. Biotechnol. 60, 523-533, 2003). "Orthologues" and "paralogues" encompass evolutionary concepts used to describe the ancestral relationships of genes. Paralogues are genes within the same species that have originated through duplication of an ancestral gene; orthologues are genes from different organisms that have originated through speciation, and are also derived from a common ancestral gene. A non-limiting list of examples of such orthologues is shown in Table 1. It will be understood by the person skilled in the art that the sequences of SEQ ID NOs:2-181 as listed in Table 1 represent orthologues and paralogues to SEQ ID NO:1.

It is well-known in the art that paralogues and orthologues may share distinct domains harboring suitable amino acid residues at given sites, such as binding pockets for particular substrates or binding motifs for interaction with other proteins.

The term "domain" refers to a set of amino acids conserved at specific positions along an alignment of sequences of evolutionarily related proteins. While amino acids at other positions can vary between homologues, amino acids that are highly conserved at specific positions indicate amino acids that are likely essential in the structure, stability or function of a protein. Identified by their high degree of conservation in aligned sequences of a family of protein homologues, they can be used as identifiers to determine if any polypeptide in question belongs to a previously identified polypeptide family. The term "motif or "consensus sequence" refers to a short conserved region in the sequence of evolutionarily related proteins. Motifs are frequently highly conserved parts of domains, but may also include only part of the domain, or be located outside of conserved domain (if all of the amino acids of the motif fall outside of a defined domain).

Specialist databases exist for the identification of domains, for example, SMART (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864; Letunic et al. (2002) Nucleic Acids Res 30, 242-244), InterPro (Mulder et al., (2003) Nucl. Acids. Res. 31 , 315-318), Prosite (Bucher and Bairoch (1994), A generalized profile syntax for biomolecular sequences motifs and its function in automatic sequence interpretation. (In) ISMB-94; Proceedings 2nd International Conference on Intelligent Systems for Molecular Biology. Altman R., Brutlag D., Karp P., Lathrop R., Searls D., Eds., pp53-61 , AAAI Press, Menlo Park; Hulo et al., Nucl. Acids. Res. 32:D134-D137, (2004)), or Pfam (Bateman et al., Nucleic Acids Research 30(1 ): 276-280 (2002)). A set of tools for in silico analysis of protein sequences is available on the ExPASy proteomics server (Swiss Institute of Bioinformatics (Gasteiger et al., ExPASy: the proteomics server for in-depth protein knowledge and analysis, Nucleic Acids Res. 31 :3784-3788(2003)). Domains or motifs may also be identified using routine techniques, such as by sequence alignment.

Methods for the alignment of sequences for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol 48: 443-453) to find the global (i.e. spanning the complete sequences) alignment of two sequences that maximizes the number of matches and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The software for performing BLAST analysis is publicly available through the National Centre for Biotechnology Information (NCBI).

Homologues may readily be identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83), with the default pairwise alignment parameters, and a scoring method in percentage (See Figure 1 ). Global percentages of similarity and identity may also be determined using one of the methods available in the MatGAT software package (Campanella et al., BMC Bioinformatics. 2003 Jul 10;4:29. MatGAT: an application that generates similarity/identity matrices using protein or DNA sequences.). Minor manual editing may be performed to optimise alignment between conserved motifs, as would be apparent to a person skilled in the art. Furthermore, instead of using full-length sequences for the identification of homologues, specific domains may also be used. The sequence identity values may be determined over the entire nucleic acid or amino acid sequence or over selected domains or conserved motif(s), using the programs mentioned above using the default parameters. For local alignments, the Smith-Waterman algorithm is particularly useful (Smith TF, Waterman MS (1981 ) J. Mol. Biol 147(1 );195-7).

The proteins of the invention may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions. Methods for such manipulations are generally known in the art. For example, amino acid sequence variants can be prepared by mutations in the DNA. Methods for mutagenesis and nucleotide sequence alterations are well known in the art. See, for example, Kunkel (1985) PNAS, 82:488-492; Kunkel et al. (1987) Methods in Enzymol. 154:367-382; U.S. Patent No. 4,873,192; Walker and Gaastra, eds. (1983) Techniques in Molecular Biology (MacMillan Publishing Company, New York) and the references cited therein. Guidance as to appropriate amino acid substitutions that do not affect biological activity of the protein of interest may be found in the model of

Dayhoff et al. (1978) Atlas of Protein Sequence and Structure (Natl. Biomed. Res. Found., Washington, D. C), herein incorporated by reference. Conservative substitutions, such as exchanging one amino acid with another having similar properties, may be preferable. Alternatively, variant nucleotide sequences can be made by introducing mutations randomly along all or part of a coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened to identify mutants that encode proteins that retain activity. For example, following mutagenesis, the encoded protein can be expressed recombinantly, and the activity of the protein can be determined using standard assay techniques.

The inventors of the present invention have found that by substituting one or more of the key amino acid residues of the TK enzyme of SEQ ID NO: 1 or 2, e.g. by employing one of the above described methods to mutate the TK encoding nucleic acids, the tolerance or resistance to particular TK-inhibiting herbicides could be remarkably increased Preferred substitutions of mutated TK are those that increase the herbicide tolerance of the plant, but leave the biological activitiy of the oxidase activity substantially unaffected.

Accordingly, in another object of the present invention refers to a TK polypeptide, comprising the sequence of SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 1 1 1 , 1 12, 1 13, 114, 1 15, 116, 1 17, 1 18, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , a variant, derivative, orthologue, paralogue or homologue thereof, the key amino acid residues of which is substituted by any other amino acid.

It will be understood by the person skilled in the art that amino acids located in a close proximity to the positions of amino acids mentioned below may also be substituted. Thus, in another embodiment the variant of SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 1 10, 1 1 1 , 1 12, 1 13, 1 14, 1 15, 1 16, 1 17, 1 18, 1 19, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , a variant, derivative, orthologue, paralogue or homologue thereof comprises a mutated TK, wherein an amino acid ±3, ±2 or ±1 amino acid positions from a key amino acid is substituted by any other amino acid.

Based on techniques well-known in the art, a highly characteristic sequence pattern can be developed, by means of which further of mutated TK candidates with the desired activity may be searched.

Searching for further mutated TK candidates by applying a suitable sequence pattern would also be encompassed by the present invention. It will be understood by a skilled reader that the present sequence pattern is not limited by the exact distances between two adjacent amino acid residues of said pattern. Each of the distances between two neighbours in the above patterns may, for example, vary independently of each other by up to ±10, ± 5, ±3, ±2 or ±1 amino acid positions without substantially affecting the desired activity. Furthermore, by applying the method of site directed mutagenesis, in particular saturation mutagenes (see e.g. Schenk et al., Biospektrum 03/2006, pages 277-279), the inventors of the present invention have identified and generated specific amino acid subsitutions and

combinations thereof, which - when introduced into a plant by transforming and expressing the respective mutated TK encoding nucleic acid - confer increased herbicide resistance or tolerance to a TK inhibiting herbicide to said plant.

Thus, in a particularly preferred embodiment, the variant or derivative of the mutated TK refers to a TK polypeptide comprising SEQ ID NO: 1 , a orthologue, paralogue, or homologue thereof, wherein the amino acid sequence differs from the wildtype amino acid sequence of a TK polypeptide at one or more positions corresponding to the following positions of SEQ ID NO:1 : 265, 267, 337, 342, 343, 458, 459, 460, 461 , 463, 51 1 , 512, 513, 514, 515, 544.

Examples of differences at these amino acid positions include, but are not limited to, one or more of the following:

the amino acid at or corresponding to position 265 is other than isoleucine;

the amino acid at or corresponding to position 267 is other than isoleucine;

the amino acid at or corresponding to position 337 is other than tyrosine;

the amino acid at or corresponding to position 342 is other than serine;

the amino acid at or corresponding to position 343 is other than alanine;

the amino acid at or corresponding to position 458 is other than leucine;

the amino acid at or corresponding to position 459 is other than alanine; the amino acid at or corresponding to position 460 s other than serine;

the amino acid at or corresponding to position 461 s other than serine;

the amino acid at or corresponding to position 463 s other than methionine;

the amino acid at or corresponding to position 51 1 s other than Threonine;

the amino acid at or corresponding to position 512 s other than phenylalanine;

the amino acid at or corresponding to position 513 s other than phenylalanine;

the amino acid at or corresponding to position 514 s other than valine;

the amino acid at or corresponding to position 515 s other than phenylalanine

the amino acid at or corresponding to position 544 s other than leucine;

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 265 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, Met, Phe, Tyr, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 267 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, Met, Phe, Tyr, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 337 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Phe, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Arg, His, Lys, Asp, Glu, Thr, Asn, Gin Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Phe, Tyr, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 343 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Val, Leu, lie, Met, Phe, Tyr, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 458 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, lie, Met, Phe, Tyr, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 459 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Val, Leu, lie, Met, Phe, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 460 is Arg, His, Lys, Asp, Glu, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Phe, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 461 is Arg, His, Lys, Asp, Glu, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Phe, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Phe, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 51 1 is Arg, His, Lys, Asp, Glu, Ser, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Phe, Met, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 512 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 513 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 514 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Leu, lie, Phe, Met, Tyr, or Trp. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 515 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Tyr, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 544 is Arg, His, Lys, Asp, Glu, Ser, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, lie, Met, Phe, Tyr, or Trp.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Arg, His, Lys, Asp, Glu, Thr, Asn, Gin, Cys, Gly, Pro, Ala, Val, Leu, lie, Met, Phe, Tyr, or Trp, and the amino acid at or

corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Arg, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is His, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Lys, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Asp, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Glu, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Thr, and the amino acid at or corresponding to position 343 is Pro. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Asn, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Gin, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Cys, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Gly, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Pro, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Ala, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Val, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Leu, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is lie, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Met, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Phe, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Tyr, and the amino acid at or corresponding to position 343 is Pro.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Trp, and the amino acid at or corresponding to position 343 is Pro.

corresponding to position 343 is Gly. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Arg, and the amino acid at or corresponding to position 343 is Gly. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is His, and the amino acid at or corresponding to position 343 is Gly. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Lys, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Asp, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Glu, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Thr, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Asn, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Gin, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Cys, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Gly, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Pro, and the amino acid at or corresponding to position 343 is Gly. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Ala, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Val, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Leu, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is lie, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Met, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Phe, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Tyr, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 342 is Trp, and the amino acid at or corresponding to position 343 is Gly.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Val, Cys, Ala, Ser, Gly, and the amino acid at or corresponding to position 544 is Thr, Ala, Ser, Cys, Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Val, and the amino acid at or corresponding to position 544 is Thr.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Val, and the amino acid at or corresponding to position 544 is Ala.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Val, and the amino acid at or corresponding to position 544 is Ser.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Val, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Val, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 544 is Thr.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 544 is Ala.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 544 is Ser. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ala, and the amino acid at or corresponding to position 544 is Thr.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ala, and the amino acid at or corresponding to position 544 is Ala.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ala, and the amino acid at or corresponding to position 544 is Ser.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ala, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ala, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ser, and the amino acid at or corresponding to position 544 is Thr.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ser, and the amino acid at or corresponding to position 544 is Ala.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ser, and the amino acid at or corresponding to position 544 is Ser.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ser, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Ser, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 544 is Thr.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 544 is Ala.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 544 is Ser.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 544 is Val. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, Cys, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Thr, Cys, Ser, Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Thr.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Ser. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Thr. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Ser.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, Cys, and the amino acid at or corresponding to position 511 is Ser, and the amino acid at or corresponding to position 544 is Thr, Cys, Ser.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 51 1 is Ser, and the amino acid at or corresponding to position 544 is Thr. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 511 is Ser, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Gly, and the amino acid at or corresponding to position 511 is Ser, and the amino acid at or corresponding to position 544 is Ser. In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 511 is Ser, and the amino acid at or corresponding to position 544 is Thr.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 511 is Ser, and the amino acid at or corresponding to position 544 is Cys.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 463 is Cys, and the amino acid at or corresponding to position 511 is Ser, and the amino acid at or corresponding to position 544 is Ser.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 51 1 is Ser, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Val.

In another preferred embodiment, the mutated TK polypeptide comprises a sequence of SEQ ID NO: 1 a variant, derivative, orthologue, paralogue of homologue thereof, in which: the amino acid at or corresponding to position 51 1 is Ser, and the amino acid at or corresponding to position 514 is Asn, and the amino acid at or corresponding to position 544 is Val. It will be within the knowledge of the skilled artisan to identify conserved regions and motifs shared between the homologues, orthologues and paralogues encoded by SEQ ID NO: 182 or 183, such as those depicted in Table 1. Having identified such conserved regions that may represent suitable binding motifs, amino acids corresponding to the amino acids listed below in Table 2a and 2b, can be chosen to be subsituted by any other amino acid, for example by conserved amino acids, preferably by the amino acid substitutions described SUPRA using SEQ ID NO:1 as reference. Table 2a and 2b provides an overview of positions in the orthologues and homologues to SEQ ID NO:1 , i.e. the corresponding positions in SEQ ID NOs: 1 to 181 .

Table2a

ID Pos 1 Pos 2 Pos 3 Pos 4 Pos 5 Pos 6 Pos 7 Pos 8

1 I265 I267 Y337 S342 A343 L458 A459 S460

2 1195 1197 Y267 S272 A273 L388 A389 S390

3 I265 I267 Y337 S342 A343 L458 A459 S460

4 I276 I278 Y347 S352 A353 L468 A469 S470

5 I264 I266 Y336 S341 A342 L457 A458 S459

6 I263 I265 Y335 S340 A341 L456 A457 S458

7 I235 I237 H307 A312 P313 L428 A429 P430

8 I264 I266 Y336 A341 A342 L457 A458 S459

9 I266 I268 Y338 S343 A344 L459 A460 S461

10 I273 I275 Y345 S350 A351 L466 A467 S468

11 I247 I249 Y319 A324 A325 L440 A441 S442

12 I268 I270 Y340 S345 A346 L461 A462 S463

13 I272 I274 Y344 S349 A350 L465 A466 S467

14 I266 I268 Y338 S343 A344 L459 A460 S461

15 I268 I270 Y340 S345 A346 L461 A462 S463

16 I267 I269 Y339 A344 A345 L460 A461 S462

17 I264 I266 Y336 S341 A342 L457 A458 S459

18 I269 1271 Y341 S346 A347 L462 A463 S464

19 I263 I265 Y335 S340 A341 L456 A457 S458

20 1271 I273 Y343 S348 A349 L464 A465 S466

21 I276 I278 Y348 A353 A354 L469 A470 S471

22 I268 I270 Y340 S345 A346 L461 A462 S463

23 I270 I272 Y342 S347 A348 L463 A464 S465

24 I266 I268 Y338 S343 A344 L459 A460 S461

25 I259 1261 Y331 A336 A337 L452 A453 S454

26 I265 I267 Y337 A342 A343 L458 A459 S460

27 I268 I270 Y340 S345 A346 L461 A462 S463

28 I269 1271 Y341 S346 A347 L462 A463 S464

29 I270 I272 Y342 S347 A348 L463 A464 S465

30 I263 I265 Y335 A340 A341 L456 A457 S458

31 I268 I270 Y340 S345 A346 L461 A462 S463

32 I264 I267 Y337 A342 A343 L458 A459 S460

33 I269 1271 Y341 S346 A347 L462 A463 S464

34 I263 I265 Y335 A340 A341 L456 A457 S458

35 I264 I267 Y337 A342 A343 L458 A459 S460 I253 I255 Y325 S330 A331 L446 A447 S448

I265 I267 Y337 S342 A343 L458 A459 S460

I200 I202 Y272 S277 A278 L393 A394 S395

I264 I266 Y336 A341 A342 L457 A458 S459

I268 I270 Y340 S345 A346 L461 A462 S463

I265 I267 Y337 S342 G343 L458 A459 S460

I255 I257 Y327 S332 A333 L448 A449 S450

I265 I267 Y337 A342 A343 L471 A472 S473

I265 I267 Y337 A342 A343 L458 A459 S460

I268 I270 Y340 S345 A346 L461 A462 S463

I254 I256 Y326 S331 A332 L447 A448 S449

I203 I205 Y275 S280 A281 L396 A397 S398

I260 I262 Y332 A337 A338 L453 A454 S455

1199 1201 Y271 S276 A277 L392 A393 S394

I276 I278 Y348 S353 A354 L469 A470 S471

I225 I227 Y297 S302 A303 L418 A419 S420

I259 1261 Y331 S336 A337 L452 A453 S454

I266 I268 Y338 A343 A344 L459 A460 S461

I259 1261 Y331 S336 A337 L452 A453 S454

I269 1271 Y341 S346 A347 L462 A463 S464

I258 I260 Y330 S335 A336 L451 A452 S453

1158 1160 Y230 S235 A236 L351 A352 S353

I204 I206 Y276 A281 A282 L397 A398 S399

I274 I276 Y346 S351 A352 L467 A468 S469

1158 1160 Y230 A235 A236 L351 A352 S353

1158 1160 Y230 S235 A236 L351 A352 S353

1153 1155 Y225 S230 A231 L346 A347 S348

I260 I262 Y332 A337 A338 L453 A454 S455

1144 1146 Y216 A221 A222 L337 A338 S339

1158 1160 Y230 S235 A236 L351 A352 S353

I239 1155 Y225 S230 A231 L346 A347 S348

I236 I238 Y308 S313 A314 L429 A430 T431

I244 V31 1 Y381 G386 P387 L502 A503 P504

1215 I240 Y310 A315 A316 L431 A432 S433

I244 I246 Y316 S321 A322 L437 A438 T439

1215 1217 Y287 A292 A293 L408 A409 S410

I239 1241 Y31 1 S316 A317 L432 A433 T434 76 I239 1241 Y31 1 S316 A317 L432 A433 T434

77 1158 1160 Y230 A235 A236 L351 A352 S353

78 I246 I248 H318 S323 A324 L439 A440 S441

79 I239 1241 Y31 1 S316 A317 L432 A433 T434

80 I343 V345 Y415 G420 P421 L536 A537 P538

81 1251 I253 Y323 T328 A329 L444 A445 T446

82 I299 V301 Y371 G376 P377 L492 A493 P494

83 I232 I234 H304 S309 A310 L425 A426 S427

84 1153 1155 Y225 A230 A231 L346 A347 S348

85 1198 I200 Y270 N275 A276 L391 T392 L393

86 I253 I255 Y325 S330 A331 L446 A447 S448

87 1158 1160 Y230 S235 A236 L351 A352 S353

88 1146 1148 Y218 S223 A224 L339 A340 S341

89 1158 1160 Y230 N235 A236 L351 A352 T353

90 1158 1160 Y230 A235 A236 L351 A352 S353

91 11 14 11 16 Y186 S191 A192 L307 A308 S309

92 I94 I96 Y166 A171 A172 L287 A288 S289

93 1105 1107 Y177 S182 A183 L298 A299 S300

94 1123 V125 Y195 G200 A201 V316 A317 S318

95 I240 I242 H312 A317 P318 L433 A434 P435

96 I240 I242 H312 A317 P318 L433 A434 P435

97 1193 1195 A265 A270 A271 L386 T387 H388

98 I43 I45 Y1 15 S120 A121 L236 A237 S238

99 1193 1195 A265 A270 A271 L386 T387 H388

100 1193 1195 H265 A270 A271 L386 A387 P388

101 1193 1195 A265 A270 A271 L386 T387 H388

102 1193 1195 A265 A270 A271 L386 A387 K388

103 1194 1196 A266 A271 A272 L387 T388 H389

104 1190 1192 G261 N266 P267 L383 T384 G385

105 1188 1190 G259 A264 P265 L381 T382 G383

106 V192 1194 A263 S268 P269 L385 T386 G387

107 1193 1195 H264 S269 P270 L386 T387 G388

108 1185 1187 H255 S260 P261 L377 T378 G379

109 - - A264 G269 T270 L381 W382 N383

110 V199 A201 H270 N275 S276 L391 S392 V393

111 I205 C207 A275 A280 D281 L396 S397 P398

112 V214 V216 G285 A290 A291 L408 M409 E410

113 1190 1192 E261 S266 P267 L383 T384 G385

114 V219 V221 G290 A295 A296 L413 C414 E415

115 1188 1190 A259 A264 P265 L381 T382 P383 116 I227 I229 H298 N303 P304 L420 T421 G422

117 1190 1192 Y261 S266 P267 L384 T385 P386

118 I206 C208 A277 A282 A283 L400 S401 P402

119 1189 1191 A260 S265 A266 L383 T384 P385

120 1189 1191 E260 A265 P266 L382 T383 G384

121 1217 C219 A288 V293 A294 L41 1 S412 P413

122 V187 1189 A258 S263 P264 L380 T381 P382

123 I225 C227 A295 A300 A301 L418 T419 P420

124 1191 1193 H262 A267 P268 L384 T385 G386

125 1190 1192 C260 A265 P266 L382 T383 A384

126 V247 C249 F318 G323 A324 L441 V442 N443

127 V215 V217 G286 Q291 A292 L410 L41 1 E412

128 1189 C191 A260 A265 A266 L385 S386 P387

129 1201 C203 A272 A277 A278 L393 S394 P395

130 I209 C21 1 A280 V285 A286 L401 S402 P403

131 V199 C201 A270 G275 A276 L394 V395 T396

132 1191 C193 A261 A266 A267 L378 S379 P380

133 1188 1190 G259 A264 P265 L381 T382 G383

134 V192 1194 A263 S268 P269 L385 T386 G387

135 1193 1195 H264 S269 P270 L386 T387 G388

136 1190 1192 G261 N266 P267 L383 T384 S385

137 I225 C227 A296 S301 L302 L419 V420 N421

138 1185 1187 H255 S260 P261 L377 T378 G379

139 - - A264 G269 T270 L381 W382 N383

140 V199 A201 H270 N275 S276 L391 S392 V393

141 I205 C207 A275 A280 D281 L396 S397 P398

142 V214 V216 G285 A290 A291 L408 M409 E410

143 1193 1195 A264 A269 P270 L386 T387 G388

144 V200 C202 H271 S276 A277 L392 S393 V394

145 1190 1192 E261 S266 P267 L383 T384 G385

146 V219 V221 G290 A295 A296 L413 C414 E415

147 I226 I228 H297 N302 P303 L419 T420 P421

148 1192 C194 A263 A268 D269 L386 T387 P388

149 V224 C226 A295 A300 A301 L418 S419 P420

150 1199 C201 A270 A275 A276 L391 S392 P393

151 1188 1190 A259 A264 P265 L381 T382 P383

152 I227 I229 H298 N303 P304 L420 T421 G422

153 1190 1192 H260 S265 P266 L383 T384 P385

154 1190 1192 Y261 S266 P267 L384 T385 P386

155 V238 C240 A309 G314 G315 - - - 156 V199 1201 E263 E268 T269 L350 A351 G352

157 V201 I203 P265 S270 T271 L351 E352 G353

158 V201 I203 D265 T270 P271 L351 E352 G353

159 V202 I204 Q266 S271 N272 L353 E354 G355

160 V221 I223 E253 C258 N259 L272 K273 G274

161 V203 I205 D267 S272 T273 L354 E355 G356

162 V202 I204 K266 T271 T272 L352 E353 G354

163 V201 I203 E265 E270 S271 L352 E353 G354

164 V204 I206 E268 E273 S274 L355 E356 G357

165 V165 1167 K229 S234 T235 L315 A316 G317

166 V200 I202 A264 T269 P270 L352 E353 G354

167 V237 I239 K301 S306 P307 L386 E387 G388

168 V199 1201 E263 T268 C269 L354 E355 G356

169 V201 I203 E265 S270 E271 L351 E352 G353

170 L182 Q184 G253 K258 A259 V338 S339 N340

171 V205 I207 E269 S274 T275 L355 E356 G357

172 1197 1199 P266 K271 A272 L359 A360 G361

173 1196 1198 P265 K270 A271 L361 A362 G363

174 1185 1187 P254 K259 A260 L350 A351 G352

175 1195 1197 P264 K269 A270 L362 A363 S364

176 L185 Q187 D256 K261 P262 V341 S342 N343

177 V197 S199 S262 P267 L268 L375 A376 A377

178 1195 S197 S260 T265 A266 L353 M354 L355

179 1198 S200 A263 A268 A269 L358 K359 K360

180 V197 1199 1261 S266 T267 L347 E348 G349

181 1194 1196 P263 K268 P269 L360 A361 G362

Table 2b

ID Pos 9 Pos 10 Pos 11 Pos 12 Pos 13 Pos 14 Pos 15 Pos 16

1 S461 M463 T51 1 F512 F513 V514 F515 L544

2 S391 M393 T441 F442 F443 V444 F445 L474

3 S461 M463 T51 1 F512 F513 V514 F515 L544

4 S471 M473 T521 F522 F523 V524 F525 L554

5 S460 M462 T510 F51 1 F512 V513 F514 L543

6 S459 M461 T509 F510 F51 1 V512 F513 L542

7 S431 M433 T481 F482 F483 I484 F485 L514

8 S460 M462 T510 F51 1 F512 V513 F514 L543

9 S462 M464 T512 F513 F514 V515 F516 L545

10 S469 M471 T519 F520 F521 V522 F523 L552

11 S443 M445 T493 F494 F495 V496 F497 L526

12 S464 M466 T514 F515 F516 V517 F518 L547 S468 M470 T518 F519 F520 V521 F522 L551

S462 M464 T512 F513 F514 V515 F516 L545

S464 M466 T514 F515 F516 V517 F518 L547

S463 M465 T513 F514 F515 V516 F517 L546

S460 M462 T510 F51 1 F512 V513 F514 L543

S465 M467 T515 F516 F517 V518 F519 L548

S459 M461 T509 F510 F51 1 V512 F513 L542

S467 M469 T517 F518 F519 V520 F521 L550

S472 M474 T522 F523 F524 V525 F526 L555

S464 M466 T514 F515 F516 V517 F518 L547

S466 M468 T516 F517 F518 V519 F520 L549

S462 M464 T512 F513 F514 V515 F516 L545

S455 M457 T505 F506 F507 V508 F509 L538

S461 M463 T51 1 F512 F513 V514 F515 L544

S464 M466 T514 F515 F516 V517 F518 L547

S465 M467 T515 F516 F517 V518 F519 L548

S466 M468 T516 F517 F518 V519 F520 L549

S459 M461 T509 F510 F51 1 V512 F513 L542

S464 M466 T514 F515 F516 V517 F518 L547

S461 M463 T51 1 F512 F513 V514 F515 L544

S465 M467 T515 F516 F517 V518 F519 L548

S459 M461 T509 F510 F51 1 V512 F513 L542

S461 M463 T51 1 F512 F513 V514 F515 L544

S449 M451 T499 F500 F501 V502 F503 L532

S461 M463 T51 1 F512 F513 V514 F515 L544

S396 M398 T446 F447 F448 V449 F450 L479

S460 M462 T510 F51 1 F512 V513 F514 L543

S464 M466 T514 F515 F516 V517 F518 L547

S461 M463 T51 1 F512 F513 V514 F515 L544

S451 M453 T501 F502 F503 V504 F505 L534

S474 M476 T524 F525 F526 V527 F528 L557

S461 M463 T51 1 F512 F513 V514 F515 L544

S464 M466 T514 F515 F516 V517 F518 L547

S450 M452 T500 F501 F502 V503 F504 L533

S399 M401 T449 F450 F451 V452 F453 L482

S456 M458 T506 F507 F508 V509 F510 L539

S395 M397 T445 F446 F447 V448 F449 L478

S472 M474 T521 F522 F523 V524 F525 L554

S421 M423 T471 F472 F473 V474 F475 L504

S455 M457 T505 F506 F507 V508 F509 L538 S462 M464 T512 F513 F514 V515 F516 L545

S455 M457 T505 F506 F507 V508 F509 L538

S465 L467 T515 F516 F517 V518 F519 L548

S454 M456 T504 F505 F506 V507 F508 L537

S354 M356 T404 F405 F406 V407 F408 L437

S400 M402 T450 F451 F452 V453 F454 L483

S470 M472 T520 F521 F522 V523 F524 L553

S354 M356 T404 F405 F406 V407 F408 L437

S349 M351 T399 F400 F401 V402 F403 L432

S456 M458 T506 F507 F508 V509 F510 L539

S340 M342 T390 F391 F392 V393 F394 L423

S354 M356 T404 F405 F406 V407 F408 L437

S349 M351 T399 F400 F401 V402 F403 L432

S432 M434 T482 F483 F484 V485 F486 L515

S505 M507 T555 Y556 F557 A558 F559 V588

S434 M436 T484 F485 F486 I487 F488 L517

S440 M442 T490 F491 F492 V493 F494 L523

S41 1 M413 T461 F462 F463 I464 F465 L494

S435 M437 T485 F486 F487 V488 F489 L518

S354 M356 T404 F405 F406 V407 F408 L437

S442 M444 T492 F493 F494 V495 F496 L525

S435 M437 T485 F486 F487 V488 F489 L518

S539 M541 T589 Y590 F591 A592 F593 V622

S447 M449 T497 F498 F499 V500 F501 L530

S495 M497 T545 Y546 F547 A548 F549 V578

S428 M430 T478 F479 F480 V481 F482 L51 1

S349 M351 T399 F400 F401 V402 F403 L432

S394 M396 T444 Y445 F446 V447 F448 L477

S449 M451 T499 F500 F501 V502 F503 -

S354 M356 T404 F405 F406 V407 F408 L437

S342 M344 T392 F393 F394 V395 F396 L425

S354 M356 T404 F405 F406 V407 F408 L437

S354 M356 T404 F405 F406 I407 F408 L437

S310 M312 T360 F361 F362 V363 F364 L393

S290 M292 T340 F341 F342 V343 F344 L373 93 S301 M303 T351 F352 F353 V354 F355 L384

94 S319 M321 T369 Y370 L371 A372 F373 V402

95 S436 L438 T486 F487 Y488 I489 F490 L519

96 S436 L438 T486 F487 Y488 I489 F490 L519

97 S389 L391 T439 F440 L441 V442 F443 L472

98 S239 M241 T289 F290 F291 V292 F293 L322

99 S389 N391 T439 F440 L441 V442 F443 L472

100 S389 L391 T439 F440 L441 I442 F443 L472

101 S389 L391 T439 F440 L441 V442 F443 L472

102 S389 N391 T439 F440 L441 V442 F443 L472

103 S390 Y392 T440 F441 L442 V443 F444 L473

104 S386 N388 T440 F441 L442 N443 F444 L473

105 S384 L386 T439 F440 L441 N442 F443 L472

106 S388 L390 T442 F443 L444 N445 F446 L475

107 S389 L391 T443 F444 L445 N446 F447 L476

108 S380 L382 T435 F436 L437 N438 F439 L468

109 S384 Q386 T435 F436 F437 M438 F439 E468

110 S394 N396 T455 F456 F457 M458 F459 E488

111 S399 N401 T460 F461 F462 M463 F464 T493

112 S41 1 F413 S467 Y468 L469 A470 F471 V500

113 S386 L388 T440 F441 F442 N443 F444 L473

114 S416 F418 S472 Y473 F474 I475 F476 V505

115 S384 L386 T439 F440 L441 N442 F443 L472

116 S423 L425 T479 F480 A481 N482 F483 L512

117 S387 L389 T442 F443 L444 N445 F446 L475

118 S403 H405 S464 F465 F466 M467 F468 M497

119 S386 L388 T441 F442 L443 N444 F445 L474

120 S385 L387 T439 F440 L441 N442 F443 L472

121 S414 N416 S475 F476 F477 M478 F479 T508

122 S383 N385 T438 F439 L440 N441 F442 L471

123 S421 H423 S482 F483 F484 M485 F486 T515

124 S387 L389 T441 F442 L443 N444 F445 L474

125 S385 L387 T439 F440 L441 N442 F443 L472

126 S444 K446 T496 F497 F498 M499 F500 E529

127 S413 F415 T469 F470 F471 M472 F473 I502

128 S388 N390 S449 F450 F451 M452 F453 T482

129 S396 N398 S457 F458 F459 M460 F461 T490

130 S404 N406 S465 F466 F467 M468 F469 M498

131 S397 K399 T451 F452 L453 M454 F455 E484

132 S381 N383 S442 F443 F444 M445 F446 L475 133 S384 L386 T439 F440 L441 N442 F443 L472

134 S388 L390 T442 F443 L444 N445 F446 L475

135 S389 L391 T443 F444 L445 N446 F447 L476

136 S386 N388 T440 F441 F442 N443 F444 L473

137 S422 K424 T474 F475 F476 M477 F478 E507

138 S380 L382 T435 F436 L437 N438 F439 L468

139 S384 Q386 T435 F436 F437 M438 F439 E468

140 S394 N396 T455 F456 F457 M458 F459 E488

141 S399 N401 T460 F461 F462 M463 F464 T493

142 S41 1 F413 S467 Y468 L469 A470 F471 V500

143 S389 L391 T443 F444 L445 N446 F447 L476

144 S395 L397 T456 F457 Y458 M459 F460 A489

145 S386 L388 T440 F441 F442 N443 F444 L473

146 S416 F418 S472 Y473 F474 I475 F476 V505

147 S422 L424 T478 F479 L480 N481 F482 L51 1

148 S389 N391 S450 F451 F452 M453 F454 T483

149 S421 N423 S482 F483 F484 M485 F486 T515

150 S394 N396 S455 F456 F457 M458 F459 I488

151 S384 L386 T439 F440 L441 N442 F443 L472

152 S423 L425 T479 F480 A481 N482 F483 L512

153 S386 L388 T441 F442 L443 N444 F445 V474

154 S387 L389 T442 F443 L444 N445 F446 L475

155 - - T434 F435 F436 M437 F438 E467

156 S353 G355 T398 F399 S400 A401 F402 E431

157 S354 G356 T397 F398 S399 A400 F401 E430

158 S354 G356 T397 F398 A399 A400 F401 E430

159 S356 G358 T399 F400 S401 A402 F403 E432

160 S275 G277 - - - - - -

161 S357 G359 T402 F403 T404 A405 F406 E435

162 S355 G357 T398 F399 S400 A401 F402 E431

163 S355 G357 T400 F401 S402 A403 F404 E433

164 S358 G360 T403 F404 S405 A406 F407 E436

165 S318 G320 T361 F362 S363 A364 F365 E394

166 S355 G357 T398 F399 G400 A401 F402 E431

167 S389 G391 T433 F434 S435 A436 F437 E466

168 S357 G359 T400 F401 A402 A403 F404 D433

169 S354 G356 T397 F398 S399 A400 F401 D430

170 S341 Y343 T383 F384 A385 A386 F387 1416

171 S358 G360 T401 F402 S403 A404 F405 D434

172 S362 K364 G404 F405 G406 M407 F408 V438 173 S364 K366 D406 F407 G408 V409 F410 V440

174 S353 K355 D395 F396 G397 V398 F399 V429

175 S365 K367 D407 F408 G409 V410 F41 1 V441

176 S344 Y346 T386 F387 A388 A389 F390 1419

177 D378 K380 S420 F421 A422 S423 F424 P454

178 D356 G358 S398 F399 A400 C401 F402 P432

179 D361 G363 S403 F404 A405 C406 F407 P437

180 S350 G352 T393 F394 S395 A396 F397 D426

181 S363 K365 D405 F406 G407 V408 F409 V439

a) generating a library of mutated TK-encoding nucleic acids,

d) selecting at least one mutated TK-encoding nucleic acid that provides a significantly

increased level of tolerance to a TK-inhibiting herbicide as compared to that provided by the control TK-encoding nucleic acid. In a preferred embodiment, the mutated TK-encoding nucleic acid selected in step d) provides at least 2-fold as much resistance or tolerance of a cell or plant to a TK-inhibiting herbicide as compared to that provided by the control TK-encoding nucleic acid.

In a further preferred embodiment, the mutated TK-encoding nucleic acid selected in step d) provides at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold, at least 500-fold, as much resistance or tolerance of a cell or plant to a TK-inhibiting herbicide as compared to that provided by the control TK-encoding nucleic acid.

The resistance or tolerance can be determined by generating a transgenic plant or host cell, preferably a plant cell, comprising a nucleic acid sequence of the library of step a) and comparing said transgenic plant with a control plant or host cell, preferably a plant cell.

Another object refers to a method of identifying a plant or algae containing a nucleic acid comprising a nucleotide sequence encoding a wild-type or mutated TK which is resistant or tolerant to a TK-inhibiting herbicide, the method comprising:

a) identifying an effective amount of a TK-inhibiting herbicide in a culture of plant cells or green algae that leads to death of said cells.

b) treating said plant cells or green algae with a mutagenizing agent,

d) selecting at least one cell surviving these test conditions,

In a preferred embodiment, said mutagenizing agent is ethylmethanesulfonate (EMS).

Many methods well known to the skilled artisan are available for obtaining suitable candidate nucleic acids for identifying a nucleotide sequence encoding a mutated TK from a variety of different potential source organisms including microbes, plants, fungi, algae, mixed cultures etc. as well as environmental sources of DNA such as soil. These methods include inter alia the preparation of cDNA or genomic DNA libraries, the use of suitably degenerate oligonucleotide primers, the use of probes based upon known sequences or complementation assays (for example, for growth upon tyrosine) as well as the use of mutagenesis and shuffling in order to provide recombined or shuffled mutated TK-encoding sequences.

Nucleic acids comprising candidate and control TK encoding sequences can be expressed in yeast, in a bacterial host strain, in an alga or in a higher plant such as tobacco or Arabidopsis and the relative levels of inherent tolerance of the TK encoding sequences screened according to a visible indicator phenotype of the transformed strain or plant in the presence of different concentrations of the selected TK-inhibiting herbicide. Dose responses and relative shifts in dose responses associated with these indicator phenotypes (formation of brown color, growth inhibition, herbicidal effect etc) are conveniently expressed in terms, for example, of GR50 (concentration for 50% reduction of growth) or MIC (minimum inhibitory concentration) values where increases in values correspond to increases in inherent tolerance of the expressed TK. For example, in a relatively rapid assay system based upon transformation of a bacterium such as E. coli, each mutated TK encoding sequence may be expressed, for example, as a DNA sequence under expression control of a controllable promoter such as the lacZ promoter and taking suitable account, for example by the use of synthetic DNA, of such issues as codon usage in order to obtain as comparable a level of expression as possible of different TK sequences. Such strains expressing nucleic acids comprising alternative candidate TK sequences may be plated out on different concentrations of the selected TK-inhibiting herbicide in, optionally, a tyrosine supplemented medium and the relative levels of inherent tolerance of the expressed TK enzymes estimated on the basis of the extent and MIC for inhibition of the formation of the brown, ochronotic pigment.

In another embodiment, candidate nucleic acids are transformed into plant material to generate a transgenic plant, regenerated into morphologically normal fertile plants which are then measured for differential tolerance to selected TK-inhibiting herbicides as described in the Example section hereinafter. Many suitable methods for transformation using suitable selection markers such as kanamycin, binary vectors such as from Agrobacterium and plant regeneration as, for example, from tobacco leaf discs are well known in the art. Optionally, a control population of plants is likewise transformed with a nucleic acid expressing the control TK. Alternatively, an untransformed dicot plant such as Arabidopsis or Tobacco can be used as a control since this, in any case, expresses its own endogenous TK. The average, and distribution, of herbicide tolerance levels of a range of primary plant transformation events or their progeny to TK-inhibiting herbicides described supra are evaluated in the normal manner based upon plant damage, meristematic bleaching symptoms etc. at a range of different concentrations of herbicides. These data can be expressed in terms of, for example, GR50 values derived from dose/response curves having "dose" plotted on the x-axis and "percentage kill", "herbicidal effect", "numbers of emerging green plants" etc. plotted on the y-axis where increased GR50 values correspond to increased levels of inherent tolerance of the expressed TK. Herbicides can suitably be applied pre-emergence or post-emergence.

Another object of the present invention refers to an isolated nucleic acid encoding a mutated TK as disclosed SUPRA, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 182, or 183, or a variant or derivative thereof.

In one embodiment, the nucleic acid is identifiable by a method as defined above.

In a preferred embodiment, the encoded mutated TK is a variant of SEQ ID NO: 1 , which includes one or more of the following:

the amino acid corresponding to position 265 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,Met,Phe,Tyr, or Trp;

the amino acid corresponding to position 267 of SEQ ID NO: 1 is

the amino acid corresponding to position 337 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Met,Phe, or Trp;

the amino acid corresponding to position 342 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Met,Phe,Tyr, or Trp;

the amino acid corresponding to position 343 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Val,Leu,lle,Met,Phe,Tyr, or Trp, the amino acid corresponding to position 458 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,lle,Met,Phe,Tyr, or Trp, the amino acid corresponding to position 459 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Val,Leu,lle,Met,Phe,Tyr, or Trp the amino acid corresponding to position 460 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Met,Phe,Tyr, or Trp, the amino acid corresponding to position 461 of SEQ ID NO:1 is

Arg,His,Lys,Asp,Glu,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Met,Phe,Tyr, or Trp.

the amino acid corresponding to position 463 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Phe,Tyr, or Trp.

the amino acid corresponding to position 51 1 of SEQ ID NO: 1 is

Arg,His,Lys,Asp,Glu,Ser,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Phe,Met,Tyr, or Trp.

the amino acid corresponding to position 512 of SEQ ID NO: 1 is Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Met,Tyr, or Trp.

the amino acid corresponding to position 513 of SEQ ID NO:1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,Leu,lle,Met,Tyr, or Trp.

the amino acid corresponding to position 514 of SEQ ID NO:1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Leu,lle,Phe,Met,Tyr, or Trp.

the amino acid corresponding to position 515 of SEQ ID NO:1 is

the amino acid corresponding to position 544 of SEQ ID NO:1 is

Arg,His,Lys,Asp,Glu,Ser,Thr,Asn,Gln,Cys,Gly,Pro,Ala,Val,lle,Met,Phe,Tyr, or Trp

In other aspects, the present invention encompasses a progeny or a descendant of a TK- inhibiting herbicides-tolerant plant of the present invention as well as seeds derived from the TK-inhibiting herbicides-tolerant plants of the invention and cells derived from the TK- inhibiting herbicides-tolerant plants of the invention.

In some embodiments, the present invention provides a progeny or descendant plant derived from a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, wherein the progeny or descendant plant comprises in at least some of its cells the recombinant polynucleotide operably linked to the promoter, the expression of the wildtype or mutated TK polypeptide conferring to the progeny or descendant plant tolerance to the TK-inhibiting herbicides.

In one embodiment, seeds of the present invention preferably comprise the TK-inhibiting herbicides-tolerance characteristics of the TK-inhibiting herbicides-tolerant plant. In other embodiments, a seed is capable of germination into a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the progeny or descendant plant tolerance to the TK-inhibiting herbicides.

In some embodiments, plant cells of the present invention are capable of regenerating a plant or plant part. In other embodiments, plant cells are not capable of regenerating a plant or plant part. Examples of cells not capable of regenerating a plant include, but are not limited to, endosperm, seed coat (testa & pericarp), and root cap.

In another embodiment, the present invention provides a plant cell of or capable of regenerating a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to the TK-inhibiting herbicides, wherein the plant cell comprises the recombinant polynucleotide operably linked to a promoter.

In other embodiments, the present invention provides a plant cell comprising a

polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the cell tolerance to the TK-inhibiting herbicides.

In another embodiment, the invention refers to a plant cell transformed by a nucleic acid encoding a mutated TK polypeptide according to the present invention or to a plant cell which has been mutated to obtain a plant expressing a nucleic acid encoding a mutated TK polypeptide according to the present invention, wherein expression of the nucleic acid in the plant cell results in increased resistance or tolerance to a TK-inhibiting herbicide as compared to a wild type variety of the plant cell. Preferably, the mutated TK polypeptide encoding nucleic acid comprises a polynucleotide sequence selected from the group consisting of: a) a polynucleotide as shown in SEQ ID NO: 182, or 183, or a variant or derivative thereof; b) a polynucleotide encoding a polypeptide as shown in SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 1 11 , 112, 1 13, 1 14, 1 15, 1 16, 117, 1 18, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , or a variant or derivative thereof; c) a polynucleotide comprising at least 60 consecutive nucleotides of any of a) or b); and d) a polynucleotide complementary to the polynucleotide of any of a) through c). In some aspects, the present invention provides a plant product prepared from the TK- inhibiting herbicides-tolerant plants hereof. In some embodiments, examples of plant products include, without limitation, grain, oil, and meal. In one embodiment, a plant product is plant grain (e.g., grain suitable for use as feed or for processing), plant oil (e.g., oil suitable for use as food or biodiesel), or plant meal (e.g., meal suitable for use as feed).

In one embodiment, a plant product prepared from a plant or plant part is provided, wherein the plant or plant part comprises in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the a plant or plant part tolerance to the TK-inhibiting herbicides. In another embodiment, the invention refers to a method of producing a transgenic plant cell with an increased resistance to a TK-inhibiting herbicide as compared to a wild type variety of the plant cell comprising, transforming the plant cell with an expression cassette comprising a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide.

In another embodiment, the invention refers to a method of producing a transgenic plant comprising, (a) transforming a plant cell with an expression cassette comprising a a

polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, and (b) generating a plant with an increased resistance to TK-inhibiting herbicide from the plant cell.

In some aspects, the present invention provides a method for producing a TK-inhibiting herbicides-tolerant plant. In one embodiment, the method comprises: regenerating a plant from a plant cell transformed with a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to the TK-inhibiting herbicides. The term "expression/expressing" or "gene expression" means the transcription of a specific gene or specific genes or specific genetic construct. The term "expression" or "gene expression" in particular means the transcription of a gene or genes or genetic construct into structural RNA (rRNA, tRNA) or mRNA with or without subsequent translation of the latter into a protein. The process includes transcription of DNA and processing of the resulting mRNA product.

To obtain the desired effect, i.e. plants that are tolerant or resistant to the TK-inhibiting herbicide derivative herbicide of the present invention, it will be understood that the at least one nucleic acid is "over-expressed" by methods and means known to the person skilled in the art.

The term "increased expression" or "overexpression" as used herein means any form of expression that is additional to the original wild-type expression level. Methods for increasing expression of genes or gene products are well documented in the art and include, for example, overexpression driven by appropriate promoters, the use of transcription enhancers or translation enhancers. Isolated nucleic acids which serve as promoter or enhancer elements may be introduced in an appropriate position (typically upstream) of a non-heterologous form of a polynucleotide so as to upregulate expression of a nucleic acid encoding the polypeptide of interest. For example, endogenous promoters may be altered in vivo by mutation, deletion, and/or substitution (see, Kmiec, US 5,565,350; Zarling et al., W09322443), or isolated promoters may be introduced into a plant cell in the proper orientation and distance from a gene of the present invention so as to control the expression of the gene. If polypeptide expression is desired, it is generally desirable to include a polyadenylation region at the 3'-end of a polynucleotide coding region. The polyadenylation region can be derived from the natural gene, from a variety of other plant genes, or from T-DNA. The 3' end sequence to be added may be derived from, for example, the nopaline synthase or octopine synthase genes, or alternatively from another plant gene, or less preferably from any other eukaryotic gene. An intron sequence may also be added to the 5' untranslated region (UTR) or the coding sequence of the partial coding sequence to increase the amount of the mature message that accumulates in the cytosol. Inclusion of a spliceable intron in the transcription unit in both plant and animal expression constructs has been shown to increase gene expression at both the mRNA and protein levels up to 1000-fold (Buchman and Berg (1988) Mol. Cell biol. 8: 4395-4405; Callis et al. (1987) Genes Dev 1 :1183- 1200). Such intron enhancement of gene expression is typically greatest when placed near the 5' end of the transcription unit. Use of the maize introns Adh1-S intron 1 , 2, and 6, the Bronze-1 intron are known in the art. For general information see: The Maize Handbook, Chapter 1 16, Freeling and Walbot, Eds., Springer, N.Y. (1994).

Where appropriate, nucleic acid sequences may be optimized for increased expression in a transformed plant. For example, coding sequences that comprise plant-preferred codons for improved expression in a plant can be provided. See, for example, Campbell and Gowri (1990) Plant Physiol., 92: 1 -11 for a discussion of host-preferred codon usage. Methods also are known in the art for preparing plant-preferred genes. See, for example, U.S. Patent Nos. 5,380,831 , and 5,436,391 , and Murray et al. (1989) Nucleic Acids Res. 17:477-498, herein incorporated by reference. Consequently, wildtype/mutated TK nucleic acids of the invention are provided in

expression cassettes for expression in the plant of interest. The cassette will include regulatory sequences operably linked to a mutated TK nucleic acid sequence of the invention. The term "regulatory element" as used herein refers to a polynucleotide that is capable of regulating the transcription of an operably linked polynucleotide. It includes, but not limited to, promoters, enhancers, introns, 5' UTRs, and 3' UTRs. By "operably linked" is intended a functional linkage between a promoter and a second sequence, wherein the promoter sequence initiates and mediates transcription of the DNA sequence

corresponding to the second sequence. Generally, operably linked means that the nucleic acid sequences being linked are contiguous and, where necessary to join two protein coding regions, contiguous and in the same reading frame. The cassette may additionally contain at least one additional gene to be cotransformed into the organism. Alternatively, the additional gene(s) can be provided on multiple expression cassettes.

Such an expression cassette is provided with a plurality of restriction sites for insertion of the wildtype/mutated TK nucleic acid sequence to be under the transcriptional regulation of the regulatory regions. The expression cassette may additionally contain selectable marker genes. The expression cassette of the present invention will include in the 5'-3' direction of transcription, a transcriptional and translational initiation region (i.e., a promoter), a wildtype/mutated TK encoding nucleic acid sequence of the invention, and a transcriptional and translational termination region (i.e., termination region) functional in plants. The promoter may be native or analogous, or foreign or heterologous, to the plant host and/or to the wildtype/mutated TK nucleic acid sequence of the invention. Additionally, the promoter may be the natural sequence or alternatively a synthetic sequence. Where the promoter is "foreign" or "heterologous" to the plant host, it is intended that the promoter is not found in the native plant into which the promoter is introduced. Where the promoter is "foreign" or "heterologous" to the wildtype/mutated TK nucleic acid sequence of the invention, it is intended that the promoter is not the native or naturally occurring promoter for the operably linked wildtype/mutated TK nucleic acid sequence of the invention. As used herein, a chimeric gene comprises a coding sequence operably linked to a transcription initiation region that is heterologous to the coding sequence. While it may be preferable to express the wildtype/mutated TK nucleic acids of the invention using heterologous promoters, the native promoter sequences may be used. Such constructs would change expression levels of the wildtype/mutated TK protein in the plant or plant cell. Thus, the phenotype of the plant or plant cell is altered.

The termination region may be native with the transcriptional initiation region, may be native with the operably linked wildtype/mutated TK sequence of interest, may be native with the plant host, or may be derived from another source (i.e., foreign or heterologous to the promoter, the wildtype/mutated TK nucleic acid sequence of interest, the plant host, or any combination thereof). Convenient termination regions are available from the Ti-plasmid of A. tumefaciens , such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al. (1991 ) Mol. Gen. Genet. 262: 141 -144; Proudfoot (1991) Cell 64:671-674; Sanfacon et al. (1991 ) Genes Dev. 5: 141 -149; Mogen et al. (1990) Plant Cell 2: 1261 -1272; Munroe et al. (1990) Gene 91 : 151 -158; Ballas t al. (1989) Nucleic Acids Res. 17:7891 -7903; and Joshi et al. (1987) Nucleic Acid Res. 15:9627-9639. Where appropriate, the gene(s) may be optimized for increased expression in the transformed plant. That is, the genes can be synthesized using plant-preferred codons for improved expression. See, for example, Campbell and Gowri (1990) Plant Physiol. 92: 1 -1 1 for a discussion of host-preferred codon usage. Methods are available in the art for synthesizing plant-preferred genes. See, for example, U.S. Patent Nos. 5,380,831 , and 5,436,391 , and Murray et al. (1989) Nucleic Acids Res. 17:477-498, herein incorporated by reference.

While the polynucleotides of the invention may find use as selectable marker genes for plant transformation, the expression cassettes of the invention can include another selectable marker gene for the selection of transformed cells. Selectable marker genes, including those of the present invention, are utilized for the selection of transformed cells or tissues. Marker genes include, but are not limited to, genes encoding antibiotic resistance, such as those encoding neomycin phosphotransferase II (NEO) and hygromycin

phosphotransferase (HPT), as well as genes conferring resistance to herbicidal compounds, such as glufosinate ammonium, bromoxynil, imidazolinones, and 2,4- dichlorophenoxyacetate (2,4-D). See generally, Yarranton (1992) Curr. Opin. Biotech. 3 :506-511 ; Christophers on et al (1992) Proc. Natl. Acad. ScL USA 89:6314-6318; Yao et al. (1992) Cell 71 :63-72; Reznikoff (1992) Mol Microbiol 6:2419-2422; Barkley et al (1980) in The Operon, pp. 177-220; Hu et al (1987) Cell 48:555-566; Brown et al (1987) Cell 49:603- 612; Figge et al (1988) Cell 52:713-722; Deuschle et al (1989) Proc. Natl Acad. AcL USA 86:5400-5404; Fuerst et al (1989) Proc. Natl Acad. ScL USA 86:2549-2553; Deuschle et al

(1990) Science 248:480-483; Gossen (1993) Ph.D. Thesis, University of Heidelberg; Reines et al (1993) Proc. Natl Acad. ScL USA 90: 1917-1921 ; Labow et al (1990) Mol Cell Biol 10:3343-3356; Zambretti et al (1992) Proc. Natl Acad. ScL USA 89:3952-3956; Bairn et al

(1991 ) Proc. Natl Acad. ScL USA 88:5072-5076; Wyborski et al (1991) Nucleic Acids Res. 19:4647-4653; Hillenand-Wissman (1989) Topics Mol Struc. Biol 10: 143- 162; Degenkolb et al (1991 ) Antimicrob. Agents Chemother. 35: 1591 -1595; Kleinschnidt et al (1988) Biochemistry 27: 1094-1 104; Bonin (1993) Ph.D. Thesis, University of Heidelberg; Gossen et al (1992) Proc. Natl Acad. ScL USA 89:5547- 5551 ; Oliva et al (1992) Antimicrob. Agents Chemother. 36:913-919; Hlavka et al (1985) Handbook of Experimental Pharmacology, Vol. 78 ( Springer- Verlag, Berlin); Gill et al (1988) Nature 334:721 -724. Such disclosures are herein incorporated by reference. The above list of selectable marker genes is not meant to be limiting. Any selectable marker gene can be used in the present invention.

Further, additional sequence modifications are known to enhance gene expression in a cellular host. These include elimination of sequences encoding spurious polyadenylation signals, exon-intron splice site signals, transposon-like repeats, and other such well - characterized sequences that may be deleterious to gene expression. The G-C content of the sequence may be adjusted to levels average for a given cellular host, as calculated by reference to known genes expressed in the host cell. Also, if desired, sequences can be readily modified to avoid predicted hairpin secondary mRNA structures. Nucleotide sequences for enhancing gene expression can also be used in the plant expression vectors. These include, for example, introns of the maize Adh gene Adh1 -S intron 1 , 2, and 6 (Callis et al. Genes and Development 1 : 1 183-1200, 1987), and leader sequences, (W- sequence) from the Tobacco Mosaic virus (TMV), Maize Chlorotic Mottle Virus and Alfalfa Mosaic Virus (Gallie et al. Nucleic Acid Res. 15:8693-871 1 , 1987 and Skuzeski et al. Plant Mol. Biol. 15:65-79, 1990). The first intron from the shrunken-1 locus of maize has been shown to increase expression of genes in chimeric gene constructs. U.S. Pat. Nos.

5,424,412 and 5,593,874 disclose the use of specific introns in gene expression constructs, and Gallie et al. (Plant Physiol. 106:929-939, 1994) also have shown that introns are useful for regulating gene expression on a tissue specific basis. To further enhance or to optimize gene expression, the plant expression vectors of the invention also may contain DNA sequences containing matrix attachment regions (MARs). Plant cells transformed with such modified expression systems, then, may exhibit overexpression or constitutive expression of a nucleotide sequence of the invention. The invention further provides an isolated recombinant expression vector comprising the expression cassette containing a wildtype/mutated TK nucleic acid nucleic acid as described above, wherein expression of the vector in a host cell results in increased tolerance to a TK-inhibiting herbicide as compared to a wild type variety of the host cell. As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid," which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal

mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "expression vectors." In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, "plasmid" and "vector" can be used interchangeably as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses), which serve equivalent functions.

The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operably linked to the nucleic acid sequence to be expressed. Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cells and those that direct expression of the nucleotide sequence only in certain host cells or under certain conditions. It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of polypeptide desired, etc. The expression vectors of the invention can be introduced into host cells to thereby produce polypeptides or peptides, including fusion polypeptides or peptides, encoded by nucleic acids as described herein (e.g., mutated TK polypeptides, fusion polypeptides, etc.)

Expression vectors may additionally contain 5' leader sequences in the expression construct. Such leader sequences can act to enhance translation. Translation leaders are known in the art and include: picornavirus leaders, for example, EMCV leader

(Encephalomyo carditis 5' noncoding region) (Elroy-Stein et al. (1989) PNAS, 86:6126-

6130); potyvirus leaders, for example, TEV leader (Tobacco Etch Virus) (Gallie et al. (1995) Gene 165(2):233-238), MDMV leader (Maize Dwarf Mosaic Virus) (Virology 154:9-20), and human immunoglobulin heavy-chain binding protein (BiP) (Macejak et al. (1991 ) Nature 353:90-94); untranslated leader from the coat protein mRNA of alfalfa mosaic virus (AMV RNA 4) (Jobling et al. (1987) Nature 325:622-625); tobacco mosaic virus leader (TMV) (Gallie et al. (1989) in Molecular Biology of RNA, ed. Cech (Liss, New York), pp. 237-256); and maize chlorotic mottle virus leader (MCMV) (Lommel et al. (1991) Virology 81 :382- 385). See also, Della-Cioppa et al. (1987) Plant Physiol. 84:965-968.

Other methods known to enhance translation also can be utilized, for example, introns, and the like. In preparing an expression vector, the various nucleic acid fragments may be manipulated, so as to provide for the nucleic acid sequences in the proper orientation and, as appropriate, in the proper reading frame. Toward this end, adapters or linkers may be employed to join the nucleic acid fragments or other manipulations may be involved to provide for convenient restriction sites, removal of superfluous nucleic acid, removal of restriction sites, or the like. For this purpose, in vitro mutagenesis, primer repair, restriction, annealing, resubstitutions, e.g., transitions and transversions, may be involved.

A number of promoters can be used in the practice of the invention. The promoters can be selected based on the desired outcome. The nucleic acids can be combined with

constitutive, tissue-preferred, or other promoters for expression in plants.

Constitutive promoters include, for example, the core promoter of the Rsyn7 promoter and other constitutive promoters disclosed in WO 99/43838 and U.S. Patent No. 6,072,050; the core CaMV 35S promoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroy et al. (1990) Plant Cell 2: 163-171 ); ubiquitin (Christensen et al. (1989) Plant Mol. Biol. 12:619- 632 and Christensen et al. (1992) Plant Mol. Biol. 18:675-689); pEMU (Last et al. (1991 ) Theor. Appl. Genet. 81 :581 - 588); MAS (Velten et al. (1984) EMBO J. 3:2723- 2730); ALS promoter (U.S. Patent No. 5,659,026), and the like. Other constitutive promoters include, for example, U.S. Patent Nos. 5,608,149; 5,608, 144; 5,604,121 ; 5,569,597; 5,466,785;

5,399,680; 5,268,463; 5,608,142; and 6,177,611.

Tissue-preferred promoters can be utilized to target enhanced expression within a particular plant tissue. Such tissue-preferred promoters include, but are not limited to, leaf- preferred promoters, root-preferred promoters, seed- preferred promoters, and stem-preferred promoters. Some examples of tissue-preferred promoters are described by, e.g.,

Yamamoto et al. (1997) Plant J. 12(2):255-265; Kawamata et al. (1997) Plant Cell Physiol. 38(7):792-803; Hansen et al. (1997) Mol. Gen Genet. 254(3):337-343; Russell et al. (1997) Transgenic Res. 6(2): 157-168; Rinehart et al. (1996) Plant Physiol. 1 12(3): 1331 -1341 ; Van Camp et al. (1996) Plant Physiol. 1 12(2):525-535; Canevascini et al. (1996) Plant Physiol. 1 12(2):513- 524; Yamamoto et al. (1994) Plant Cell Physiol. 35(5):773-778; Lam (1994) Results Probl. Cell Differ. 20:181 - 196; Orozco ef al. (1993) Plant Mol Biol. 23(6): 1 129-1 138; Matsuoka et al. (1993) Voc Natl. Acad. ScL USA 90(20):9586-9590; and

Guevara-Garcia et al. (1993) Plant J 4(3):495-505. Promoters can be modified, if necessary, for weak expression.

In some embodiments, the nucleic acids of interest can be targeted to the chloroplast for expression. In this manner, where the nucleic acid of interest is not directly inserted into the chloroplast, the expression vector will additionally contain a chloroplast- targeting sequence comprising a nucleotide sequence that encodes a chloroplast transit peptide to direct the gene product of interest to the chloroplasts. Such transit peptides are known in the art. With respect to chloroplast-targeting sequences, "operably linked" means that the nucleic acid sequence encoding a transit peptide (i.e., the chloroplast-targeting sequence) is linked to the desired coding sequence of the invention such that the two sequences are contiguous and in the same reading frame. See, for example, Von Heijne et al. (1991 ) Plant Mol. Biol. Rep. 9: 104-126; Clark et al. (1989) J Biol. Chem. 264:17544-17550; Della-Cioppa et al. (1987) Plant Physiol. 84:965-968; Romer et al. (1993) Biochem. Biophys. Res. Commun. 196: 1414-1421 ; and Shah et al. (1986) Science 233:478-481. For example, a chloroplast transit peptide known in the art can be fused to the amino acid sequence of a TK

polypeptide of the invention by operably linking a choloroplast-targeting sequence to the 5'- end of a nucleotide sequence encoding the TK polypeptide.

Chloroplast targeting sequences are known in the art and include the chloroplast small subunit of ribulose-l,5-bisphosphate carboxylase (Rubisco) (de Castro Silva Filho et al. (1996) Plant Mol. Biol. 30:769-780; Schnell et al. (1991 ) J Biol. Chem. 266(5):3335-3342); EPSPS (Archer et al. (1990) J Bioenerg. Biomemb. 22(6):789-810); tryptophan synthase (Zhao et al. (1995) J Biol. Chem. 270(1 1 ):6081 -6087); plastocyanin (Lawrence et al. (1997) J Biol. Chem. 272(33):20357-20363); chorismate synthase (Schmidt et al. (1993) J Biol. Chem. 268(36):27447-27457); and the light harvesting chlorophyll a/b binding protein

(LHBP) (Lamppa et al. (1988) J Biol. Chem. 263: 14996-14999). See also Von Heijne et al. (1991 ) Plant Mol. Biol. Rep. 9: 104- 126; Clark et al. (1989) J Biol. Chem. 264: 17544- 17550; Della-Cioppa et al. (1987) Plant Physiol. 84:965-968; Romer et al. (1993) Biochem Biophys. Res. Commun. 196: 1414-1421 ; and Shah et al. (1986) Science 233:478-481.

Methods for transformation of chloroplasts are known in the art. See, for example, Svab et al. (1990) Proc. Natl. Acad. ScL USA 87:8526-8530; Svab and Maliga (1993) Proc. Natl. Acad. Sci. USA 90:913-917; Svab and Maliga (1993) EMBO J. 12:601 -606. The method relies on particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination. Additionally, plastid transformation can be accomplished by transactivation of a silent plastid-borne transgene by tissue-preferred expression of a nuclear-encoded and plastid-directed RNA polymerase. Such a system has been reported in McBride et al. (1994) Proc. Natl. Acad. Sci. USA 91 :7301 - 7305.

The nucleic acids of interest to be targeted to the chloroplast may be optimized for expression in the chloroplast to account for differences in codon usage between the plant nucleus and this organelle. In this manner, the nucleic acids of interest may be synthesized using chloroplast-preferred codons. See, for example, U.S. Patent No. 5,380,831 , herein incorporated by reference. Numerous plant transformation vectors and methods for transforming plants are available.

See, for example, An, G. et al. (1986) Plant PysioL, 81 :301 -305; Fry, J., et al. ( 1987) Plant

Cell Rep. 6:321 -325; Block, M. (1988) Theor. Appl. Genet .16: 161 -1 1 A; Hinchee, et al.

(1990) Stadler. Genet. Symp.2032\2.203-2\2; Cousins, et al. (1991 ) Aust. J. Plant Physiol.

18:481-494; Chee, P. P. and Slightom, J. L. (1992) Gene.l I 8:255-260; Christou, et al. (1992) Trends. Biotechnol. 10:239-246; Halluin, et al. (1992) Bio/Technol. 10:309-314; Dhir, et al. (1992) Plant Physiol. 99:81 -88; Casas et al. (1993) Proc. Nat. Acad Sd. USA 90: 1

1212-1 1216; Christou, P. (1993) In Vitro Cell. Dev. Biol.-Plant; 29P.119-124; Davies, et al.

(1993) Plant Cell Rep. 12: 180-183; Dong, J. A. and Mchughen, A. (1993) Plant ScL 91 :

139-148; Franklin, C. I. and Trieu, T. N. (1993) Plant. Physiol. 102: 167; Golovkin, et al. (1993) Plant ScL 90:41 -52; Guo Chin ScL Bull. 38:2072-2078; Asano, et al. (1994) Plant

Cell Rep. 13; Ayeres N. M. and Park, W. D. (1994) Crit. Rev. Plant. Sci. 13:219-239;

Barcelo, et al. (1994) Plant. J. 5:583-592; Becker, et al. (1994) Plant. J. 5:299-307;

Borkowska et al. (1994) Acta. Physiol Plant. 16:225-230; Christou, P. (1994) Agro. Food.

Ind. Hi Tech. 5: 17-27; Eapen et al. (1994) Plant Cell Rep. 13:582-586; Hartman, et al. (1994) Bio-Technology 12: 919923; Ritala, et al. (1994) Plant. Mol. Biol. 24:317-325; and

Wan, Y. C. and Lemaux, P. G. (1994) Plant Physiol. 104:3748.

In some embodiments, the methods of the invention involve introducing a polynucleotide construct into a plant. By "introducing" is intended presenting to the plant the polynucleotide construct in such a manner that the construct gains access to the interior of a cell of the plant. The methods of the invention do not depend on a particular method for introducing a polynucleotide construct to a plant, only that the polynucleotide construct gains access to the interior of at least one cell of the plant. Methods for introducing polynucleotide constructs into plants are known in the art including, but not limited to, stable transformation methods, transient transformation methods, and virus-mediated methods. The term

"introduction" or "transformation" as referred to herein further means the transfer of an exogenous polynucleotide into a host cell, irrespective of the method used for transfer. Plant tissue capable of subsequent clonal propagation, whether by organogenesis or embryogenesis, may be transformed with a genetic construct of the present invention and a whole plant regenerated there from. The particular tissue chosen will vary depending on the clonal propagation systems available for, and best suited to, the particular species being transformed. Exemplary tissue targets include leaf disks, pollen, embryos, cotyledons, hypocotyls, megagametophytes, callus tissue, existing meristematic tissue (e.g., apical meristem, axillary buds, and root meristems), and induced meristem tissue (e.g., cotyledon meristem and hypocotyl meristem). The polynucleotide may be transiently or stably introduced into a host cell and may be maintained non-integrated, for example, as a plasmid. Alternatively, it may be integrated into the host genome. The resulting transformed plant cell may then be used to regenerate a transformed plant in a manner known to persons skilled in the art.

By "stable transformation" is intended that the polynucleotide construct introduced into a plant integrates into the genome of the plant and is capable of being inherited by

descendent thereof. By "transient transformation" is intended that a polynucleotide construct introduced into a plant does not integrate into the genome of the plant.

For the transformation of plants and plant cells, the nucleotide sequences of the invention are inserted using standard techniques into any vector known in the art that is suitable for expression of the nucleotide sequences in a plant or plant cell. The selection of the vector depends on the preferred transformation technique and the target plant species to be transformed. In an embodiment of the invention, the encoding nucleotide sequence is operably linked to a plant promoter, e.g. a promoter known in the art for high-level expression in a plant cell, and this construct is then introduced into a plant cell that is susceptible to TK-inhibiting herbicides; and a transformed plant is regenerated. In some embodiments, the transformed plant is tolerant to exposure to a level of TK-inhibiting herbicides that would kill or significantly injure a plant regenerated from an untransformed cell. This method can be applied to any plant species or crops. Methodologies for constructing plant expression vectors and introducing foreign nucleic acids into plants are generally known in the art. For example, foreign DNA can be introduced into plants, using tumor-inducing (Ti) plasmid vectors. Other methods utilized for foreign DNA delivery involve the use of PEG mediated protoplast transformation, electroporation, microinjection whiskers, and biolistics or microprojectile bombardment for direct DNA uptake. Such methods are known in the art. (U.S. Pat. No. 5,405,765 to Vasil et al.; Bilang et a (1991 ) Gene 100: 247-250; Scheid et al.al, (1991) MoL Gen. Genet., 228: 104- 1 12; Guerche et al., (1987) Plant Science 52: 1 1 1 -1 16; Neuhause et al., (1987) Theor. Appl Genet. 75: 30-36; Klein et al., (1987) Nature 327: 70-73; Howell et al., (1980) Science 208: 1265; Horsch et al., (1985) Science 227: 1229-1231 ; DeBlock et al., (1989) Plant Physiology 91 : 694-701 ; Methods for Plant Molecular Biology (Weissbach and Weissbach, eds.) Academic Press, Inc. (1988) and Methods in Plant Molecular Biology (Schuler and Zielinski, eds.) Academic Press, Inc. (1989).

Other suitable methods of introducing nucleotide sequences into plant cells include microinjection as described by, e.g., Crossway et al. (1986) Biotechniques 4:320-334, electroporation as described by e.g., Riggs et al. (1986) Proc. Natl. Acad. ScL USA

83:5602- 5606, Agrobacterium-mediated transformation as described by e.g., Townsend et al., U.S. Patent No. 5,563,055, Zhao et al., U.S. Patent No. 5,981 ,840, direct gene transfer as described by, e.g., Paszkowski et al. (1984) EMBO J. 3:2717-2722, and ballistic particle acceleration as described by, e.g., U.S. Patent Nos. 4,945,050; 5,879,918; 5,886,244; and 5,932,782; Tomes et al. (1995) "Direct DNA Transfer into Intact Plant Cells via

Microprojectile Bombardment," in Plant Cell, Tissue, and Organ Culture: Fundamental Methods, ed. Gamborg and Phillips (Springer- Verlag, Berlin); McCabe et al. (1988) Biotechnology 6:923-926); and Led transformation (WO 00/28058). Also see, Weissinger et al., (1988) Ann. Rev. Genet. 22:421 -477; Sanford et al, (1987) Particulate Science and Technology 5:27-37 (onion); Christou et al, (1988) Plant Physiol. 87:671 -674 (soybean); McCabe et al., (1988) Bio/Technology 6:923-926 (soybean); Finer and McMullen (1991 ) In Vitro Cell Dev. Biol. 27P: 175-182 (soybean); Singh et al, (1998) Theor. Appl. Genet.

96:319-324 (soybean); Datta et al., (1990) Biotechnology 8:736-740 (rice); Klein et al., (1988) PNAS, 85:4305-4309 (maize); Klein et al., (1988) Biotechnology 6:559-563 (maize); U.S. Patent Nos. 5,240,855; 5,322,783; and 5,324,646; Tomes et al., (1995) "Direct DNA Transfer into Intact Plant Cells via Microprojectile Bombardment," in Plant Cell, Tissue, and Organ Culture: Fundamental Methods, ed. Gamborg (Springer- Verlag, Berlin) (maize); Klein et al., (1988) Plant Physiol. 91 :440-444 (maize); Fromm et al., (1990) Biotechnology 8:833-839 (maize); Hooykaas-Van Slogteren et al., (1984) Nature (London) 31 1 :763-764; Bowen et al, U.S. Patent No. 5,736,369 (cereals); Bytebier et al, (1987) PNAS 84:5345- 5349 (Liliaceae); De Wet et al., (1985) in The Experimental Manipulation of Ovule Tissues, ed. Chapman et al, (Longman, New York), pp. 197-209 (pollen); Kaeppler et al., (1990) Plant Cell Reports 9:415-418 and Kaeppler et al., (1992) Theor. Apph Genet. 84:560-566 (whisker-mediated transformation); D'Halluin et al., (1992) Plant Cell 4: 1495-1505

(electroporation); Li et al., (1993) Plant Cell Reports 12:250- 255 and Christou and Ford (1995) Annals of Botany 75:407-413 (rice); Osjoda et al, (1996) Nature Biotechnology

14:745-750 (maize via Agrobacterium tumefaciens); each of which is herein incorporated by reference.

Transgenic plants, including transgenic crop plants, are preferably produced via Agrobacterium- mediated transformation. An advantageous transformation method is the transformation in planta. To this end, it is possible, for example, to allow the agrobacteria to act on plant seeds or to inoculate the plant meristem with agrobacteria. It has proved particularly expedient in accordance with the invention to allow a suspension of transformed agrobacteria to act on the intact plant or at least on the flower primordia. The plant is subsequently grown on until the seeds of the treated plant are obtained (Clough and Bent, Plant J. (1998) 16, 735-743).

Methods for Agrobacterium-mediated transformation of rice include well known methods for rice transformation, such as those described in any of the following: European patent application EP 1 198985 A1 , Aldemita and Hodges (Planta 199: 612-617, 1996); Chan et al. (Plant Mol Biol 22 (3): 491 -506, 1993), Hiei et al. (Plant J 6 (2): 271 -282, 1994), which disclosures are

incorporated by reference herein as if fully set forth. In the case of corn transformation, the preferred method is as described in either Ishida et al. (Nat. Biotechnol 14(6): 745-50, 1996) or Frame et al. (Plant Physiol 129(1 ): 13-22, 2002), which disclosures are incorporated by reference herein as if fully set forth. Said methods are further described by way of example in B. Jenes et al., Techniques for Gene Transfer, in: Transgenic Plants, Vol. 1 , Engineering and Utilization, eds. S.D. Kung and R. Wu, Academic Press (1993) 128-143 and in Potrykus Annu. Rev. Plant Physiol. Plant Molec. Biol. 42 (1991 ) 205-225). The nucleic acids or the construct to be expressed is preferably cloned into a vector, which is suitable for transforming Agrobacterium tumefaciens, for example pBin19 (Bevan et al., Nucl. Acids Res. 12 (1984) 871 1 ). Agrobacteria transformed by such a vector can then be used in known manner for the transformation of plants, such as plants used as a model, like Arabidopsis (Arabidopsis thaliana is within the scope of the present invention not considered as a crop plant), or crop plants such as, by way of example, tobacco plants, for example by immersing bruised leaves or chopped leaves in an agrobacterial solution and then culturing them in suitable media. The

transformation of plants by means of Agrobacterium tumefaciens is described, for example, by Hofgen and Willmitzer in Nucl. Acid Res. (1988) 16, 9877 or is known inter alia from F.F. White, Vectors for Gene Transfer in Higher Plants; in Transgenic Plants, Vol. 1 , Engineering and Utilization, eds. S.D. Kung and R. Wu, Academic Press, 1993, pp. 15-38.

One transformation method known to those of skill in the art is the dipping of a flowering plant into an Agrobacteria solution, wherein the Agrobacteria contains the TK nucleic acid, followed by breeding of the transformed gametes. Agrobacterium mediated plant transformation can be performed using for example the GV3101 (pMP90) (Koncz and Schell, 1986, Mol. Gen. Genet. 204:383-396) or LBA4404 (Clontech) Agrobacterium tumefaciens strain. Transformation can be performed by standard transformation and regeneration techniques (Deblaere et al., 1994, Nucl. Acids. Res. 13:4777-4788; Gelvin, Stanton B. and Schilperoort, Robert A, Plant Molecular Biology Manual, 2nd Ed. - Dordrecht : Kluwer Academic Publ., 1995. - in Sect., Ringbuc

Zentrale Signatur: BT1 1 -P ISBN 0-7923-2731 -4; Glick, Bernard R. and Thompson, John E., Methods in Plant Molecular Biology and Biotechnology, Boca Raton : CRC Press, 1993 360 S., ISBN 0-8493-5164-2). For example, rapeseed can be transformed via cotyledon or hypocotyl transformation (Moloney et al., 1989, Plant Cell Report 8:238-242; De Block et al., 1989, Plant Physiol. 91 :694-701 ). Use of antibiotics for Agrobacterium and plant selection depends on the binary vector and the Agrobacterium strain used for transformation. Rapeseed selection is normally performed using kanamycin as selectable plant marker. Agrobacterium mediated gene transfer to flax can be performed using, for example, a technique described by Mlynarova et al., 1994, Plant Cell Report 13:282-285. Additionally, transformation of soybean can be performed using for example a technique described in European Patent No. 0424 047, U.S. Patent No. 5,322,783, European Patent No. 0397 687, U.S. Patent No. 5,376,543, or U.S. Patent No.

5,169,770. Transformation of maize can be achieved by particle bombardment, polyethylene glycol mediated DNA uptake, or via the silicon carbide fiber technique. (See, for example, Freeling and Walbot "The maize handbook" Springer Verlag: New York (1993) ISBN 3-540- 97826-7). A specific example of maize transformation is found in U.S. Patent No. 5,990,387, and a specific example of wheat transformation can be found in PCT Application No. WO 93/07256.

In some embodiments, polynucleotides of the present invention may be introduced into plants by contacting plants with a virus or viral nucleic acids. Generally, such methods involve incorporating a polynucleotide construct of the invention within a viral DNA or RNA molecule. It is recognized that the polypeptides of the invention may be initially synthesized as part of a viral polyprotein, which later may be processed by proteolysis in vivo or in vitro to produce the desired recombinant polypeptide. Further, it is recognized that promoters of the invention also encompass promoters utilized for transcription by viral RNA polymerases. Methods for introducing polynucleotide constructs into plants and expressing a protein encoded therein, involving viral DNA or RNA molecules, are known in the art. See, for example, U.S. Patent Nos. 5,889,191 , 5,889,190, 5,866,785, 5,589,367 and 5,316,931 ; herein incorporated by reference. The cells that have been transformed may be grown into plants in accordance with conventional ways. See, for example, McCormick et a (1986) Plant Cell Reports 5:81 -84. These plants may then be grown, and either pollinated with the same transformed strain or different strains, and the resulting hybrid having constitutive expression of the desired phenotypic characteristic identified. Two or more generations may be grown to ensure that expression of the desired phenotypic characteristic is stably maintained and inherited and then seeds harvested to ensure expression of the desired phenotypic characteristic has been achieved.

The present invention may be used for transformation of any plant species, including, but not limited to, monocots and dicots. Examples of plant species of interest include, but are not limited to, corn or maize (Zea mays), Brassica sp. (e.g., B. napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceum), foxtail millet (Setaria italica), finger millet (Eleusine coracana)), sunflower (Helianthus annu ), saffiower (Carthamus tinctorius), wheat (Triticum aestivum, T. Turgidum ssp. durum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solarium tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassava (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifera), pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa

(Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley, vegetables, ornamentals, and conifers.

Preferably, plants of the present invention are crop plants (for example, sunflower, Brassica sp., cotton, sugar, beet, soybean, peanut, alfalfa, saffiower, tobacco, corn, rice, wheat, rye, barley triticale, sorghum, millet, etc.). In addition to the transformation of somatic cells, which then have to be regenerated into intact plants, it is also possible to transform the cells of plant meristems and in particular those cells which develop into gametes. In this case, the transformed gametes follow the natural plant development, giving rise to transgenic plants. Thus, for example, seeds of Arabidopsis are treated with agrobacteria and seeds are obtained from the developing plants of which a certain proportion is transformed and thus transgenic [Feldman, KA and Marks MD (1987). Mol Gen Genet 208:274-289; Feldmann K (1992). In: C Koncz, N-H Chua and J Shell, eds, Methods in Arabidopsis Research. Word Scientific, Singapore, pp. 274-289]. Alternative methods are based on the repeated removal of the inflorescences and incubation of the excision site in the center of the rosette with transformed agrobacteria, whereby transformed seeds can likewise be obtained at a later point in time (Chang (1994). Plant J. 5: 551 -558; Katavic (1994). Mol Gen Genet, 245: 363-370). However, an especially effective method is the vacuum infiltration method with its modifications such as the "floral dip" method. In the case of vacuum infiltration of Arabidopsis, intact plants under reduced pressure are treated with an agrobacterial suspension [Bechthold, N (1993). C R Acad Sci Paris Life Sci, 316: 1 194-1 199], while in the case of the "floral dip" method the developing floral tissue is incubated briefly with a surfactant-treated agrobacterial suspension [Clough, SJ and Bent AF (1998) The Plant J. 16, 735-743]. A certain proportion of transgenic seeds are harvested in both cases, and these seeds can be distinguished from non-transgenic seeds by growing under the above-described selective conditions. In addition the stable transformation of plastids is of advantages because plastids are inherited maternally is most crops reducing or eliminating the risk of transgene flow through pollen. The transformation of the chloroplast genome is generally achieved by a process which has been schematically displayed in Klaus et al., 2004 [Nature Biotechnology 22 (2), 225-229]. Briefly the

sequences to be transformed are cloned together with a selectable marker gene between flanking sequences homologous to the chloroplast genome. These homologous flanking sequences direct site specific integration into the plastome. Plastidal transformation has been described for many different plant species and an overview is given in Bock (2001 )

Transgenic plastids in basic research and plant biotechnology. J Mol Biol. 2001 Sep 21 ; 312 (3):425-38 or Maliga, P (2003) Progress towards commercialization of plastid

transformation technology. Trends Biotechnol. 21 , 20-28. Further biotechnological progress has recently been reported in form of marker free plastid transformants, which can be produced by a transient co-integrated maker gene (Klaus et al., 2004, Nature Biotechnology 22(2), 225-229). The genetically modified plant cells can be regenerated via all methods with which the skilled worker is familiar. Suitable methods can be found in the

abovementioned publications by S.D. Kung and R. Wu, Potrykus or Hofgen and Willmitzer. Generally after transformation, plant cells or cell groupings are selected for the presence of one or more markers which are encoded by plant-expressible genes co-transferred with the gene of interest, following which the transformed material is regenerated into a whole plant. To select transformed plants, the plant material obtained in the transformation is, as a rule, subjected to selective conditions so that transformed plants can be distinguished from untransformed plants. For example, the seeds obtained in the above-described manner can be planted and, after an initial growing period, subjected to a suitable selection by spraying. A further possibility consists in growing the seeds, if appropriate after sterilization, on agar plates using a suitable selection agent so that only the transformed seeds can grow into plants. Alternatively, the transformed plants are screened for the presence of a selectable marker such as the ones described above.

Following DNA transfer and regeneration, putatively transformed plants may also be evaluated, for instance using Southern analysis, for the presence of the gene of interest, copy number and/or genomic organisation. Alternatively or additionally, expression levels of the newly introduced DNA may be monitored using Northern and/or Western analysis, both techniques being well known to persons having ordinary skill in the art.

The generated transformed plants may be propagated by a variety of means, such as by clonal propagation or classical breeding techniques. For example, a first generation (or T1 ) transformed plant may be selfed and homozygous second-generation (or T2) transformants selected, and the T2 plants may then further be propagated through classical breeding techniques. The generated transformed organisms may take a variety of forms. For example, they may be chimeras of transformed cells and non -transformed cells; clonal transformants (e.g., all cells transformed to contain the expression cassette); grafts of transformed and untransformed tissues (e.g., in plants, a transformed rootstock grafted to an untransformed scion).

Preferably, the expression of the nucleic acid in the plant results in the plant's increased resistance to TK-inhibiting herbicide as compared to a wild type variety of the plant.

In another embodiment, the invention refers to a plant, comprising a plant cell according to the present invention, wherein expression of the nucleic acid in the plant results in the plant's increased resistance to TK-inhibiting herbicide as compared to a wild type variety of the plant.

The plants described herein can be either transgenic crop plants or non-transgenic plants. In addition to the general definition, give SUPRA, "transgenic", "transgene" or "recombinant" means with regard to, for example, a nucleic acid sequence, an expression cassette, gene construct or a vector comprising the nucleic acid sequence or an organism transformed with the nucleic acid sequences, expression cassettes or vectors according to the invention, all those constructions brought about by recombinant methods in which either

(a) the nucleic acid sequences encoding proteins useful in the methods of the invention, or

(b) genetic control sequence(s) which is operably linked with the nucleic acid sequence

according to the invention, for example a promoter, or

(c) a) and b)

are not located in their natural genetic environment or have been modified by recombinant methods, it being possible for the modification to take the form of, for example, a substitution, addition, deletion, inversion or insertion of one or more nucleotide residues in order to allow for the expression of the mutated TK of the present invention. The natural genetic environment is understood as meaning the natural genomic or chromosomal locus in the original plant or the presence in a genomic library. In the case of a genomic library, the natural genetic environment of the nucleic acid sequence is preferably retained, at least in part. The environment flanks the nucleic acid sequence at least on one side and has a sequence length of at least 50 bp, preferably at least 500 bp, especially preferably at least 1000 bp, most preferably at least 5000 bp. A naturally occurring expression cassette - for example the naturally occurring combination of the natural promoter of the nucleic acid sequences with the corresponding nucleic acid sequence encoding a polypeptide useful in the methods of the present invention, as defined above - becomes a transgenic expression cassette when this expression cassette is modified by non-natural, synthetic ("artificial") methods such as, for example, mutagenic treatment.

Suitable methods are described, for example, in US 5,565,350 or WO 00/15815.

A transgenic plant for the purposes of the invention is thus understood as meaning, as above, that the nucleic acids of the invention are not at their natural locus in the genome of said plant, it being possible for the nucleic acids to be expressed homologously or heterologously. However, as mentioned, transgenic also means that, while the nucleic acids according to the invention or used in the inventive method are at their natural position in the genome of a plant, the sequence has been modified with regard to the natural sequence, and/or that the regulatory sequences of the natural sequences have been modified. Transgenic is preferably understood as meaning the expression of the nucleic acids according to the invention at an unnatural locus in the genome, i.e. homologous or, preferably, heterologous expression of the nucleic acids takes place.

Preferred transgenic plants are mentioned herein. Furthermore, the term "transgenic" refers to any plant, plant cell, callus, plant tissue, or plant part, that contains all or part of at least one recombinant polynucleotide. In many cases, all or part of the recombinant polynucleotide is stably integrated into a chromosome or stable extra-chromosomal element, so that it is passed on to successive generations. For the purposes of the invention, the term "recombinant polynucleotide" refers to a polynucleotide that has been altered, rearranged, or modified by genetic engineering. Examples include any cloned polynucleotide, or polynucleotides, that are linked or joined to heterologous sequences. The term "recombinant" does not refer to alterations of polynucleotides that result from naturally occurring events, such as spontaneous mutations, or from non-spontaneous mutagenesis followed by selective breeding.

Plants containing mutations arising due to non-spontaneous mutagenesis and selective breeding are referred to herein as non-transgenic plants and are included in the present invention. In embodiments wherein the plant is transgenic and comprises multiple mutated TK nucleic acids, the nucleic acids can be derived from different genomes or from the same genome. Alternatively, in embodiments wherein the plant is non-transgenic and comprises multiple mutated TK nucleic acids, the nucleic acids are located on different genomes or on the same genome. In certain embodiments, the present invention involves herbidicide-resistant plants that are produced by mutation breeding. Such plants comprise a polynucleotide encoding a mutated TK and are tolerant to one or more TK-inhibiting herbicides. Such methods can involve, for example, exposing the plants or seeds to a mutagen, particularly a chemical mutagen such as, for example, ethyl methanesulfonate (EMS) and selecting for plants that have enhanced tolerance to at least one or more TK-inhibiting herbicide.

However, the present invention is not limited to herbicide-tolerant plants that are produced by a mutagenesis method involving the chemical mutagen EMS. Any mutagenesis method known in the art may be used to produce the herbicide-resistant plants of the present invention. Such mutagenesis methods can involve, for example, the use of any one or more of the following mutagens: radiation, such as X-rays, Gamma rays (e.g., cobalt 60 or cesium 137), neutrons, (e.g., product of nuclear fission by uranium 235 in an atomic reactor), Beta radiation (e.g., emitted from radioisotopes such as phosphorus 32 or carbon 14), and ultraviolet radiation (preferably from 2500 to 2900 nm), and chemical mutagens such as base analogues (e.g., 5- bromo-uracil), related compounds (e.g., 8-ethoxy caffeine), antibiotics (e.g., streptonigrin), alkylating agents (e.g., sulfur mustards, nitrogen mustards, epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, or acridines. Herbicide- resistant plants can also be produced by using tissue culture methods to select for plant cells comprising herbicide-resistance mutations and then regenerating herbicide-resistant plants therefrom. See, for example, U.S. Patent Nos. 5,773,702 and 5,859,348, both of which are herein incorporated in their entirety by reference. Further details of mutation breeding can be found in "Principals of Cultivar Development" Fehr, 1993 Macmillan Publishing Company the disclosure of which is incorporated herein by reference

The plant of the present invention comprises at least one mutated TK nucleic acid or over- expressed wild-type TK nucleic acid, and has increased tolerance to a TK-inhibiting herbicide as compared to a wild-type variety of the plant. It is possible for the plants of the present invention to have multiple wild-type or mutated TK nucleic acids from different genomes since these plants can contain more than one genome. For example, a plant contains two genomes, usually referred to as the A and B genomes. Because TK is a required metabolic enzyme, it is assumed that each genome has at least one gene coding for the TK enzyme (i.e. at least one TK gene). As used herein, the term "TK gene locus" refers to the position of an TK gene on a genome, and the terms "TK gene" and "TK nucleic acid" refer to a nucleic acid encoding the TK enzyme. The TK nucleic acid on each genome differs in its nucleotide sequence from an TK nucleic acid on another genome. One of skill in the art can determine the genome of origin of each TK nucleic acid through genetic crossing and/or either sequencing methods or exonuclease digestion methods known to those of skill in the art.

The present invention includes plants comprising one, two, three, or more mutated TK alleles, wherein the plant has increased tolerance to a TK-inhibiting herbicide as compared to a wild- type variety of the plant. The mutated TK alleles can comprise a nucleotide sequence selected from the group consisting of a polynucleotide as defined in SEQ ID NO: 182 or 183, or a variant or derivative thereof, a polynucleotide encoding a polypeptide as defined in SEQ ID NO: 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 1 10, 1 1 1 , 1 12, 1 13, 1 14, 1 15, 1 16, 1 17, 1 18, 1 19, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181 , or a variant or derivative, homologue, orthologue, paralogue thereof, a polynucleotide comprising at least 60 consecutive nucleotides of any of the aforementioned polynucleotides; and a polynucleotide complementary to any of the aforementioned polynucleotides.

"Alleles" or "allelic variants" are alternative forms of a given gene, located at the same chromosomal position. Allelic variants encompass Single Nucleotide Polymorphisms (SNPs), as well as Small Insertion/Deletion Polymorphisms (INDELs). The size of INDELs is usually less than 100 bp. SNPs and INDELs form the largest set of sequence variants in naturally occurring polymorphic strains of most organisms

The term "variety" refers to a group of plants within a species defined by the sharing of a common set of characteristics or traits accepted by those skilled in the art as sufficient to distinguish one cultivar or variety from another cultivar or variety. There is no implication in either term that all plants of any given cultivar or variety will be genetically identical at either the whole gene or molecular level or that any given plant will be homozygous at all loci. A cultivar or variety is considered "true breeding" for a particular trait if, when the true-breeding cultivar or variety is self-pollinated, all of the progeny contain the trait. The terms "breeding line" or "line" refer to a group of plants within a cultivar defined by the sharing of a common set of

characteristics or traits accepted by those skilled in the art as sufficient to distinguish one breeding line or line from another breeding line or line. There is no implication in either term that all plants of any given breeding line or line will be genetically identical at either the whole gene or molecular level or that any given plant will be homozygous at all loci. A breeding line or line is considered "true breeding" for a particular trait if, when the true-breeding line or breeding line is self-pollinated, all of the progeny contain the trait. In the present invention, the trait arises from a mutation in a TK gene of the plant or seed.

The herbicide-resistant plants of the invention that comprise polynucleotides encoding mutated TK polypeptides also find use in methods for increasing the herbicide-resistance of a plant through conventional plant breeding involving sexual reproduction. The methods comprise crossing a first plant that is a herbicide-resistant plant of the invention to a second plant that may or may not be resistant to the same herbicide or herbicides as the first plant or may be resistant to different herbicide or herbicides than the first plant. The second plant can be any plant that is capable of producing viable progeny plants (i.e., seeds) when crossed with the first plant. Typically, but not necessarily, the first and second plants are of the same species. The methods can optionally involve selecting for progeny plants that comprise the mutated TK polypeptides of the first plant and the herbicide resistance characteristics of the second plant. The progeny plants produced by this method of the present invention have increased resistance to a herbicide when compared to either the first or second plant or both. When the first and second plants are resistant to different herbicides, the progeny plants will have the combined herbicide tolerance characteristics of the first and second plants. The methods of the invention can further involve one or more generations of backcrossing the progeny plants of the first cross to a plant of the same line or genotype as either the first or second plant. Alternatively, the progeny of the first cross or any subsequent cross can be crossed to a third plant that is of a different line or genotype than either the first or second plant. The present invention also provides plants, plant organs, plant tissues, plant cells, seeds, and non-human host cells that are transformed with the at least one polynucleotide molecule, expression cassette, or transformation vector of the invention. Such transformed plants, plant organs, plant tissues, plant cells, seeds, and non-human host cells have enhanced tolerance or resistance to at least one herbicide, at levels of the herbicide that kill or inhibit the growth of an untransformed plant, plant tissue, plant cell, or non-human host cell, respectively. Preferably, the transformed plants, plant tissues, plant cells, and seeds of the invention are Arabidopsis thaliana and crop plants.

It is to be understood that the plant of the present invention can comprise a wild type TK nucleic acid in addition to a mutated TK nucleic acid. It is contemplated that the TK-inhibiting herbicide tolerant lines may contain a mutation in only one of multiple TK isoenzymes. Therefore, the present invention includes a plant comprising one or more mutated TK nucleic acids in addition to one or more wild type TK nucleic acids.

In another embodiment, the invention refers to a seed produced by a transgenic plant comprising a plant cell of the present invention, wherein the seed is true breeding for an increased resistance to a TK-inhibiting herbicide as compared to a wild type variety of the seed.

In other aspects, TK-inhibiting herbicides-tolerant plants of the present invention can be employed as TK-inhibiting herbicides-tolerance trait donor lines for development, as by traditional plant breeding, to produce other varietal and/or hybrid crops containing such trait or traits. All such resulting variety or hybrids crops, containing the ancestral TK-inhibiting herbicides-tolerance trait or traits can be referred to herein as progeny or descendant of the ancestral, TK-inhibiting herbicides-tolerant line(s).

In other embodiments, the present invention provides a method for producing a TK- inhibiting herbicides-tolerant plant. The method comprises: crossing a first TK-inhibiting herbicides-tolerant plant with a second plant to produce a TK-inhibiting herbicides-tolerant progeny plant, wherein the first plant and the progeny plant comprise in at least some of their cells a polynucleotide operably linked to a promoter operable in plant cells, the recombinant polynucleotide being effective in the cells of the first plant to express a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

In some embodiments, traditional plant breeding is employed whereby the TK-inhibiting herbicides-tolerant trait is introduced in the progeny plant resulting therefrom. In one embodiment, the present invention provides a method for producing a TK-inhibiting herbicides-tolerant progeny plant, the method comprising: crossing a parent plant with a TK-inhibiting herbicides-tolerant plant to introduce the TK-inhibiting herbicides-tolerance characteristics of the TK-inhibiting herbicides-tolerant plant into the germplasm of the progeny plant, wherein the progeny plant has increased tolerance to the TK-inhibiting herbicides relative to the parent plant. In other embodiments, the method further comprises the step of introgressing the TK-inhibiting herbicides-tolerance characteristics through traditional plant breeding techniques to obtain a descendent plant having the TK-inhibiting herbicides-tolerance characteristics.

In other aspects, plants of the invention include those plants which, in addition to being TK- inhibiting herbicides-tolerant, have been subjected to further genetic modifications by breeding, mutagenesis or genetic engineering, e.g. have been rendered tolerant to applications of specific other classes of herbicides, such as AHAS inhibitors; auxinic herbicides; bleaching herbicides such as hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors or phytoene desaturase (PDS) inhibitors; EPSPS inhibitors such as glyphosate; glutamine synthetase (GS) inhibitors such as glufosinate; lipid biosynthesis inhibitors such as acetyl CoA carboxylase (ACCase) inhibitors; or oxynil {i.e. bromoxynil or ioxynil) herbicides as a result of conventional methods of breeding or genetic engineering, Thus, TK-inhibiting herbicides-tolerant plants of the invention can be made resistant to multiple classes of herbicides through multiple genetic modifications, such as resistance to both glyphosate and glufosinate or to both glyphosate and a herbicide from another class such as HPPD inhibitors, AHAS inhibitors, or ACCase inhibitors. These herbicide resistance technologies are, for example, described in Pest Management Science (at volume, year, page): 61 , 2005, 246; 61 , 2005, 258; 61 , 2005, 277; 61 , 2005, 269; 61 , 2005, 286; 64, 2008, 326; 64, 2008, 332; Weed Science 57, 2009, 108; Australian Journal of Agricultural

Research 58, 2007, 708; Science 316, 2007, 1 185; and references quoted therein. For example, TK-inhibiting herbicides-tolerant plants of the invention, in some embodiments, may be tolerant to ACCase inhibitors, such as "dims" {e.g., cycloxydim, sethoxydim, clethodim, or tepraloxydim), "fops" {e.g. , clodinafop, diclofop, fluazifop, haloxyfop, or quizalofop), and "dens" (such as pinoxaden); to auxinic herbicides, such as dicamba; to EPSPS inhibitors, such as glyphosate; to other TK inhibitors; and to GS inhibitors, such as glufosinate.

In addition to these classes of inhibitors, TK-inhibiting herbicides-tolerant plants of the invention may also be tolerant to herbicides having other modes of action, for example, chlorophyll/carotenoid pigment inhibitors, cell membrane disrupters, photosynthesis inhibitors, cell division inhibitors, root inhibitors, shoot inhibitors, and combinations thereof.

Such tolerance traits may be expressed, e.g. : as mutant or wildtype HPPD proteins, as mutant AHASL proteins, mutant ACCase proteins, mutant EPSPS proteins, or mutant glutamine synthetase proteins; or as mutant native, inbred, or transgenic aryloxyalkanoate dioxygenase (AAD or DHT), haloarylnitrilase (BXN), 2,2-dichloropropionic acid

dehalogenase (DEH), glyphosate-N- acetyltransferase (GAT), glyphosate decarboxylase (GDC), glyphosate oxidoreductase (GOX), glutathione-S-transferase (GST),

phosphinothricin acetyltransferase (PAT or bar), or CYP450s proteins having an herbicide- degrading activity. TK-inhibiting herbicides- tolerant plants hereof can also be stacked with other traits including, but not limited to, pesticidal traits such as Bt Cry and other proteins having pesticidal activity toward coleopteran, lepidopteran, nematode, or other pests;

nutrition or nutraceutical traits such as modified oil content or oil profile traits, high protein or high amino acid concentration traits, and other trait types known in the art.

Furthermore, in other embodiments, TK-inhibiting herbicides-tolerant plants are also covered which are, by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such characteristics, rendered able to synthesize one or more insecticidal proteins, especially those known from the bacterial genus Bacillus, particularly from Bacillus thuringiensis, such as [delta]-endotoxins, e.g. CrylA(b), CrylA(c), CrylF, CrylF(a2), CryllA(b), CrylllA, CrylllB(bl) or Cry9c; vegetative insecticidal proteins (VIP), e.g. VIP1 , VIP2, VIP3 or VIP3A; insecticidal proteins of bacteria colonizing nematodes, e.g. Photorhabdus spp. or Xenorhabdus spp.; toxins produced by animals, such as scorpion toxins, arachnid toxins, wasp toxins, or other insect-specific neurotoxins; toxins produced by fungi, such streptomycete toxins; plant lectins, such as pea or barley lectins; agglutinins; proteinase inhibitors, such as trypsin inhibitors, serine protease inhibitors, patatin, cystatin or papain inhibitors; ribosome-inactivating proteins (RIP), such as ricin, maize-RIP, abrin, luffin, saporin or bryodin; steroid metabolism enzymes, such as 3-hydroxy-steroid oxidase, ecdysteroid-IDP-glycosyl-transferase, cholesterol oxidases, ecdysone inhibitors or HMG- CoA-reductase; ion channel blockers, such as blockers of sodium or calcium channels; juvenile hormone esterase; diuretic hormone receptors (helicokinin receptors); stilben synthase, bibenzyl synthase, chitinases or glucanases. In the context of the present invention these insecticidal proteins or toxins are to be understood expressly also as pre- toxins, hybrid proteins, truncated or otherwise modified proteins. Hybrid proteins are characterized by a new combination of protein domains, (see, e.g. WO 02/015701 ). Further examples of such toxins or genetically modified plants capable of synthesizing such toxins are disclosed, e.g., in EP-A 374 753, WO 93/007278, WO 95/34656, EP-A 427 529, EP-A 451 878, WO 03/18810 und WO 03/52073. The methods for producing such genetically modified plants are generally known to the person skilled in the art and are described, e.g. in the publications mentioned above. These insecticidal proteins contained in the genetically modified plants impart to the plants producing these proteins tolerance to harmful pests from all taxonomic groups of arthropods, especially to beetles (Coeloptera), two-winged insects (Diptera), and moths (Lepidoptera) and to nematodes (Nematoda).

In some embodiments, expression of one or more protein toxins (e.g., insecticidal proteins) in the TK-inhibiting herbicides-tolerant plants is effective for controlling organisms that include, for example, members of the classes and orders: Coleoptera such as the American bean weevil Acanthoscelides obtectus; the leaf beetle Agelastica alni; click beetles

(Agriotes lineatus, Agriotes obscurus, Agriotes bicolor); the grain beetle Ahasverus advena; the summer schafer Amphimallon solstitialis; the furniture beetle Anobium punctatum;

Anthonomus spp. (weevils); the Pygmy mangold beetle Atomaria linearis; carpet beetles (Anthrenus spp., Attagenus spp.); the cowpea weevil Callosobruchus maculates; the fried fruit beetle Carpophilus hemipterus; the cabbage seedpod weevil Ceutorhynchus assimilis; the rape winter stem weevil Ceutorhynchus picitarsis; the wireworms Conoderus

vespertinus and Conoderus falli; the banana weevil Cosmopolites sordidus; the New

Zealand grass grub Costelytra zealandica; the June beetle Cotinis nitida; the sunflower stem weevil

Cylindrocopturus adspersus; the larder beetle Dermestes lardarius; the corn rootworms Diabrotica virgifera, Diabrotica virgifera virgifera, and Diabrotica barberi; the Mexican bean beetle Epilachna varivestis; the old house borer Hylotropes bajulus; the lucerne weevil Hypera postica; the shiny spider beetle Gibbium psylloides; the cigarette beetle Lasioderma serricorne; the Colorado potato beetle Leptinotarsa decemlineata; Lyctus beetles {Lyctus spp. , the pollen beetle Meligethes aeneus; the common cockshafer Melolontha melolontha; the American spider beetle Mezium americanum; the golden spider beetle Niptus hololeuc s; the grain beetles Oryzaephilus surinamensis and Oryzaephilus Mercator; the black vine weevil Otiorhynchus sulcatus; the mustard beetle Phaedon cochleariae, the crucifer flea beetle Phyllotreta cruciferae; the striped flea beetle Phyllotreta striolata; the cabbage steam flea beetle Psylliodes chrysocephala; Ptinus spp. (spider beetles); the lesser grain borer Rhizopertha dominica; the pea and been weevil Sitona lineatus; the rice and granary beetles Sitophilus oryzae and Sitophilus granaries; the red sunflower seed weevil

Smicronyx fulvus; the drugstore beetle Stegobium paniceum; the yellow mealworm beetle Tenebrio molitor, the flour beetles Tribolium castaneum and Tribolium confusum;

warehouse and cabinet beetles {Trogoderma spp.); the sunflower beetle Zygogramma exclamationis; Dermaptera (earwigs) such as the European earwig Forficula auricularia and the striped earwig Labidura riparia; Dictyoptera such as the oriental cockroach Blatta orientalis; the greenhouse millipede Oxidus gracilis; the beet fly Pegomyia betae; the frit fly Oscinella frit; fruitflies (Dacus spp., Drosophila spp.); Isoptera (termites) including species from the familes Hodotermitidae, Kalotermitidae, Mastotermitidae, Rhinotermitidae,

Serriterrnitidae, Termitidae, Termopsidae; the tarnished plant bug Lygus lineolahs; the black bean aphid Aphis fabae; the cotton or melon aphid Aphis gossypii; the green apple aphid Aphis pomi; the citrus spiny whitefly Aleurocanthus spiniferus; the sweet potato whitefly Bemesia tabaci; the cabbage aphid Brevicoryne brassicae; the pear psylla Cacopsylla pyricola; the currant aphid Cryptomyzus ribis; the grape phylloxera Daktulosphaira vitifoliae; the citrus psylla Diaphorina citri; the potato leafhopper Empoasca fabae; the bean leafhopper Empoasca Solana; the vine leafhopper Empoasca vitis; the woolly aphid

Eriosoma lanigerum; the European fruit scale Eulecanium corni; the mealy plum aphid

Hyalopterus arundinis; the small brown planthopper Laodelphax striatellus; the potato aphid Macrosiphum euphorbiae; the green peach aphid Myzus persicae; the green rice

leafhopper Nephotettix cinticeps; the brown planthopper Nilaparvata lugens; the hop aphid Phorodon humuli; the bird-cherry aphid Rhopalosiphum padi; the grain aphid Sitobion avenae; Lepidoptera such as Adoxophyes orana (summer fruit tortrix moth); Archips podana (fruit tree tortrix moth); Bucculatrix pyrivorella (pear leafminer); Bucculatrix thurberiella (cotton leaf perforator); Bupalus piniarius (pine looper); Carpocapsa pomonella (codling moth); Chilo suppressalis (striped rice borer); Choristoneura fumiferana (eastern spruce budworm); Cochylis hospes (banded sunflower moth); Diatraea grandiosella

(southwestern corn borer); Eupoecilia ambiguella (European grape berry moth);

Helicoverpa armigera (cotton bollworm); Helicoverpa zea (cotton bollworm); Heliothis vires cens (tobacco budworm), Homeosoma electellum (sunflower moth); Homona magnanima (oriental tea tree tortrix moth); Lithocolletis blancardella (spotted tentiform leafminer);

Lymantria dispar (gypsy moth); Malacosoma neustria (tent caterpillar); Mamestra brassicae (cabbage armyworm); Mamestra configurata (Bertha armyworm); Operophtera brumata (winter moth); Ostrinia nubilalis (European corn borer), Panolis flammea (pine beauty moth), Phyllocnistis citrella (citrus leafminer); Pieris brassicae (cabbage white butterfly); Rachiplusia ni (soybean looper); Spodoptera exigua (beet armywonn); Spodoptera littoralis (cotton leafworm); Sylepta derogata (cotton leaf roller); Trichoplusia ni (cabbage looper); Orthoptera such as the common cricket Acheta domesticus, tree locusts (Anacridium spp.), the migratory locust Locusta migratoria, the twostriped grasshopper Melanoplus bivittatus, the differential grasshopper Melanoplus differ entialis, the redlegged grasshopper

Melanoplus femurrubrum, the migratory grasshopper Melanoplus sanguinipes, the northern mole cricket Neocurtilla hexadectyla, the red locust Nomadacris septemfasciata, the shortwinged mole cricket Scapteriscus abbreviatus, the southern mole cricket Scapteriscus borellii, the tawny mole cricket Scapteriscus vicinus, and the desert locust Schistocerca gregaria; Symphyla such as the garden symphylan Scutigerella immaculata; Thysanoptera such as the tobacco thrips Frankliniella fusca, the flower thrips Frankliniella intonsa, the western flower thrips Frankliniella occidentalism the cotton bud thrips Frankliniella schultzei, the banded greenhouse thrips Hercinothrips femoralis, the soybean thrips Neohydatothrips variabilis, Kelly's citrus thrips Pezothrips kellyanus, the avocado thrips Scirtothrips perseae, the melon thrips Thrips palmi, and the onion thrips Thrips tabaci; and the like, and combinations comprising one or more of the foregoing organisms.

In some embodiments, expression of one or more protein toxins (e.g., insecticidal proteins) in the TK-inhibiting herbicides-tolerant plants is effective for controlling flea beetles, i.e. members of the flea beetle tribe of family Chrysomelidae, preferably against Phyllotreta spp., such as Phyllotreta cruciferae and/or Phyllotreta triolata. In other embodiments, expression of one or more protein toxins {e.g., insecticidal proteins) in the TK-inhibiting herbicides- tolerant plants is effective for controlling cabbage seedpod weevil, the Bertha armyworm, Lygus bugs, or the diamondback moth.

Furthermore, in one embodiment, TK-inhibiting herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, rendered able to synthesize one or more proteins to increase the resistance or tolerance of those plants to bacterial, viral or fungal pathogens. The methods for producing such genetically modified plants are generally known to the person skilled in the art. Furthermore, in another embodiment, TK-inhibiting herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, rendered able to synthesize one or more proteins to increase the productivity (e.g. oil content), tolerance to drought, salinity or other growth- limiting environmental factors or tolerance to pests and fungal, bacterial or viral pathogens of those plants.

Furthermore, in other embodiments, TK-inhibiting herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, altered to contain a modified amount of one or more substances or new substances, for example, to improve human or animal nutrition, e.g. oil crops that produce health-promoting long-chain omega-3 fatty acids or unsaturated omega-9 fatty acids (e.g. Nexera(R) rape, Dow Agro Sciences, Canada). Furthermore, in some embodiments, TK-inhibiting herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, altered to contain increased amounts of vitamins and/or minerals, and/or improved profiles of nutraceutical compounds. In one embodiment, TK-inhibiting herbicides-tolerant plants of the present invention, relative to a wild-type plant, comprise an increased amount of, or an improved profile of, a compound selected from the group consisting of: glucosinolates (e.g., glucoraphanin (4- methylsulfinylbutyl-glucosinolate), sulforaphane, 3-indolylmethyl- glucosinolate(glucobrassicin), I -methoxy-3-indolylmethyl-glucosinolate

(neoglucobrassicin)); phenolics (e.g., flavonoids (e.g., quercetin, kaempferol),

hydroxycinnamoyl derivatives (e.g., 1 ,2,2'- trisinapoylgentiobiose, 1 ,2- diferuloylgentiobiose, I ,2'-disinapoyl-2-feruloylgentiobiose, 3-0- caffeoyl-quinic

(neochlorogenic acid)); and vitamins and minerals (e.g., vitamin C, vitamin E, carotene, folic acid, niacin, riboflavin, thiamine, calcium, iron, magnesium, potassium, selenium, and zinc).

In another embodiment, TK-inhibiting herbicides-tolerant plants of the present invention, relative to a wild-type plant, comprise an increased amount of, or an improved profile of, a compound selected from the group consisting of: progoitrin; isothiocyanates; indoles (products of glucosinolate hydrolysis); glutathione; carotenoids such as beta-carotene, lycopene, and the xanthophyll carotenoids such as lutein and zeaxanthin; phenolics comprising the flavonoids such as the flavonols (e.g. quercetin, rutin), the flavans/tannins (such as the procyanidins comprising coumarin, proanthocyanidins, catechins, and anthocyanins); flavones; phytoestrogens such as coumestans, lignans, resveratrol, isoflavones e.g. genistein, daidzein, and glycitein; resorcyclic acid lactones; organosulphur compounds; phytosterols; terpenoids such as carnosol, rosmarinic acid, glycyrrhizin and saponins; chlorophyll; chlorphyllin, sugars, anthocyanins, and vanilla.

[0129] In other embodiments, TK-inhibiting herbicides-tolerant plants of the present invention, relative to a wild-type plant, comprise an increased amount of, or an improved profile of, a compound selected from the group consisting of: vincristine, vinblastine, taxanes (e.g., taxol (paclitaxel), baccatin III, 10-desacetylbaccatin III, 10-desacetyl taxol, xylosyl taxol, 7- epitaxol, 7-epibaccatin III, 10-desacetylcephalomannine, 7- epicephalomannine, taxotere, cephalomannine, xylosyl cephalomannine, taxagifine, 8- benxoyloxy taxagifine, 9-acetyloxy taxusin, 9-hydroxy taxusin, taiwanxam, taxane la, taxane lb, taxane lc, taxane Id, GMP paclitaxel, 9-dihydro 13-acetylbaccatin III, 10-desacetyl-7- epitaxol, tetrahydrocannabinol (THC), cannabidiol (CBD), genistein, diadzein, codeine, morphine, quinine, shikonin, ajmalacine, serpentine, and the like.

In other aspects, a method for treating a plant of the present invention is provided.

In some embodiments, the method comprises contacting the plant with an agronomically acceptable composition. In one embodiment, the agronomically acceptable composition comprises an auxinic herbicide A. I.

In another aspect, the present invention provides a method for preparing a descendent seed. The method comprises planting a seed of or capable of producing a plant of the present invention. In one embodiment, the method further comprises growing a descendent plant from the seed; and harvesting a descendant seed from the descendent plant. In other embodiments, the method further comprises applying a TK-inhibiting herbicides herbicidal composition to the descendent plant.

In another embodiment, the invention refers to harvestable parts of the transgenic plant according to the present invention. Preferably, the harvestable parts comprise the TK nucleic acid or TK protein of the present invention. The harvestable parts may be seeds, roots, leaves and/or flowers comprising the TK nucleic acid or TK protein or parts thereof. Preferred parts of soy plants are soy beans comprising the TK nucleic acid or TK protein. In another embodiment, the invention refers to products derived from a transgenic plant according to the present invention, parts thereof or harvestable parts thereof. A preferred plant product is fodder, seed meal, oil, or seed-treatment-coated seeds. Preferably, the meal and/or oil comprise the TK nucleic acids or TKproteins. In another embodiment, the invention refers to a method for the production of a product, which method comprises

a) growing the plants of the invention or obtainable by the methods of invention and b) producing said product from or by the plants of the invention and/or parts, e.g. seeds, of these plants.

In a further embodiment the method comprises the steps

a) growing the plants of the invention, b) removing the harvestable parts as defined above from the plants and

c) producing said product from or by the harvestable parts of the invention.

The product may be produced at the site where the plant has been grown, the plants and/or parts thereof may be removed from the site where the plants have been grown to produce the product. Typically, the plant is grown, the desired harvestable parts are removed from the plant, if feasible in repeated cycles, and the product made from the harvestable parts of the plant. The step of growing the plant may be performed only once each time the methods of the invention is performed, while allowing repeated times the steps of product production e.g. by repeated removal of harvestable parts of the plants of the invention and if necessary further processing of these parts to arrive at the product. It is also possible that the step of growing the plants of the invention is repeated and plants or harvestable parts are stored until the production of the product is then performed once for the accumulated plants or plant parts. Also, the steps of growing the plants and producing the product may be performed with an overlap in time, even simultaneously to a large extend or sequentially. Generally the plants are grown for some time before the product is produced.

In one embodiment the products produced by said methods of the invention are plant products such as, but not limited to, a foodstuff, feedstuff, a food supplement, feed supplement, fiber, cosmetic and/or pharmaceutical. Foodstuffs are regarded as

compositions used for nutrition and/or for supplementing nutrition. Animal feedstuffs and animal feed supplements, in particular, are regarded as foodstuffs.

In another embodiment the inventive methods for the production are used to make agricultural products such as, but not limited to, plant extracts, proteins, amino acids, carbohydrates, fats, oils, polymers, vitamins, and the like.

It is possible that a plant product consists of one or more agricultural products to a large extent.

Herbicides

Generally, if the TK-inhibiting herbicides (also referred to as compounds A hereinafter) and/or the herbicidal compounds B as described herein, which can be employed in the context of the present invention, are capable of forming geometrical isomers, for example E/Z isomers, it is possible to use both, the pure isomers and mixtures thereof, in the compositions useful for the present the invention. If the TK-inhibting herbicides A and/or the herbicidal compounds B as described herein have one or more centers of chirality and, as a consequence, are present as enantiomers or diastereomers, it is possible to use both, the pure enantiomers and diastereomers and their mixtures, in the compositions according to the invention. If the TK-inhibting herbicides A and/or the herbicidal compounds B as described herein have ionizable functional groups, they can also be employed in the form of their agriculturally acceptable salts. Suitable are, in general, the salts of those cations and the acid addition salts of those acids whose cations and anions, respectively, have no adverse effect on the activity of the active compounds. Preferred cations are the ions of the alkali metals, preferably of lithium, sodium and potassium, of the alkaline earth metals, preferably of calcium and magnesium, and of the transition metals, preferably of

manganese, copper, zinc and iron, further ammonium and substituted ammonium in which one to four hydrogen atoms are replaced by Ci-C₄-alkyl, hydroxy-Ci-C₄-alkyl, Ci-C₄-alkoxy- Ci-C₄-alkyl, hydroxy-Ci-C₄-alkoxy-Ci-C₄-alkyl, phenyl or benzyl, preferably ammonium, methylammonium, isopropylammonium, dimethylammonium, diisopropylammonium, trimethylammonium, heptylammonium, dodecylammonium, tetradecylammonium, tetramethylammonium, tetraethylammonium, tetrabutylammonium, 2-hydroxyethyl- ammonium (olamine salt), 2-(2-hydroxyeth-1-oxy)eth-1 -ylammonium (diglycolamine salt), di(2-hydroxyeth-1 -yl)ammonium (diolamine salt), tris(2-hydroxyethyl)ammonium (trolamine salt), tris(2-hydroxypropyl)ammonium, benzyltrimethylammonium, benzyltriethylammonium, Ν,Ν,Ν-trimethylethanolammonium (choline salt), furthermore phosphonium ions, sulfonium ions, preferably tri(Ci-C₄-alkyl)sulfonium, such as trimethylsulfonium, and sulfoxonium ions, preferably tri(Ci-C₄-alkyl)sulfoxonium, and finally the salts of polybasic amines such as N,N- bis-(3-aminopropyl)methylamine and diethylenetriamine. Anions of useful acid addition salts are primarily chloride, bromide, fluoride, iodide, hydrogensulfate, methylsulfate, sulfate, dihydrogenphosphate, hydrogenphosphate, nitrate, bicarbonate, carbonate,

hexafluorosilicate, hexafluorophosphate, benzoate and also the anions of Ci-C₄-alkanoic acids, preferably formate, acetate, propionate and butyrate.

The TK-inhibting herbicides A and/or the herbicidal compounds B as described herein having a carboxyl group can be employed in the form of the acid, in the form of an agriculturally suitable salt as mentioned above or else in the form of an agriculturally acceptable derivative, for example as amides, such as mono- and di-Ci-C6-alkylamides or arylamides, as esters, for example as allyl esters, propargyl esters, Ci-Cio-alkyl esters, alkoxyalkyl esters, tefuryl ((tetrahydrofuran-2-yl)methyl) esters and also as thioesters, for example as Ci-Cio-alkylthio esters. Preferred mono- and di-Ci-C6-alkylamides are the methyl and the dimethylamides. Preferred arylamides are, for example, the anilides and the 2-chloroanilides. Preferred alkyl esters are, for example, the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, mexyl (1 -methylhexyl), meptyl (1 -methylheptyl), heptyl, octyl or isooctyl (2-ethylhexyl) esters. Preferred Ci-C₄-alkoxy-Ci-C₄-alkyl esters are the straight- chain or branched Ci-C₄-alkoxy ethyl esters, for example the 2-methoxyethyl, 2-ethoxyethyl, 2-butoxyethyl (butotyl), 2-butoxypropyl or 3-butoxypropyl ester. An example of a straight- chain or branched Ci-Cio-alkylthio ester is the ethylthio ester.

Examples of TK inhibiting herbicides which can be used according to the present invention are compounds having the Formula I.a1 , I.a2, I.a3, or I.a4, known to the skilled artisan as (Hydroxy- ) Cornexisthin. Cornexistin (I.a1 ) and its corresponding dibasic acid (I.a2) are known from EP-A 0 290 193:

Hydroxycornexistin (I.a3) and its corresponding dibasic acid (I.a4) are known from US

5,424,278:

The herbicidal activity of these compounds is also known from the cited references. JP-A 1990- 256 602 discloses mixtures of cornexistin (I.a1 ) and its dibasic acid (I.a2) with certain herbicides.

Nevertheless, there is still room for improvement, e.g. regarding activity, scope of activity and compatibility with useful plants of the known herbicidal compositions.

It is an object of the present invention to provide methods of using herbicidal compositions which are highly active against unwanted harmful plants. At the same time, the compositions should have good compatibility with the plants of the present invention. In addition, the compositions useful for the invention should have a broad spectrum of activity. A further object of the present invention is reducing the application rates of active ingredients.

This and further objects are achieved by the herbicidal compositions below.

Accordingly, in one aspect of the invention the TK-inhibiting herbicide usefuld for the present invention refers to a herbicidal composition comprising:

A) at least one herbicidally active compound of formula I (herbicide A)

wherein

R¹ is CH₃ or CH₂OH and

R² and R³ together with the neighbouring carbon atoms form a dihydro-2,5-dioxofuran ring or

including agriculturally acceptable salts and derivatives thereof; The TK-inhibiting herbicides described above that are useful to carry out the present invention are often best applied in conjunction with one or more other herbicides to obtain control of a wider variety of undesirable vegetation. For example, TK-inhibiting herbicides may further be used in conjunction with additional herbicides to which the crop plant is naturally tolerant, or to which it is resistant via expression of one or more additional transgenes as mentioned supra. When used in conjunction with other targeting herbicides, the TK-inhibiting herbicides, to which the plant of the present invention had been made resistant or tolerant, can be formulated with the other herbicide or herbicides, tank mixed with the other herbicide or herbicides, or applied sequentially with the other herbicide or herbicides.

Suitable components for mixtures are, for example, selected from the herbicides of class b1 ) to b15)

B) herbicides of class b1 ) to b15):

b1 ) lipid biosynthesis inhibitors;

b2) acetolactate synthase inhibitors (ALS inhibitors);

b3) photosynthesis inhibitors;

b4) protoporphyrinogen-IX oxidase inhibitors,

b5) bleacher herbicides;

b6) enolpyruvyl shikimate 3-phosphate synthase inhibitors (EPSP inhibitors); b7) glutamine synthetase inhibitors;

b8) 7,8-dihydropteroate synthase inhibitors (DHP inhibitors);

b9) mitosis inhibitors;

b10) inhibitors of the synthesis of very long chain fatty acids (VLCFA inhibitors); b1 1 ) cellulose biosynthesis inhibitors;

b12) decoupler herbicides;

b13) auxinic herbicides;

b14) auxin transport inhibitors; and

b15) other herbicides selected from the group consisting of bromobutide,

chlorflurenol, chlorflurenol-methyl, cinmethylin, cumyluron, dalapon, dazomet, difenzoquat, difenzoquat-metilsulfate, dimethipin, DSMA, dymron, endothal and its salts, etobenzanid, flamprop, flamprop-isopropyl, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl, flurenol, flurenol-butyl, flurprimidol, fosamine, fosamine-ammonium, indanofan, indaziflam, maleic hydrazide, mefluidide, metam, methiozolin (CAS 403640-27-7), methyl azide, methyl bromide, methyl-dymron, methyl iodide, MSMA, oleic acid, oxaziclomefone, pelargonic acid, pyributicarb, quinoclamine, triaziflam, tridiphane and 6-chloro- 3-(2-cyclopropyl-6-methylphenoxy)-4-pyridazinol (CAS 499223-49-3) and its salts and esters; including their agriculturally acceptable salts or derivatives.

The compositions according to the invention are suitable as herbicides as such or as

appropriately formulated compositions (agrochemical compositions). As used herein, the term "agrochemical composition" refers to a composition comprising a pesticidally effective amount of at least one active ingredient and at least one auxiliary customary for agrochemical

compositions. The invention relates in particular to the use of compositions in the form of herbicidally active agrochemical compositions comprising a herbicidally effective amount of A) at least one compound of formula I (herbicide A) and B) at least one further compound selected from the herbicides of groups b1 ) to b15) (herbicide B), as defined above, and also at least one liquid and/or solid carrier and/or one or more surfactants and, if desired, one or more further auxiliaries customary for agrochemical compositions.

The invention also relates to compositions in the form of an agrochemical composition, which is a 1 -component composition comprising at least one herbicide A and at least one further active compound selected from the herbicides B, and at least one solid or liquid carrier and/or one or more surfactants and, if desired, one or more further auxiliaries customary for agrochemical compositions.

The invention also relates to compositions in the form of an agrochemical composition, which is a 2-component composition comprising a first component comprising at least one herbicide A, a solid or liquid carrier and/or one or more surfactants, and a second component comprising at least one herbicide B, a solid or liquid carrier and/or one or more surfactants, where additionally both components may also comprise further auxiliaries customary for agrochemical

compositions. Moreover, the time frame, within which the desired herbicidal action can be achieved, may be expanded by the compositions according to the invention comprising at least one herbicide A and at least one herbicide B and optionally a safener C as defined below. This allows a more flexibly timed application of the compositions according to the invention in comparison with the single compounds.

Safeners are chemical compounds which prevent or reduce damage on useful plants without having a major impact on the herbicidal action of the herbicidal active components towards unwanted plants. Safeners can be applied before sowings (e.g. seed treatments), on shoots or seedlings as well as in the pre-emergence or post-emergence treatment of useful plants and their habitat.

The compositions according to the invention comprising both at least one herbicide A and at least one safener C as defined below also have good herbicidal activity against harmful plants and better compatibility with useful plants.

Surprisingly, compositions according to the invention comprising at least one herbicide A at least one herbicide B and at least one safener C have better herbicidal activity, i.e. better activity against harmful plants, than would have been expected based on the herbicidal activity observed for the individual compounds, or a broader activity spectrum, and show better compatibility with useful plants than compositions comprising only one herbicide A and one herbicide B.

Therefore, in one embodiment of the present invention the compositions comprise at least one herbicide A, at least one herbicide B and at least one safener C.

Examples of suitable safeners C are benoxacor, cloquintocet, cyometrinil, cyprosulfamide, dichlormid, dicyclonon, dietholate, fenchlorazole, fenclorim, flurazole, fluxofenim, furilazole, isoxadifen, mefenpyr, mephenate, naphthalic anhydride, oxabetrinil, 4-(dichloroacetyl)-1 -oxa-4- azaspiro[4.5]decane (MON4660, CAS 71526-07-3), 2,2,5-trimethyl-3-(dichloroacetyl)-1 ,3- oxazolidine (R-29148, CAS 52836-31 -4) and N-(2-methoxybenzoyl)-4-[(methylamino- carbonyl)amino]benzenesulfonamide (CAS 129531 -12-0).

The safeners C, the herbicides A and the herbicides B can be applied simultaneously or in succession.

The term "cornexistin" means the compound of formula (I.a1 ) as well as agriculturally acceptable salts thereof. The term "dibasic acid of cornexistin" means the compound of formula (I.a2) as well as agriculturally acceptable salts thereof.

The term "hydroxycornexistin" means the compound of formula (I.a3) as well as agriculturally acceptable salts thereof.

The term "dibasic acid of hydroxycornexistin" means the compound of formula (I.a4) as well as agriculturally acceptable salts thereof.

The compounds of formulae (I .a 1 ) to (I.a4) as described herein are capable of forming geometrical isomers, for example E/Z isomers. Accordingly, the terms "cornexistin", "dibasic acid of cornexistin", "hydroxycornexistin" and "dibasic acid of hydroxycornexistin" also encompass the pure E or Z isomers and mixtures thereof.

The term "agriculturally acceptable salts" is used herein to mean in general, the salts of those cations and the acid addition salts of those acids whose cations and anions, respectively, have no adverse effect on the herbicidal activity of the dibasic acid of cornexistin and the dibasic acid of hydroxycornexistin.

Preferred cations are the ions of the alkali metals, preferably of lithium, sodium and potassium, of the alkaline earth metals, preferably of calcium and magnesium, and of the transition metals, preferably of manganese, copper, zinc and iron, further ammonium and substituted ammonium in which one to four hydrogen atoms are replaced by Ci-C4-alkyl, hydroxy-Ci-C4-alkyl, C1-C4- alkoxy-Ci-C4-alkyl, hydroxy-Ci-C4-alkoxy-Ci-C4-alkyl, phenyl or benzyl, preferably ammonium, methylammonium, isopropylammonium, dimethylammonium, diisopropylammonium, trimethylammonium, heptylammonium, dodecylammonium, tetradecylammonium,

tetramethylammonium, tetraethylammonium, tetrabutylammonium, 2 hydroxyethyl-ammonium (olamine salt), 2-(2-hydroxyeth-1 -oxy)eth-1 -ylammonium (diglycolamine salt), di(2-hydroxyeth-1 - yl)-ammonium (diolamine salt), tris(2-hydroxyethyl)ammonium (trolamine salt), tris(2-hydroxy- propyl)ammonium, benzyltrimethylammonium, benzyltriethylammonium, Ν,Ν,Ν-trimethylethanol- ammonium (choline salt), furthermore phosphonium ions, sulfonium ions, preferably tri(Ci-C4- alkyl)sulfonium, such as trimethylsulfonium, and sulfoxonium ions, preferably tri(Ci-C4- alkyl)sulfoxonium, and finally the salts of polybasic amines such as N,N-bis-(3-amino- propyl)methylamine and diethylenetriamine.

Anions of useful acid addition salts are primarily chloride, bromide, fluoride, iodide,

hydrogensulfate, methylsulfate, sulfate, dihydrogenphosphate, hydrogen-'phosphate, nitrate, bicarbonate, carbonate, hexafluorosilicate, hexafluorophosphate, benzoate and also the anions of Ci-C4-alkanoic acids, preferably formate, acetate, propionate and butyrate.

If the herbicidal compounds B and/or D and/or safeners C as described herein are capable of forming geometrical isomers, for example E/Z isomers, it is possible to use both, the pure isomers and mixtures thereof, in the compositions according to the invention. If the herbicidal compounds B and/or D and/or safeners C as described herein have one or more centers of chirality and, as a consequence, are present as enantiomers or diastereomers, it is possible to use both, the pure enantiomers and diastereomers and their mixtures, in the compositions according to the invention. If the herbicidal compounds B and/or D and/or safeners C as described herein have ionizable functional groups, they can also be employed in the form of their agriculturally acceptable salts. Suitable are, in general, the salts of those cations and the acid addition salts of those acids whose cations and anions, respectively, have no adverse effect on the activity of the active compounds.

Preferred cations and anions are the ones listed for the compounds of formula I.

Herbicidal compounds A, B and/or D and/or safeners C as described herein having a carboxyl, hydroxy and/or an amino group can be employed as such or in form of an agriculturally suitable salt as mentioned above or else in the form of an agriculturally acceptable derivative, for example as amides, such as mono- and di-Ci-C6-alkylamides or arylamides, as esters, for example as allyl esters, propargyl esters, Ci-Cio-alkyl esters, alkoxyalkyl esters, tefuryl ((tetrahydrofuran-2-yl)methyl) esters and also as thioesters, for example as Ci-Cio-alkylthio esters. Preferred mono- and di-Ci-C6-alkylamides are the methyl and the dimethylamides. Preferred arylamides are, for example, the anilides and the 2-chloroanilides. Preferred alkyl esters are, for example, the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, mexyl (1 - methylhexyl), meptyl (1 -methylheptyl), heptyl, octyl or isooctyl (2-ethylhexyl) esters. Preferred Ci-C4-alkoxy-Ci-C4-alkyl esters are the straight-chain or branched Ci-C4-alkoxy ethyl esters, for example the 2-methoxyethyl, 2-ethoxyethyl, 2-butoxyethyl (butotyl), 2-butoxypropyl or 3- butoxypropyl ester. An example of a straight-chain or branched Ci-Cio-alkylthio ester is the ethylthio ester.

Further embodiments of the invention are evident from the claims, the description and the examples. It is to be understood that the features mentioned above and still to be illustrated below of the subject matter of the invention can be applied not only in the combination given in each particular case but also in other combinations, without leaving the scope of the invention.

The preferred embodiments of the invention mentioned herein below have to be understood as being preferred either independently from each other or in combination with one another.

Preferably, the active ingredient A is selected from the following herbicides A: cornexistin (I.a1 ),

the dibasic acid of cornexistin (I.a2),

hydroxycornexistin (I.a3),

the dibasic acid of hydroxycornexistin (I.a4),

mixtures of cornexistin (I .a 1 ) and the dibasic acid of cornexistin (I.a2),

mixtures of hydroxycornexistin (I.a3) and the dibasic acid of hydroxycornexistin (I.a4) mixtures of cornexistin (I .a 1 ) and hydroxycornexistin (I.a3),

mixtures of cornexistin (I.a1 ) and the dibasic acid of hydroxycornexistin (I.a4),

mixtures of the dibasic acid of cornexistin (I.a2) and hydroxycornexistin (I.a3),

a10) mixtures of the dibasic acid of cornexistin (I.a2) and the dibasic acid of hydroxycornexistin (I.a4) and a1 1 ) mixtures cornexistin (I.a1 ), the dibasic acid of cornexistin (I.a2), hydroxycornexistin (I.a3) and the dibasic acid of hydroxycornexistin (I.a4) including agriculturally acceptable salts and derivatives thereof.

Prefered are the compositions according to the present invention comprising at least one, preferably exactly one herbicide A and at least one herbicide B.

Further preferred are compositions according to the present invention comprising at least two, preferably exactly two herbicides A and at least one herbicide B.

In one preferred embodiment of the invention the compositions contain at least one inhibitor of the lipid biosynthesis (herbicide b1 ) through inhibition of acetylCoA carboxylase (hereinafter termed ACC herbicides). The ACC herbicides belong to the group A of the HRAC classification system.

According to a further embodiment of the invention the compositions contain at least one inhibitor of photosynthesis (herbicide b3) on diverting the electron transfer in photosystem I in plants (so-called PSI inhibitors, group D of HRAC classification) and thus on an inhibition of photosynthesis.

According to a further embodiment of the invention the compositions contain at least one inhibitor of protoporphyrinogen-IX-oxidase (herbicide b4). The herbicidal activity of these compounds is based on the inhibition of the protoporphyrinogen-IX-oxidase. These inhibitors belong to the group E of the HRAC classification system.

According to a further embodiment of the invention the compositions contain at least one bleacher-herbicide (herbicide b5). The herbicidal activity of these compounds is based on the inhibition of the carotenoid biosynthesis. These include compounds which inhibit carotenoid biosynthesis by inhibition of phytoene desaturase (so-called PDS inhibitors, group F1 of HRAC classification), compounds that inhibit the 4-hydroxyphenylpyruvate-dioxygenase (HPPD inhibitors, group F2 of HRAC classification), compounds that inhibit DOXsynthase (group F4 of HRAC class) and compounds which inhibit carotenoid biosynthesis by an unknown mode of action (bleacher - unknown target, group F3 of HRAC classification).

According to a further embodiment of the invention the compositions contain at least one EPSP synthase inhibitor (herbicide b6). The herbicidal activity of these compounds is based on the inhibition of enolpyruvyl shikimate 3-phosphate synthase, and thus on the inhibition of the amino acid biosynthesis in plants. These inhibitors belong to the group G of the HRAC classification system.

According to a further embodiment of the invention the compositions contain at least one glutamine synthetase inhibitor (herbicide b7). The herbicidal activity of these compounds is based on the inhibition of glutamine synthetase, and thus on the inhibition of the aminoacid biosynthesis in plants. These inhibitors belong to the group H of the HRAC classification system. According to a further embodiment of the invention the compositions contain at least one DHP synthase inhibitor (herbicide b8). The herbicidal activity of these compounds is based on the inhibition of 7,8-dihydropteroate synthase. These inhibitors belong to the group I of the HRAC classification system. According to a further embodiment of the invention the compositions contain at least one cellulose biosynthesis inhibitor (herbicide b1 1 ). The herbicidal activity of these compounds is based on the inhibition of the biosynthesis of cellulose and thus on the inhibition of the synthesis of cell walls in plants. These inhibitors belong to the group L of the HRAC classification system. According to a further embodiment of the invention the compositions contain at least one decoupler herbicide (herbicide b12). The herbicidal activity of these compounds is based on the disruption of the cell membrane. These inhibitors belong to the group M of the HRAC classification system. According to a further embodiment of the invention the compositions contain at least one auxinic herbicide (herbicide b13). These include compounds that mimic auxins, i.e. plant hormones, and affect the growth of the plants. These compounds belong to the group O of the HRAC classification system. According to a further embodiment of the invention the compositions contain at least one auxin transport inhibitor (herbicide b14). The herbicidal activity of these compounds is based on the inhibition of the auxin transport in plants. These compounds belong to the group P of the HRAC classification system. According to a further embodiment of the invention the compositions contain at least one other herbicide (herbicide b15) selected from the group consisting of bromobutide, chlorflurenol, chlorflurenol-methyl, cinmethylin, cumyluron, dalapon, dazomet, difenzoquat, difenzoquat- metilsulfate, dimethipin, DSMA, dymron, endothal and its salts, etobenzanid, flamprop, flamprop-isopropyl, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl, flurenol, flurenol-butyl, flurprimidol, fosamine, fosamine-ammonium, indanofan, maleic hydrazide, mefluidide, metam, methiozolin (CAS 403640-27-7), methyl azide, methyl bromide, methyl- daimuron, methyl iodide, MSMA, oleic acid, oxaziclomefone, pelargonic acid, pyributicarb, quinoclamine, and 6-chloro-3-(2-cyclopropyl-6-methylphenoxy)-4-pyridazinol (CAS 499223-49- 3). These compounds have an unknown mode of action and belong to the group Z of the HRAC classification system.

As to the given mechanisms of action and classification of the active compounds, see e.g. "HRAC, Classification of Herbicides According to Mode of Action", (http://www.plantprotection.org/hrac/MOA.html).

Examples of herbicides B which can be used in combination with the TK-inhibiting herbicides according to the present invention are: b1) from the group of the lipid biosynthesis inhibitors:

ACC-herbicides such as alloxydim, alloxydim-sodium, butroxydim, clethodim, clodinafop, clodinafop-propargyl, cycloxydim, cyhalofop, cyhalofop-butyl, diclofop, diclofop-methyl, fenoxaprop, fenoxaprop-ethyl, fenoxaprop-P, fenoxaprop-P-ethyl, fluazifop, fluazifop-butyl, fluazifop-P, fluazifop-P-butyl, haloxyfop, haloxyfop-methyl, haloxyfop-P, haloxyfop-P-methyl, metamifop, pinoxaden, profoxydim, propaquizafop, quizalofop, quizalofop-ethyl, quizalofop- tefuryl, quizalofop-P, quizalofop-P-ethyl, quizalofop-P-tefuryl, sethoxydim, tepraloxydim, tralkoxydim,

4-(4'-Chloro-4-cyclopropyl-2'-fluoro[1 ,1 '-biphenyl]-3-yl)-5-hydroxy-2,2,6,6-tetramethyl-2H- pyran-3(6H)-one (CAS 1312337-72-6); 4-(2^,,4^,-Dichloro-4-cyclopropyl[1 ,1 ^,-biphenyl]-3-yl)-5- hydroxy-2,2,6,6-tetramethyl-2H-pyran-3(6H)-one (CAS 1312337-45-3); 4-(4'-Chloro-4-ethyl- 2'-fluoro[1 ,1 '-biphenyl]-3-yl)-5-hydroxy-2,2,6,6-tetramethyl-2H-pyran-3(6H)-one (CAS 1033757-93-5); 4-(2^,,4^,-Dichloro-4-ethyl[1 ,1 ^,-biphenyl]-3-yl)-2,2,6,6-tetramethyl-2H-pyran- 3,5(4H,6H)-dione (CAS 1312340-84-3); 5-(Acetyloxy)-4-(4'-chloro-4-cyclopropyl-2'- fluoro[1 ,1 '-biphenyl]-3-yl)-3,6-dihydro-2,2,6,6-tetramethyl-2H-pyran-3-one (CAS 1312337- 48-6); 5-(Acetyloxy)-4-(2^',4'-dichloro-4-cyclopropyl- [1 ,1 '-biphenyl]-3-yl)-3,6-dihydro-2,2,6,6- tetramethyl-2H-pyran-3-one; 5-(Acetyloxy)-4-(4'-chloro-4-ethyl-2'-fluoro[1 ,1 '-biphenyl]-3-yl)- 3,6-dihydro-2,2,6,6-tetramethyl-2H-pyran-3-one (CAS 1312340-82-1 ); 5-(Acetyloxy)-4-(2',4'- dichloro-4-ethyl[1 ,1 '-biphenyl]-3-yl)-3,6-dihydro-2,2,6,6-tetramethyl-2H-pyran-3-one (CAS 1033760-55-2); 4-(4^,-Chloro-4-cyclopropyl-2^,-fluoro[1 ,1 ^,-biphenyl]-3-yl)-5,6-dihydro-2,2,6,6- tetramethyl-5-oxo-2H-pyran-3-yl carbonic acid methyl ester (CAS 1312337-51 -1 ); 4-(2^',4'- Dichloro -4-cyclopropyl- [1 ,1 '-biphenyl]-3-yl)-5,6-dihydro-2,2,6,6-tetramethyl-5-oxo-2H- pyran-3-yl carbonic acid methyl ester; 4-(4'-Chloro-4-ethyl-2'-fluoro[1 ,1 '-biphenyl]-3-yl)-5,6- dihydro-2,2,6,6-tetramethyl-5-oxo-2H-pyran-3-yl carbonic acid methyl ester (CAS 1312340- 83-2); 4-(2\4^,-Dichloro^-ethyl[1 ,1 ^,-biphenyl]-3-yl)-5,6-dihydro-2,2,6,6-tetramethyl-5-oxo-2H- pyran-3-yl carbonic acid methyl ester (CAS 1033760-58-5); and non ACC herbicides such as benfuresate, butylate, cycloate, dalapon, dimepiperate, EPTC, esprocarb, ethofumesate, flupropanate, molinate, orbencarb, pebulate, prosulfocarb, TCA, thiobencarb, tiocarbazil, triallate and vernolate; b2) from the group of the ALS inhibitors:

sulfonylureas such as amidosulfuron, azimsulfuron, bensulfuron, bensulfuron-methyl, chlorimuron, chlorimuron-ethyl, chlorsulfuron, cinosulfuron, cyclosulfamuron,

ethametsulfuron, ethametsulfuron-methyl, ethoxysulfuron, flazasulfuron, flucetosulfuron, flupyrsulfuron, flupyrsulfuron-methyl-sodium, foramsulfuron, halosulfuron, halosulfuron- methyl, imazosulfuron, iodosulfuron, iodosulfuron-methyl-sodium, iofensulfuron,

iofensulfuron-sodium, mesosulfuron, metazosulfuron, metsulfuron, metsulfuron-methyl, nicosulfuron, orthosulfamuron, oxasulfuron, primisulfuron, primisulfuron-methyl, propyrisulfuron, prosulfuron, pyrazosulfuron, pyrazosulfuron-ethyl, rimsulfuron,

sulfometuron, sulfometuron-methyl, sulfosulfuron, thifensulfuron, thifensulfuron-methyl, triasulfuron, tribenuron, tribenuron-methyl, trifloxysulfuron, triflusulfuron, triflusulfuron-methyl and tritosulfuron,

imidazolinones such as imazamethabenz, imazamethabenz-methyl, imazamox, imazapic, imazapyr, imazaquin and imazethapyr, triazolopyrimidine herbicides and sulfonanilides such as cloransulam, cloransulam-methyl, diclosulam, flumetsulam, florasulam, metosulam, penoxsulam, pyrimisulfan and pyroxsulam,

pyrimidinylbenzoates such as bispyribac, bispyribac-sodium, pyribenzoxim, pyriftalid, pyriminobac, pyriminobac-methyl, pyrithiobac, pyrithiobac-sodium, 4-[[[2-[(4,6-dimethoxy-2- pyrimidinyl)oxy]phenyl]methyl]amino]-benzoic acid-1 -methylethyl ester (CAS 420138-41 -6), 4-[[[2-[(4,6-dimethoxy-2-pyrimidinyl)oxy]phenyl]methyl]amino]-benzoic acid propyl ester (CAS 420138-40-5), N-(4-bromophenyl)-2-[(4,6-dimethoxy-2- pyrimidinyl)oxy]benzenemethanamine (CAS 420138-01 -8),

sulfonylaminocarbonyl-triazolinone herbicides such as flucarbazone, flucarbazone-sodium, propoxycarbazone, propoxycarbazone-sodium, thiencarbazone and thiencarbazone-methyl; and triafamone;

among these, a preferred embodiment of the invention relates to those compositions comprising at least one imidazolinone herbicide; b3) from the group of the photosynthesis inhibitors:

amicarbazone, inhibitors of the photosystem II, e.g. triazine herbicides, including of chlorotriazine, triazinones, triazindiones, methylthiotriazines and pyridazinones such as ametryn, atrazine, chloridazone, cyanazine, desmetryn, dimethametryn,hexazinone, metribuzin, prometon, prometryn, propazine, simazine, simetryn, terbumeton, terbuthylazin, terbutryn and trietazin, aryl urea such as chlorobromuron, chlorotoluron, chloroxuron, dimefuron, diuron, fluometuron, isoproturon, isouron, linuron, metamitron,

methabenzthiazuron, metobenzuron, metoxuron, monolinuron, neburon, siduron, tebuthiuron and thiadiazuron, phenyl carbamates such as desmedipham, karbutilat, phenmedipham, phenmedipham-ethyl, nitrile herbicides such as bromofenoxim, bromoxynil and its salts and esters, ioxynil and its salts and esters, uraciles such as bromacil, lenacil and terbacil, and bentazon and bentazon-sodium, pyridate, pyridafol, pentanochlor and propanil and inhibitors of the photosystem I such as diquat, diquat-dibromide, paraquat, paraquat-dichloride and paraquat-dimetilsulfate. Among these, a preferred embodiment of the invention relates to those compositions comprising at least one aryl urea herbicide. Among these, likewise a preferred embodiment of the invention relates to those

compositions comprising at least one triazine herbicide. Among these, likewise a preferred embodiment of the invention relates to those compositions comprising at least one nitrile herbicide; b4) from the group of the protoporphyrinogen-IX oxidase inhibitors: acifluorfen, acifluorfen-sodium, azafenidin, bencarbazone, benzfendizone, bifenox, butafenacil, carfentrazone, carfentrazone-ethyl, chlomethoxyfen, cinidon-ethyl, fluazolate, flufenpyr, flufenpyr-ethyl, flumiclorac, flumiclorac-pentyl, flumioxazin, fluoroglycofen, fluoroglycofen-ethyl, fluthiacet, fluthiacet-methyl, fomesafen, halosafen, lactofen, oxadiargyl, oxadiazon, oxyfluorfen, pentoxazone, profluazol, pyraclonil, pyraflufen, pyraflufen-ethyl, saflufenacil, sulfentrazone, thidiazimin, tiafenacil, ethyl [3-[2-chloro-4-fluoro-5-(1 -methyl-6- trifluoromethyl-2,4-dioxo-1 ,2,3,4-tetrahydropyrimidin-3-yl)phenoxy]-2-pyridyloxy]acetate (CAS 353292-31 -6; S-3100, N-ethyl-3-(2,6-dichloro-4-trifluoromethylphenoxy)-5-methyl-1 H- pyrazole-1 -carboxamide (CAS 452098-92-9), N-tetrahydrofurfuryl-3-(2,6-dichloro-4- trifluoromethylphenoxy)-5-methyl-1 H-pyrazole-1 -carboxamide (CAS 915396-43-9), N-ethyl- 3-(2-chloro-6-fluoro-4-trifluoromethylphenoxy)-5-methyl-1 H-pyrazole-1 -carboxamide (CAS 452099-05-7), N-tetrahydrofurfuryl-3-(2-chloro-6-fluoro-4-trifluoromethylphenoxy)-5-methyl- 1 H-pyrazole-1 -carboxamide (CAS 452100-03-7), 3-[7-fluoro-3-oxo-4-(prop-2-ynyl)-3,4- dihydro-2H-benzo[1 ,4]oxazin-6-yl]-1 ,5-dimethyl-6-thioxo-[1 ,3,5]triazinan-2,4-dione, 1 ,5- dimethyl-6-thioxo-3-(2,2,7-trifluoro-3-oxo-4-(prop-2-ynyl)-3,4-dihydro-2H- benzo[b][1 ,4]oxazin-6-yl)-1 ,3,5-triazinane-2,4-dione (CAS 1258836-72-4), 2-(2,2,7-Trifluoro- 3-oxo-4-prop-2-ynyl-3,4-dihydro-2H-benzo[1 ,4]oxazin-6-yl)-4,5,6,7-tetrahydro-isoindole-1 ,3- dione, 1 -Methyl-6-trifluoromethyl-3-(2,2,7-trifluoro-3-oxo-4-prop-2-ynyl-3,4-dihydro-2H- benzo[1 ,4]oxazin-6-yl)-1 H-pyrimidine-2,4-dione (CAS 1304113-05-0), methyl (£)-4-[2- chloro-5-[4-chloro-5-(difluoromethoxy)-1 H-methyl-pyrazol-3-yl]-4-fluoro-phenoxy]-3- methoxy-but-2-enoate [CAS 948893-00-37, and 3-[7-Chloro-5-fluoro-2-(trifluoromethyl)-1 H- benzimidazol-4-yl]-1 -methyl-6-(trifluoromethyl)-1 H-pyrimidine-2,4-dione (CAS 212754-02-4); b5) from the group of the bleacher herbicides:

PDS inhibitors: beflubutamid, diflufenican, fluridone, flurochloridone, flurtamone,

norflurazon, picolinafen, and 4-(3-trifluoromethylphenoxy)-2-(4-trifluoromethylphenyl)- pyrimidine (CAS 180608-33-7), HPPD inhibitors: benzobicyclon, benzofenap, clomazone, isoxaflutole, mesotrione, pyrasulfotole, pyrazolynate, pyrazoxyfen, sulcotrione, tefuryltrione, tembotrione, topramezone and bicyclopyrone, bleacher, unknown target: aclonifen, amitrole and flumeturon; b6) from the group of the EPSP synthase inhibitors:

glyphosate, glyphosate-isopropylammonium, glyposate-potassium and glyphosate- trimesium (sulfosate); b7) from the group of the glutamine synthase inhibitors:

bilanaphos (bialaphos), bilanaphos-sodium, glufosinate, glufosinate-P and glufosinate- ammonium; b8) from the group of the DHP synthase inhibitors:

asulam; b9) from the group of the mitosis inhibitors:

compounds of group K1 : dinitroanilines such as benfluralin, butralin, dinitramine,

ethalfluralin, fluchloralin, oryzalin, pendimethalin, prodiamine and trifluralin,

phosphoramidates such as amiprophos, amiprophos-methyl, and butamiphos, benzoic acid herbicides such as chlorthal, chlorthal-dimethyl, pyridines such as dithiopyr and thiazopyr, benzamides such as propyzamide and tebutam; compounds of group K2: chlorpropham, propham and carbetamide, among these, compounds of group K1 , in particular

dinitroanilines are preferred; b10) from the group of the VLCFA inhibitors:

chloroacetamides such as acetochlor, alachlor, butachlor, dimethachlor, dimethenamid, dimethenamid-P, metazachlor, metolachlor, metolachlor-S, pethoxamid, pretilachlor, propachlor, propisochlor and thenylchlor, oxyacetanilides such as flufenacet and

mefenacet, acetanilides such as diphenamid, naproanilide and napropamide, tetrazolinones such fentrazamide, and other herbicides such as anilofos, cafenstrole, fenoxasulfone, ipfencarbazone, piperophos, pyroxasulfone and isoxazoline compounds of the formulae 11.1 , II.2, 11.3, II.4, II.5, II.6, II.7, 11.8 and II.9

II.6 I.7

11.9

11.8

the isoxazoline compounds of the formula (1)1 are known in the art, e.g. from WO

2006/024820, WO 2006/037945, WO 2007/071900 and WO 2007/096576; among the VLCFA inhibitors, preference is given to chloroacetamides and oxyacetamides; b11 ) from the group of the cellulose biosynthesis inhibitors:

chlorthiamid, dichlobenil, flupoxam, indaziflam, triaziflam, isoxaben and 1 -Cyclohexyl-5- pentafluorphenyloxy-1⁴-[1 ,2,4,6]thiatriazin-3-ylamine; b12) from the group of the decoupler herbicides:

dinoseb, dinoterb and DNOC and its salts; b13) from the group of the auxinic herbicides:

2,4-D and its salts and esters such as clacyfos, 2,4-DB and its salts and esters,

aminocyclopyrachlor and its salts and esters, aminopyralid and its salts such as

aminopyralid-tris(2-hydroxypropyl)ammonium and its esters, benazolin, benazolin-ethyl, chloramben and its salts and esters, clomeprop, clopyralid and its salts and esters, dicamba and its salts and esters, dichlorprop and its salts and esters, dichlorprop-P and its salts and esters, fluroxypyr, fluroxypyr-butometyl, fluroxypyr-meptyl, halauxifen and its salts and esters (CAS 943832-60-8); MCPA and its salts and esters, MCPA-thioethyl, MCPB and its salts and esters, MCPP and its salts and esters, mecoprop and its salts and esters, mecoprop-P and its salts and esters, picloram and its salts and esters, quinclorac, quinmerac, TBA (2,3,6) and its salts and esters and triclopyr and its salts and esters; b14) from the group of the auxin transport inhibitors: diflufenzopyr, diflufenzopyr-sodium, naptalam and naptalam-sodium; b15) from the group of the other herbicides: bromobutide, chlorflurenol, chlorflurenol-methyl, cinmethylin, cumyluron, cyclopyrimorate (CAS 499223-49-3) and its salts and esters, dalapon, dazomet, difenzoquat, difenzoquat-metilsulfate, dimethipin, DSMA, dymron, endothal and its salts, etobenzanid, flamprop, flamprop-isopropyl, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl, flurenol, flurenol-butyl, flurprimidol, fosamine, fosamine-ammonium, indanofan, indaziflam, maleic hydrazide, mefluidide, metam, methiozolin (CAS 403640-27-7), methyl azide, methyl bromide, methyl-dymron, methyl iodide, MSMA, oleic acid, oxaziclomefone, pelargonic acid, pyributicarb, quinoclamine, triaziflam and tridiphane..

Most preferred herbicides B that can be used in combination with herbicides A are given in 3 below:

Table 3

Herbicide B

B.1 clethodim

B.2 clodinafop-propargyl

B.3 cycloxydim

B.4 pinoxaden

B.5 sethodydim

B.6 paraquat

B.7 carfentrazone-ethyl

B.8 flumioxazin

B.9 lactofen

B.10 oxadiargyl

B.1 1 oxyflurofen

B.12 saflufenacil

B.13 sulfentrazone

B.14 1 ,5-dimethyl-6-thioxo-3-(2,2,7- trifluoro-3-oxo-4-(prop-2-ynyl)-

3,4-dihydro-2H- benzo[b][1 ,4]oxazin-6-yl)-1 ,3,5- triazinane-2,4-dione

B.15 benzobicylone

B.16 bicyclopyrone

B.17 clomazone

B.18 difufenican

B.19 isoxaflutole

B.20 mesotrione

B.21 norflurazon

B.22 picolinafen

B.23 sulcotrione

B.24 tembotrione

B.25 topramezone

B.26 glyphosate

B.27 glufosinate

B.28 indaziflam

B.29 isoxaben

B.30 dinoseb Any of the herbicides B.1 to B.39 having a carboxylic, amino and/or hydroxyl group are to be understood as to be employed as such and/or in form of agriculturally acceptable salts thereof and/or in form of agriculturally acceptable esters or amides thereof. According to a preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide B.

According to another preferred embodiment of the invention, the composition comprises at least two, preferably exactly two herbicides B different from each other.

According to another preferred embodiment of the invention, the composition comprises at least three, preferably exactly three herbicides B different from each other.

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A and at least one, preferably exactly one, herbicide B.

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A and at least two, preferably exactly two, herbicides B different from each other.

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A and at least three, preferably exactly three, herbicides B different from each other. According to one preferred embodiment of the invention, the composition comprises at least one herbicide B selected from compounds of groups b1 ), b3), b4), b5), b6), b7), b8), b1 1 ), b12), b13), b14) and b15), preferably selected from compounds of groups b1 ), b3), b4), b5), b6), b7), b8), b1 1 ), b12), b13), b14) or b15). According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A, preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b1 ), in particular selected from clethodim (B.1 ), clodinafop-propargyl (B.2), cycloxydim (B.3), pinoxaden (B.4) and sethodydim (B.5).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b3), in particular paraquat and agriculturally acceptable salts thereof (B.6).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b4), in particular selected from carfentrazone-ethyl (B.7), flumioxazin (B.8), lactofen (B.9), oxadiargyl (B.10), oxyflurofen (B.1 1 ), saflufenacil (B.12), sulfentrazone (B.13) and 1 ,5-dimethyl- 6-thioxo-3-(2,2,7-trifluoro-3-oxo-4-(prop-2-ynyl)-3,4-dihydro-2H-benzo[b][1 ,4]oxazin-6-yl)-1 ,3,5- triazinane-2,4-dione (CAS 1258836-72-4) (B.14).

According to another preferred embodiment of the invention, the composition comprises at leats one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b5), preferably selected from benzobicyclone (B.15), bicyclopyrone (B.16), clomazone (B.17), diflufenican (B.18), isoxaflutole (B.19), mesotrione (B.20), norflurazon (B.21 ), picolinafen (B.22), sulcotrione (B.23), tembotrione (B.24), and topramezone (B.25), in particular selected from benzobicyclone (B.15), isoxaflutole (B.19), mesotrione (B.20) and topramezone (B.25).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b6), in particular glyphosate and agriculturally acceptable salts thereof (B.26).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b7), in particular glufosinate and agriculturally acceptable salts thereof (B.27). According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), at least one, preferably exactly one herbicide B selected from the compounds of group b1 1 ), in particular selected from indaziflam (B.28) and isoxaben (B.29).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), at least one, preferably exactly one herbicide B selected from the compounds of group b12), in particular dinoseb (B.30).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b13), in particular from 2,4D and agriculturally acceptable salts thereof (B.31 ), dicamba and agriculturally acceptable salts thereof (B.32), fluroxypyr (B.33), MCPA and agriculturally acceptable salts thereof (B.34), quinmerac and agriculturally acceptable salts thereof (B.35) and quinclorac and agriculturally acceptable salts thereof (B.36).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b14), in particular diflufenzopyr and agriculturally acceptable salts thereof (B.37).

According to another preferred embodiment of the invention, the composition comprises at least one, preferably exactly one herbicide A selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) and a1 1 ), most preferably selected from a1 ), a2), a3), a4), a5), a6), a7), a8), a9), a10) or a1 1 ), and at least one, preferably exactly one herbicide B selected from the compounds of group b15), in particular selected from dymron (= daimuron) (B.38) and pelargonic acid and agriculturally acceptable salts thereof (B.39).

In another embodiment, compositions according to the invention comprise at least two herbicides B, whereby a first herbicide B is selected from the compounds of groups b1 ), b3), b4), b5), b8), b1 1 ), b12, b13), b14 and b15), preferably selected from the compounds of groups b1 ), b3), b4), b5), b8), b1 1 ), b12, b13), b14 or b15) and a second herbicide B is selected from the compounds of groups b6) and b7), preferably selected from the compounds of groups b6) or b7).

Moreover, it may be useful to apply the TK-inhibiting herbicides, when used in combination with a compound B described SUPRA, in combination with safeners. Safeners are chemical compounds which prevent or reduce damage on useful plants without having a major impact on the herbicidal action of herbicides towards unwanted plants. They can be applied either before sowings (e.g. on seed treatments, shoots or seedlings) or in the pre- emergence application or post-emergence application of the useful plant. Furthermore, the safeners C, the TK-inhibiting herbicides and/or the herbicides B can be applied simultaneously or in succession.

Suitable safeners are e.g. (quinolin-8-oxy)acetic acids, 1 -phenyl-5-haloalkyl-1 H-1 ,2,4- triazol-3-carboxylic acids, 1 -phenyl-4,5-dihydro-5-alkyl-1 H-pyrazol-3,5-dicarboxylic acids, 4,5-dihydro-5,5-diaryl-3-isoxazol carboxylic acids, dichloroacetamides, alpha- oximinophenylacetonitriles, acetophenonoximes, 4,6-dihalo-2-phenylpyrimidines, N-[[4- (aminocarbonyl)phenyl]sulfonyl]-2-benzoic amides, 1 ,8-naphthalic anhydride, 2-halo-4- (haloalkyl)-5-thiazol carboxylic acids, phosphorthiolates and N-alkyl-O-phenylcarbamates and their agriculturally acceptable salts and their agriculturally acceptable derivatives such amides, esters, and thioesters, provided they have an acid group.

Examples of preferred safeners C are benoxacor, cloquintocet, cyometrinil, cyprosulfamide, dichlormid, dicyclonon, dietholate, fenchlorazole, fenclorim, flurazole, fluxofenim, furilazole, isoxadifen, mefenpyr, mephenate, naphthalic anhydride, oxabetrinil, 4-(dichloroacetyl)-1 - oxa-4-azaspiro[4.5]decane (MON4660, CAS 71526-07-3) and 2,2,5-trimethyl-3- (dichloroacetyl)-1 ,3-oxazolidine (R-29148, CAS 52836-31 -4).

Especially preferred safeners C are benoxacor, cloquintocet, cyprosulfamide, dichlormid, fenchlorazole, fenclorim, flurazole, fluxofenim, furilazole, isoxadifen, mefenpyr, naphthalic anhydride, oxabetrinil, 4-(dichloroacetyl)-1 -oxa-4-azaspiro[4.5]decane (MON4660, CAS 71526-07-3) and 2,2,5-trimethyl-3-(dichloroacetyl)-1 ,3-oxazolidine (R-29148, CAS 52836- 31 -4).

Particularly preferred safeners C are benoxacor, cloquintocet, cyprosulfamide, dichlormid, fenchlorazole, fenclorim, furilazole, isoxadifen, mefenpyr, naphtalic anhydride, 4- (dichloroacetyl)-1 -oxa-4-azaspiro[4.5]decane (MON4660, CAS 71526-07-3), and 2,2,5- trimethyl-3-(dichloroacetyl)-1 ,3-oxazolidine (R-29148, CAS 52836-31 -4).

Also preferred safeners C are benoxacor, cloquintocet, cyprosulfamide, dichlormid, fenchlorazole, fenclorim, furilazole, isoxadifen, mefenpyr, 4-(dichloroacetyl)-1 -oxa-4- azaspiro[4.5]decane (MON4660, CAS 71526-07-3) and 2,2,5-trimethyl-3-(dichloroacetyl)- 1 ,3-oxazolidine (R-29148, CAS 52836-31 -4)..

Particularly preferred safeners C, which, as component C, are constituent of the

composition according to the invention are the safeners C as defined above; in particular the safeners C.1 - C.12 listed below in table C: Table C

Safener C

C.1 benoxacor

C.2 cloquintocet

C.3 cyprosulfamide

C.4 dichlormid

C.5 fenchlorazole

C.6 fenclorim

C.1 furilazole

C.8 isoxadifen

C.9 mefenpyr

C.10 naphtalic acid anhydride

C.1 1 4-(dichloroacetyl)-1 -oxa-4-azaspiro[4.5]decane (MON4660, CAS 71526-07-3) 2,2,5-trimethyl-3-(dichloro-acetyl)-1 ,3-oxazolidine (R-29148, CAS 52836-31-4)

The TK-inhibiting herbicides (compounds A) and the active compounds B of groups b1 ) to b15) and the active compounds C are known herbicides and safeners, see, for example, The Compendium of Pesticide Common Names (http://www.alanwood.net/pesticides/); Farm Chemicals Handbook 2000 volume 86, Meister Publishing Company, 2000; B. Hock, C. Fedtke, R. R. Schmidt, Herbizide [Herbicides], Georg Thieme Verlag, Stuttgart 1995; W. H. Ahrens, Herbicide Handbook, 7th edition, Weed Science Society of America, 1994; and K. K. Hatzios, Herbicide Handbook, Supplement for the 7th edition, Weed Science Society of America, 1998. 2,2,5-Trimethyl-3-(dichloroacetyl)-1 ,3-oxazolidine [CAS No. 52836-31 -4] is also referred to as R-29148. 4-(Dichloroacetyl)-1 -oxa-4-azaspiro[4.5]decane [CAS No. 71526-07-3] is also referred to as AD-67 and MON 4660.

The assignment of the active compounds to the respective mechanisms of action is based on current knowledge. If several mechanisms of action apply to one active compound, this substance was only assigned to one mechanism of action.

Active compounds B and C having a carboxyl group can be employed in the form of the acid, in the form of an agriculturally suitable salt as mentioned above or else in the form of an agriculturally acceptable derivative in the compositions according to the invention.

In the case of dicamba, suitable salts include those, where the counterion is an agriculturally acceptable cation. For example, suitable salts of dicamba are dicamba-sodium, dicamba-potassium, dicamba-methylammonium, dicamba-dimethylammonium, dicamba- isopropylammonium, dicamba-diglycolamine, dicamba-olamine, dicamba-diolamine, dicamba-trolamine, dicamba-N,N-bis-(3-aminopropyl)methylamine and dicamba- diethylenetriamine. Examples of a suitable ester are dicamba-methyl and dicamba-butotyl. Suitable salts of 2,4-D are 2,4-D-ammonium, 2,4-D-dimethylammonium, 2,4-D- diethylammonium, 2,4-D-diethanolammonium (2,4-D-diolamine), 2,4-D-triethanol- ammonium, 2,4-D-isopropylammonium, 2,4-D-triisopropanolammonium, 2,4-D- heptylammonium, 2,4-D-dodecylammonium, 2,4-D-tetradecylammonium, 2,4-D- triethylammonium, 2,4-D-tris(2-hydroxypropyl)ammonium, 2,4-D-tris(isopropyl)ammonium, 2,4-D-trolamine, 2,4-D-lithium, 2,4-D-sodium. Examples of suitable esters of 2,4-D are 2,4- D-butotyl, 2,4-D-2-butoxypropyl, 2,4-D-3-butoxypropyl, 2,4-D-butyl, 2,4-D-ethyl, 2,4-D- ethylhexyl, 2,4-D-isobutyl, 2,4-D-isooctyl, 2,4-D-isopropyl, 2,4-D-meptyl, 2,4-D-methyl, 2,4- D-octyl, 2,4-D-pentyl, 2,4-D-propyl, 2,4-D-tefuryl and clacyfos.

Suitable salts of 2,4-DB are for example 2,4-DB-sodium, 2,4-DB-potassium and 2,4-DB- dimethylammonium. Suitable esters of 2,4-DB are for example 2,4-DB-butyl and 2,4-DB- isoctyl.

Suitable salts of dichlorprop are for example dichlorprop-sodium, dichlorprop-potassium and dichlorprop-dimethylammonium. Examples of suitable esters of dichlorprop are dichlorprop- butotyl and dichlorprop-isoctyl. Suitable salts and esters of MCPA include MCPA-butotyl, MCPA-butyl, MCPA-dimethyl- ammonium, MCPA-diolamine, MCPA-ethyl, MCPA-thioethyl, MCPA-2-ethylhexyl, MCPA- isobutyl, MCPA-isoctyl, MCPA-isopropyl, MCPA-isopropylammonium, MCPA-methyl, MCPA-olamine, MCPA-potassium, MCPA-sodium and MCPA-trolamine.

A suitable salt of MCPB is MCPB sodium. A suitable ester of MCPB is MCPB-ethyl.

Suitable salts of clopyralid are clopyralid-potassium, clopyralid-olamine and clopyralid-tris- (2-hydroxypropyl)ammonium. Example of suitable esters of clopyralid is clopyralid-methyl. Examples of a suitable ester of fluroxypyr are fluroxypyr-meptyl and fluroxypyr-2-butoxy-1 - methylethyl, wherein fluroxypyr-meptyl is preferred.

Suitable salts of picloram are picloram-dimethylammonium, picloram-potassium, picloram- triisopropanolammonium, picloram-triisopropylammonium and picloram-trolamine. A suitable ester of picloram is picloram-isoctyl.

A suitable salt of triclopyr is triclopyr-triethylammonium. Suitable esters of triclopyr are for example triclopyr-ethyl and triclopyr-butotyl.

Suitable salts and esters of chloramben include chloramben-ammonium, chloramben- diolamine, chloramben-methyl, chloramben-methylammonium and chloramben-sodium. Suitable salts and esters of 2,3,6-TBA include 2,3,6-TBA-dimethylammonium, 2,3,6-TBA- lithium, 2,3,6-TBA-potassium and 2,3,6-TBA-sodium.

Suitable salts and esters of aminopyralid include aminopyralid-potassium and aminopyralid- tris(2-hydroxypropyl)ammonium.

Suitable salts of glyphosate are for example glyphosate-ammonium, glyphosate- diammonium, glyphoste-dimethylammonium, glyphosate-isopropylammonium, glyphosate- potassium, glyphosate-sodium, glyphosate-trimesium as well as the ethanolamine and diethanolamine salts, preferably glyphosate-diammonium, glyphosate-isopropylammonium and glyphosate-trimesium (sulfosate).

A suitable salt of glufosinate is for example glufosinate-ammonium.

A suitable salt of glufosinate-P is for example glufosinate-P-ammonium.

Suitable salts and esters of bromoxynil are for example bromoxynil-butyrate, bromoxynil- heptanoate, bromoxynil-octanoate, bromoxynil-potassium and bromoxynil-sodium.

Suitable salts and esters of ioxonil are for example ioxonil-octanoate, ioxonil-potassium and ioxonil-sodium.

Suitable salts and esters of mecoprop include mecoprop-butotyl, mecoprop- dimethylammonium, mecoprop-diolamine, mecoprop-ethadyl, mecoprop-2-ethylhexyl, mecoprop-isoctyl, mecoprop-methyl, mecoprop-potassium, mecoprop-sodium and mecoprop-trolamine.

Suitable salts of mecoprop-P are for example mecoprop-P-butotyl, mecoprop-P- dimethylammonium, mecoprop-P-2-ethylhexyl, mecoprop-P-isobutyl, mecoprop-P- potassium and mecoprop-P-sodium.

A suitable salt of diflufenzopyr is for example diflufenzopyr-sodium.

A suitable salt of naptalam is for example naptalam-sodium.

Suitable salts and esters of aminocyclopyrachlor are for example aminocyclopyrachlor- dimethylammonium, aminocyclopyrachlor-methyl, aminocyclopyrachlor- triisopropanolammonium, aminocyclopyrachlor-sodium and aminocyclopyrachlor- potassium.

A suitable salt of quinclorac is for example quinclorac-dimethylammonium.

A suitable salt of quinmerac is for example quinclorac-dimethylammonium.

A suitable salt of imazamox is for example imazamox-ammonium.

Suitable salts of imazapic are for example imazapic-ammonium and imazapic- isopropylammonium.

Suitable salts of imazapyr are for example imazapyr-ammonium and imazapyr- isopropylammonium.

A suitable salt of imazaquin is for example imazaquin-ammonium.

Suitable salts of imazethapyr are for example imazethapyr-ammonium and imazethapyr- isopropylammonium.

A suitable salt of topramezone is for example topramezone-sodium. The preferred embodiments of the invention mentioned herein below have to be understood as being preferred either independently from each other or in combination with one another.

According to a preferred embodiment of the invention, the composition comprises as component B at least one, preferably exactly one herbicide B.

According to another preferred embodiment of the invention, the composition comprises at least two, preferably exactly two, herbicides B different from each other.

According to another preferred embodiment of the invention, the composition comprises at least three, preferably exactly three, herbicides B different from each other.

Here and below, the term "binary compositions" includes compositions comprising one or more, for example 1 , 2 or 3, herbicides A and either one or more, for example 1 , 2 or 3, herbicides B. Correspondingly, the term "ternary compositions" includes compositions comprising one or more, for example 1 , 2 or 3, herbicides A, for example 1 , 2 or 3, herbicides B and at least one herbicide D.

Further preferred embodiments relate to compositions which correspond to the binary and ternary compositions mentioned above and additionally comprise a safener C, in particular selected from the group consisting of benoxacor (C.1 ), cloquintocet (C.2), cyprosulfamide (C.3), dichlormid (C.4), fenchlorazole (C.5), fenclorim (C.6), furilazole (C.7), isoxadifen (C.8), mefenpyr (C.9), 4-(dichloroacetyl)-1 -oxa-4-azaspiro[4.5]decane (MON4660, CAS 71526-07-3) (C.10) and 2,2,5-trimethyl-3-(dichloroacetyl)-1 ,3-oxazolidine (R-29148, CAS 52836-31 -4) (C.1 1 ). In binary compositions comprising at least one herbicide A and at least one herbicide B, the weight ratio of A:B is generally in the range of from 1 :1000 to 1000:1 , preferably in the range of from 1 :500 to 500:1 , in particular in the range of from 1 :250 to 250:1 and particularly preferably in the range of from 1 :75 to 75: 1.

In compositions comprising both at least one herbicide A, at least one herbicide B and at least one safener C, the weight ratio A:B is generally in the range of from 1 :1000 to 1000:1 , preferably in the range of from 1 :500 to 500:1 , in particular in the range of from 1 :250 to 250:1 and particularly preferably in the range of from 1 :75 to 75:1 ; the weight ratio A:C is generally in the range of from 1 :1000 to 1000:1 , preferably in the range of from 1 :500 to 500:1 , in particular in the range of from 1 :250 to 250:1 and particularly preferably in the range of from 1 :75 to 75:1 ; and the weight ratio B:C is generally in the range of from 1 :1000 to 1000:1 , preferably in the range of from 1 :500 to 500:1 , in particular in the range of from 1 :250 to 250:1 and particularly preferably in the range of from 1 :75 to 75:1. The weight ratio (A + B):C is preferably in the range of from 1 :500 to 500:1 , in particular in the range of from 1 :250 to 250:1 and particularly preferably in the range of from 1 :75 to 75:1 . In ternary compositions comprising at least one herbicide A, at least one herbicide B, and a herbicide D, the weight ratios A:B, A:D and B:D in each case are generally in the range of from 1 :1000 to 1000:1 , preferably in the range of from 1 :500 to 500:1 , in particular in the range of from 1 :250 to 250:1 and particularly preferably in the range of from 1 :75 to 75:1. The weight ratio (A + B):D is preferably in the range of from 1 :500 to 500:1 , in particular in the range of from 1 :250 to 250:1 and particularly preferably in the range of from 1 :75 to 75:1.

It is generally preferred to use the compounds of the invention in combination with herbicides that are selective for the crop being treated and which complement the spectrum of weeds controlled by these compounds at the application rate employed. It is further generally preferred to apply the compounds of the invention and other complementary herbicides at the same time, either as a combination formulation or as a tank mix.

In another embodiment, the present invention refers to a method for identifying a TK-inhibiting herbicide by using a mutated TK encoded by a nucleic acid which comprises the nucleotide sequence of SEQ ID NO: 182 or 183, or a variant or derivative thereof.

Said method comprises the steps of:

a) generating a transgenic cell or plant comprising a nucleic acid encoding a mutated TK, wherein the mutated TK is expressed;

c) determining the growth or the viability of the transgenic cell or plant and the control cell or plant after application of said TK-inhibiting herbicide, and

d) selecting "TK-inhibiting herbicides" which confer reduced growth to the control cell or plant as compared to the growth of the transgenic cell or plant.

As described above, the present invention teaches compositions and methods for increasing the TK-inhibiting tolerance of a crop plant or seed as compared to a wild-type variety of the plant or seed. In a preferred embodiment, the TK-inhibiting tolerance of a crop plant or seed is increased such that the plant or seed can withstand a TK-inhibiting herbicide application of preferably approximately 1-1000 g ai ha^-1, more preferably 1-200 g ai ha^-1, even more preferably 5-150 g ai ha^-1, and most preferably 10-100 g ai ha^-1. As used herein, to "withstand" a TK- inhibiting herbicide application means that the plant is either not killed or only moderately injured by such application. It will be understood by the person skilled in the art that the application rates may vary, depending on the environmental conditions such as temperature or humidity, and depending on the chosen kind of herbicide (active ingredient ai). Post-emergent weed control methods useful in various embodiments hereof utilize about >0.3x application rates of TK-inhibiting herbicides; in some embodiments, this can be about, for example, >0.3x, >0.4x, >0.5x, >0.6x, >0.7x, >0.8x, >0.9x, or >lx of TK-inhibiting herbicides. In one embodiment, TK-inhibiting herbicides-tolerant plants of the present invention have tolerance to a post-emergant application of a TK-inhibiting herbicides at an amount of about 25 to about 200 g ai/ha. In some embodiments, wherein the TK-inhibiting herbicides-tolerant plant is a dicot (e.g. , soy, cotton), the post-emergant application of the TK-inhibiting herbicides is at an amount of about 50 g ai/ha. In another embodiment, wherein the TK-inhibiting herbicides-tolerant plant is a monocot (e.g., maize, rice, sorghum), the post-emergant application of the TK-inhibiting herbicides is at an amount of about 200 g ai/ha. In other embodiments, wherein the TK-inhibiting herbicides-tolerant plant is a

Brassica (e.g., canola), the post-emergant application of the TK-inhibiting herbicides is at an amount of about 25 g ai/ha. In post-emergent weed control methods hereof, in some embodiments, the method can utilize TK-inhibiting herbicides application rates at about 7 to 10 days post-emergent. In another embodiment, the application rate can exceed Ix TK- inhibiting herbicides; in some embodiments, the rate can be up to 4x TK-inhibiting herbicides, though more typically it will be about 2.5x or less, or about 2x or less, or about 1x or less.

Furthermore, the present invention provides methods that involve the use of at least one TK-inhibiting herbicide, optionally in combination with one or more herbicidal compounds B, and, optionally, a safener C, as described in detail supra.

In these methods, the TK-inhibiting herbicide can be applied by any method known in the art including, but not limited to, seed treatment, soil treatment, and foliar treatment. Prior to application, the TK-inhibiting herbicide can be converted into the customary formulations, for example solutions, emulsions, suspensions, dusts, powders, pastes and granules. The use form depends on the particular intended purpose; in each case, it should ensure a fine and even distribution of the compound according to the invention. By providing plants having increased tolerance to TK-inhibiting herbicide, a wide variety of formulations can be employed for protecting plants from weeds, so as to enhance plant growth and reduce competition for nutrients. A TK-inhibiting herbicide can be used by itself for pre-emergence, post-emergence, pre-planting, and at-planting control of weeds in areas surrounding the crop plants described herein, or a TK-inhibiting herbicide formulation can be used that contains other additives. The TK-inhibiting herbicide can also be used as a seed treatment. Additives found in a TK-inhibiting herbicide formulation include other herbicides, detergents, adjuvants, spreading agents, sticking agents, stabilizing agents, or the like. The TK-inhibiting herbicide formulation can be a wet or dry preparation and can include, but is not limited to, flowable powders, emulsifiable concentrates, and liquid concentrates. The TK-inhibiting herbicide and herbicide formulations can be applied in accordance with conventional methods, for example, by spraying, irrigation, dusting, or the like.

Suitable formulations are described in detail in PCT/EP2009/063387 and

PCT/EP2009/063386, which are incorporated herein by reference. As disclosed herein, the TK nucleic acids of the invention find use in enhancing the herbicide tolerance of plants that comprise in their genomes a gene encoding a herbicide-tolerant wild- type or mutated TK protein. Such a gene may be an endogenous gene or a transgene, as described above. Additionally, in certain embodiments, the nucleic acids of the present invention can be stacked with any combination of polynucleotide sequences of interest in order to create plants with a desired phenotype. For example, the nucleic acids of the present invention may be stacked with any other polynucleotides encoding polypeptides having pesticidal and/or insecticidal activity, such as, for example, the Bacillus thuringiensis toxin proteins (described in U.S. Patent Nos. 5,366,892; 5,747,450; 5,737,514; 5,723,756; 5,593,881 ; and Geiser et al (1986) Gene 48: 109), 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), Glyphosate acetyl transferase (GAT), cytochrome P450 monooxygenase, phosphinothricin acetyltransferase (PAT), Acetohydroxyacid synthase (AHAS; EC 4.1 .3.18, also known as acetolactate synthase or ALS), hydroxyphenyl pyruvate dioxygenase (HPPD), Phytoene desaturase (PD), Protoporphyrinogen oxidase (PPO) and dicamba degrading enzymes as disclosed in WO 02/068607, or phenoxyaceticacid- and phenoxypropionicacid-derivative degrading enzymes as disclosed in WO 2008141 154 or WO 2005107437. The combinations generated can also include multiple copies of any one of the polynucleotides of interest.

Consequently, Herbicide-tolerant plants of the invention can be used in conjunction with an herbicide to which they are tolerant. Herbicides can be applied to the plants of the invention using any techniques known to those skilled in the art. Herbicides can be applied at any point in the plant cultivation process. For example, herbicides can be applied pre-planting, at planting, pre-emergence, post-emergence or combinations thereof. Herbicides may be applied to seeds and dried to form a layer on the seeds. In some embodiments, seeds are treated with a safener, followed by a post- emergent application of a TK-inhibiting herbicides. In one embodiment, the post-emergent application of the TK-inhibiting herbicides is about 7 to 10 days following planting of safener-treated seeds. In some embodiments, the safener is cloquintocet, dichlormid, fluxofenim, or combinations thereof.

Methods of controlling weeds or undesired vegetation

In other aspects, the present invention provides a method for controlling weeds at a locus for growth of a plant or plant part thereof, the method comprising: applying a composition comprising a TK-inhibiting herbicides to the locus. In some aspects, the present invention provides a method for controlling weeds at a locus for growth of a plant, the method comprising: applying an herbicide composition comprising TK-inhibiting herbicides to the locus; wherein said locus is: (a) a locus that contains: a plant or a seed capable of producing said plant; or (b) a locus that is to be after said applying is made to contain the plant or the seed; wherein the plant or the seed comprises in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides. Herbicide compositions hereof can be applied, e.g., as foliar treatments, soil treatments, seed treatments, or soil drenches. Application can be made, e.g., by spraying, dusting, broadcasting, or any other mode known useful in the art.

In one embodiment, herbicides can be used to control the growth of weeds that may be found growing in the vicinity of the herbicide-tolerant plants invention. In embodiments of this type, an herbicide can be applied to a plot in which herbicide-tolerant plants of the invention are growing in vicinity to weeds. An herbicide to which the herbicide-tolerant plant of the invention is tolerant can then be applied to the plot at a concentration sufficient to kill or inhibit the growth of the weed. Concentrations of herbicide sufficient to kill or inhibit the growth of weeds are known in the art and are disclosed above.

In other embodiments, the present invention provides a method for controlling weeds in the vicinity of a TK-inhibiting herbicides-tolerant plant of the invention. The method comprises applying an effective amount of a TK-inhibiting herbicides to the weeds and to the auxinic herbicide- tolerant plant, wherein the plant has increased tolerance to auxinic herbicide when compared to a wild-type plant. In some embodiments, the TK-inhibiting herbicides- tolerant plants of the invention are preferably crop plants, including, but not limited to, sunflower, alfalfa, Brassica sp., soybean, cotton, safflower, peanut, tobacco, tomato, potato, wheat, rice, maize, sorghum, barley, rye, millet, and sorghum.

In other aspects, herbicide(s) (e.g., TK-inhibiting herbicides) can also be used as a seed treatment. In some embodiments, an effective concentration or an effective amount of herbicide(s), or a composition comprising an effective concentration or an effective amount of herbicide(s) can be applied directly to the seeds prior to or during the sowing of the seeds. Seed Treatment formulations may additionally comprise binders and optionally colorants.

Binders can be added to improve the adhesion of the active materials on the seeds after treatment. In one embodiments, suitable binders are block copolymers EO/PO surfactants but also polyvinylalcoholsl, polyvinylpyrrolidones, polyacrylates, polymethacrylates, polybutenes, polyisobutylenes, polystyrene, polyethyleneamines, polyethyleneamides, polyethyleneimines (Lupasol(R), Polymin(R)), polyethers, polyurethans, polyvinylacetate, tylose and copolymers derived from these polymers. Optionally, also colorants can be included in the formulation. Suitable colorants or dyes for seed treatment formulations are Rhodamin B, C.I. Pigment Red 1 12, C.I. Solvent Red 1 , pigment blue 15:4, pigment blue 15:3, pigment blue 15:2, pigment blue 15: 1 , pigment blue 80, pigment yellow 1 , pigment yellow 13, pigment red 1 12, pigment red 48:2, pigment red 48: 1 , pigment red 57: 1 , pigment red 53:1 , pigment orange 43, pigment orange 34, pigment orange 5, pigment green 36, pigment green 7, pigment white 6, pigment brown 25, basic violet 10, basic violet 49, acid red 51 , acid red 52, acid red 14, acid blue 9, acid yellow 23, basic red 10, basic red 108.

The term seed treatment comprises all suitable seed treatment techniques known in the art, such as seed dressing, seed coating, seed dusting, seed soaking, and seed pelleting. In one embodiment, the present invention provides a method of treating soil by the application, in particular into the seed drill: either of a granular formulation containing the TK-inhibiting herbicides as a composition/formulation (e.g., a granular formulation), with optionally one or more solid or liquid, agriculturally acceptable carriers and/or optionally with one or more agriculturally acceptable surfactants. This method is advantageously employed, for example, in seedbeds of cereals, maize, cotton, and sunflower. The present invention also comprises seeds coated with or containing with a seed treatment formulation comprising TK-inhibiting herbicides and at least one other herbicide such as, e.g. , an AHAS-inhibitor selected from the group consisting of amidosulfuron, azimsulfuron, bensulfuron, chlorimuron, chlorsulfuron, cinosulfuron, cyclosulfamuron, ethametsulfuron, ethoxysulfuron, flazasulfuron, flupyrsulfuron, foramsulfuron, halosulfuron, imazosulfuron, iodosulfuron, mesosulfuron, metsulfuron, nicosulfuron, oxasulfuron, primisulfuron, prosulfuron, pyrazosulfuron, rimsulfuron, sulfometuron, sulfosulfuron, thifensulfuron, triasulfuron, tribenuron, trifloxysulfuron, triflusulfuron, tritosulfuron,

imazamethabenz, imazamox, imazapic, imazapyr, imazaquin, imazethapyr, cloransulam, diclosulam, florasulam, flumetsulam, metosulam, penoxsulam, bispyribac, pyriminobac, propoxycarbazone, flucarbazone, pyribenzoxim, pyriftalid and pyrithiobac.

The term "coated with and/or containing" generally signifies that the active ingredient is for the most part on the surface of the propagation product at the time of application, although a greater or lesser part of the ingredient may penetrate into the propagation product, depending on the method of application. When the said propagation product is (re)planted, it may absorb the active ingredient.

In some embodiments, the seed treatment application with TK-inhibiting herbicides or with a formulation comprising the TK-inhibiting herbicides is carried out by spraying or dusting the seeds before sowing of the plants and before emergence of the plants. In other embodiments, in the treatment of seeds, the corresponding formulations are applied by treating the seeds with an effective amount of TK-inhibiting herbicides or a formulation comprising the TK-inhibiting herbicides.

In other aspects, the present invention provides a method for combating undesired vegetation or controlling weeds comprising contacting the seeds of the TK-inhibiting herbicides-tolerant plants of the present invention before sowing and/or after

pregermination with TK-inhibiting herbicides. The method can further comprise sowing the seeds, for example, in soil in a field or in a potting medium in greenhouse. The method finds particular use in combating undesired vegetation or controlling weeds in the immediate vicinity of the seed. The control of undesired vegetation is understood as the killing of weeds and/or otherwise retarding or inhibiting the normal growth of the weeds. Weeds, in the broadest sense, are understood as meaning all those plants which grow in locations where they are undesired. The weeds of the present invention include, for example, dicotyledonous and

monocotyledonous weeds. Dicotyledonous weeds include, but are not limited to, weeds of the genera: Sinapis, Lepiclium, Galium, Stellaria, Matricaria, Anthemis, Galinsoga,

Chenopodium, Urtica, Senecio, Amaranthus, Portulaca, Xanthium, Convolvulus, Ipomoea, Polygonum, Sesbania, Ambrosia, Cirsium, Carduus, Sonchus, Solarium, Rorippa, Rotala, Lindernia, Lamium, Veronica, Abutilon, Emex, Datura, Viola, Galeopsis, Papaver,

Centaurea, Trifolium, Ranunculus, and Taraxacum. Monocotyledonous weeds include, but are not limited to, weeds of the genera: Echinochloa, Setaria, Panicum, Digitaria, Phleum, Poa, Festuca, Eleusine, Brachiaria, Lolium, Bromus, Avena, Cyperus, Sorghum, Agropyron, Cynodon, Monochoria, Fimbristyslis, Sagittaria, Eleocharis, Scirpus, Paspalum,

Ischaemum, Sphenoclea, Dactyloctenium, Agrostis, Alopecurus, and Apera.

In addition, the weeds of the present invention can include, for example, crop plants that are growing in an undesired location. For example, a volunteer maize plant that is in a field that predominantly comprises soybean plants can be considered a weed, if the maize plant is undesired in the field of soybean plants.

In other embodiments, in the treatment of seeds, the corresponding formulations are applied by treating the seeds with an effective amount of TK-inhibiting herbicides or a formulation comprising the TK-inhibiting herbicides.

In still further aspects, treatment of loci, plants, plant parts, or seeds of the present invention comprises application of an agronomically acceptable composition that does not contain an A.I. In one embodiment, the treatment comprises application of an agronomically

acceptable composition that does not contain a TK-inhibiting herbicides A.I. In some embodiments, the treatment comprises application of an agronomically acceptable composition that does not contain a TK-inhibiting herbicides A.L, wherein the composition comprises one or more of agronomically-acceptable carriers, diluents, excipients, plant growth regulators, and the like. In other embodiments, the treatment comprises application of an agronomically acceptable composition that does not contain a TK-inhibiting herbicides A.I., wherein the composition comprises an adjuvant. In one embodiment, the adjuvant is a surfactant, a spreader, a sticker, a penetrant, a drift-control agent, a crop oil, an emulsifier, a compatibility agent, or combinations thereof.

It should also be understood that the foregoing relates to preferred embodiments of the present invention and that numerous changes may be made therein without departing from the scope of the invention. The invention is further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope thereof. On the contrary, it is to be clearly understood that resort may be had to various other embodiments, modifications, and equivalents thereof, which, after reading the description herein, may suggest themselves to those skilled in the art without departing from the spirit of the present invention and/or the scope of the appended claims. EXAMPLES

EXAMPLE 1 : Sequencing and full length assembly of transketolase genes

Isolation of RNA and cDNA synthesis

Plant leaf tissue is harvested, frozen and grounded in liquid nitrogen and total RNA is extracted using an Ambion RNAqueous-Midi kit (AM191 1 , Ambion) with the Plant RNA Isolation Aid (AM9690, Ambion) as per manufacturer's recommendation. The last elution is done with 10 ul of elution solution. To validate the quality of the extracted RNA 1 uL of the final product is run on a Bioanalyzer 2100 using the RNA 6000 Nano kit with the Plant RNA Nano method. The final solution, containing purified RNA, is stored at -80° C until library preparation.

For assembly of plant genes, an RNA sequencing experiment is performed. RNA sequencing libraries are produced using TruSeq RNA Sample preparation kits V2 (RS-122-2001 ) from lllumina according to the instructions of the manufacturer. Briefly, 1 μg of total RNA is first purified twice on a poly-dT column. During the second elution step, RNA is fragmented and primed for cDNA synthesis. The material is reverse transcribed, RNA is removed and the second strand is produced. After rendering the ends of the fragment blunt, 3' ends are adenylated and lllumina sequencing-specific bar-coded adaptors are ligated at both ends of the fragments. The DNA fragments bearing adaptors at both ends are enriched by a 15 cycle PCR amplification. Libraries are pooled prior to sequencing. The pooled libraries are first put on a flowcell using a TruSeq PE Cluster kit V3 (PE-401-3001 ) on the cBot and clusters are amplified on the device. Afterwards, the flowcell is transferred onto the lllumina Hiseq machine and the material on the flowcell is then sequenced using lllumina TruSeq SBS Kit V3 (FC-401 -3001 ) as per manufacturer's recommendation.

The data produced by the lllumina Hiseq sequencer is first trimmed at both ends using a quality threshold of 15 using the FASTQC Quality Trimmer

(http://www.bioinformatics.babraham.ac.uk/projects/fastqc/). These sequences are further analyzed to remove any lllumina adaptor sequences using CutAdapt

(http://code.google.eom/p/cutadapt/). Sequence reads are assembled using CLC bio algorithm (version 4.01 ). The transketolase full length gene sequences (SEQID NO 182 and 183) are identified by performing a BLAST search with Arabidopsis thaliana transketolase sequences as query and the CLC bio assembly as database. EXAMPLE 2: Mutagenesis of the Gene Encoding for Transketolase

All nucleic acid coding sequence and all single and double mutants based on SEQ ID NO 183 were synthesized and cloned by Geneart (Geneart AG, Regensburg, Germany). Rational design mutants were synthesized by Geneart. Random transketolase gene libraries were synthesized by Geneart. Plasmids were isolated from E. coli TOP10 by performing a plasmid minpreparation and confirmed by DNA sequencing.

EXAMPLE 3: Expression and purification of recombinant wild-type and mutant Transketolase

Clones in pET27b vector were transformed into BL21 (DE3)-pLysS strain of E. coli. Cells were grown in 250 - 1000 mL of LB with 100 μgmL-1 of ampicillin or kanamycin, shaking overnight at 37 °C. The cells were harvested by centrifugation at 1600*g, washed with 0.09% NaCI, and stored at -80 °C. Cells were lysed using a French press at 140 MPa in 50 mM sodium phosphate pH 7.5, 1 M NaCI, 5 mM imidazole, 5% glycerol, and 1 μg mL-1 leupeptin. Following lysis, 0.5 U of Benzonase (Novagen, EMD Chemicals, Inc., Gibbstown, NJ) and PMSF (final concentration of 1 mM) were added. Cell debris was removed by centrifugation at 3000*g. His- tagged transketolase proteins were purified on a nickel activated Hitrap Chelating HP column (GE Healthcare Bio-Sciences Corp., Piscataway, NJ) equilibrated with 20 mM sodium phosphate pH 8.0, 50 mM NaCI, 5 mM imidazole, 5 mM MgCI2, 0.1 mM EDTA, and 17% glycerol. Target ptotein is eluted with 250 mM imidazole. The active protein was desalted on a PD-10 column (GE Healthcare Bio-Sciences Corp., Piscataway, NJ) equilibrated with a 20 mM sodium phosphate buffer, pH 7.5, 5 mM MgCI2, 1 mM EDTA and 17% glycerol.

EXAMPLE 4: Wild-type and mutant Transketolase Enzyme and Inhibitor Assay

Assays were measured for 1 hour at 340 nm using a standard plate reader. Before starting the reactions with Xylulose-5-Phosphat, the reaction mixture was incubated for 15 minutes at room temperature. Both Cornexistin and Hydroxycornexistin were used as inhibitors of transketolase activity. Dose response curves were measured by titrating either of the inhibitors. On the basis of dose response curves, IC50 values were calculated and use to evaluate in vitro tolerance to either inhibitor. (IC50 is defined as the concentration of inhibitor needed to induce 50% loss in enzyme activity as compared to a similar reaction without inhibitor.)

The results are shown in Table 4

Table 4

Tolerance

Substitution Substitution Substitution Relative IC50 (M)

Factor

Position 1 Position 2 Position 3 Activity Comexistin

(wild type = 1,0)

- - - 4,00 4,46E-06 1,0

M463A L544C - 0,41 6,58E-04 147,5

V514N L544V - 4,41 >1,00E-4 >22,4

T511S V514N L544V 5,10 >1,00E-4 >22,4

M463C L544V - 0,71 >1,00E-4 >22,4

M463G L544S - 0,47 >1,00E-4 >22,4

M463G L544T - 0,74 >1,00E-4 >22,4

M463G L544V - 0,49 >1,00E-4 >22,4

M463G L544C - 0,50 >1,00E-4 >22,4

M463G V514N L544T 0,49 >1,00E-4 >22,4

M463G V514N L544C 0,38 >1,00E-4 >22,4

M463G V514N L544S 0,32 >1,00E-4 >22,4

M463G V514N L544V 0,44 >1,00E-4 >22,4

M463G T511S L544T 1,18 >1,00E-4 >22,4

M463G T511S L544C 0,36 >1,00E-4 >22,4

M463G T511S L544S 0,49 >1,00E-4 >22,4

M463C T511S L544T 1,47 >1,00E-4 >22,4

M463C T511S L544C 0,52 >1,00E-4 >22,4

M463C T511S L544S 0,30 >1,00E-4 >22,4

M463C V514N L544T 2,37 >1,00E-4 >22,4

M463C V514N L544C 0,20 >1,00E-4 >22,4

M463C V514N L544S 0,36 >1,00E-4 >22,4

M463C V514N L544V 0,33 >1,00E-4 >22,4

Y337D - - 4,99 1,61E-04 36,2

M463C L544T - 2,89 1 ,57E-04 35,3

M463C L544A - 0,49 1,00E-04 22,4

M463A L544A - 0,29 1,00E-04 22,4

M463A L544C - 0,18 1,00E-04 22,4

M463A L544S - 0,10 1,00E-04 22,4

M463C L544C - 2,01 9,07E-05 20,4

Y337I - - 5,33 8,32E-05 18,7

M463L - - 4,54 8,03E-05 18,0

S460A - - 5,73 7,94E-05 17,8

L544C - - 2,94 7,78E-05 17,5

V514N - - 9,83 7,68E-05 17,2

M463K - - 6,01 7,29E-05 16,4 S461 M - - 0,04 6,16E-05 13,8

Y337N - - 5,58 6,01 E-05 13,5

Y337C - - 7,78 5,99E-05 13,4

M463G - - 9,75 5,94E-05 13,3

S460E - - 5,10 5,82E-05 13,1

F512C - - 0,60 5,76E-05 12,9

Y337L - - 5,1 1 5,60E-05 12,6

L544V - - 8,18 5,30E-05 1 1 ,9

M463N - - 5,09 5,18E-05 1 1 ,6

L458I - - 4,98 4,98E-05 1 1 ,2

L544V - - 5,84 4,98E-05 1 1 ,2

L544I - - 5,31 4,68E-05 10,5

S342D A343P - 0,79 4,61 E-05 10,3

L544G - - 0,55 4,03E-05 9,0

S460L - - 5,58 3,90E-05 8,8

S342D - - 0,84 3,87E-05 8,7

L458V - - 6,41 3,65E-05 8,2

L544H - - 0,09 3,51 E-05 7,9

M463A L544S - 0,1 1 3,46E-05 7,8

V514H - - 0,07 3,44E-05 7,7

M463A L544T - 0,39 3,42E-05 7,7

M463Y - - 0,91 3,18E-05 7,1

F513L - - 5,39 3,03E-05 6,8

S460G - - 7,30 2,98E-05 6,7

M463C L544S - 1 ,55 2,83E-05 6,3

S461A - - 0,47 2,82E-05 6,3

L544Q - 0,57 2,78E-05 6,2

I267C - - 0,36 2,66E-05 6,0

S460Q - - 7,25 2,64E-05 5,9

A343V - - 1 ,28 2,59E-05 5,8

M463F - - 1 ,22 2,59E-05 5,8

Y337F - - 5,52 2,51 E-05 5,6

S460T - - 5,40 2,48E-05 5,6

A343S - - 2,27 2,47E-05 5,5

L458M - - 5,25 2,45E-05 5,5

Y337E - - 7,53 2,40E-05 5,4

L458C - - 1 ,90 2,38E-05 5,3

V514C - - 10,36 2,28E-05 5,1

Y337T - - 5,36 2,26E-05 5,1

M463I - - 0,18 2,26E-05 5,1

T51 1 S - - 6,42 2,24E-05 5,0

Y337A - - 5,51 2,09E-05 4,7 Y337H - - 8,34 1.98E-05 4,4

S460N - - 5,44 1.95E-05 4,4

V514A - - 8,70 1 ,86E-05 4,2

S342T - - 5,25 1 ,86E-05 4,2

S460F - - 5,54 1.80E-05 4,0

I267L - - 1,95 1.79E-05 4,0

A459S - - 7,00 1 ,78E-05 4,0

F513M - - 3,48 1 ,77E-05 4,0

L458S - - 0,04 1.74E-05 3,9

V514M - - 3,85 1.71E-05 3,8

A459C - - 1,80 1.64E-05 3,7

V514T - - 1,54 1.61E-05 3,6

A343T - - 0,81 1 ,58E-05 3,5

F513Y - - 4,64 1 ,55E-05 3,5

Y337V - - 0,87 1 ,55E-05 3,5

L458A - - 0,96 1 ,55E-05 3,5

F513I - - 1,22 1.53E-05 3,4

V514D - - 1,49 1.51E-05 3,4

F513V - - 2,43 1 ,47E-05 3,3

Y337Q - - 10,62 1.42E-05 3,2

S460M - - 5,14 1.39E-05 3,1

A459T - - 5,73 1.39E-05 3,1

S342A - - 5,90 1.38E-05 3,1

V514L - - 0,71 1.38E-05 3,1

Y337M - - 1,17 1.33E-05 3,0

S342N A343P - 3,85 1.32E-05 3,0

Y337S - - 1,09 1.15E-05 2,6

F515A - - 0,11 1.15E-05 2,6

Y337W - - 4,09 1.12E-05 2,5

F512A - - 0,07 1.11E-05 2,5

F515M - - 0,14 1.11E-05 2,5

I267T - - 0,93 1.07E-05 2,4

I267D - - 0,05 1.06E-05 2,4

M463H - - 0,26 1.02E-05 2,3

A459I - - 0,04 1.01E-05 2,3

S342T A343P - 4,97 1.00E-05 2,3

A343C - - 3,70 9,75E-06 2,2

V514Q - - 1,22 9,71 E-06 2,2

S342Q - - 5,13 9,68E-06 2,2

V514I - - 2,02 9,20E-06 2,1

F513T - - 0,10 9,08E-06 2,0

V514F - - 0,13 8,65E-06 1,9 A343I - - 0,66 8,59E-06 1,9

F512M - - 5,40 8,48E-06 1,9

S460C - - 5,67 8,40E-06 1,9

A459G - - 0,42 7,63E-06 1,7

S342V - - 2,55 7,52E-06 1,7

A343M - - 0,86 7,14E-06 1,6

V514E - - 0,10 7,12E-06 1,6

F512H - - 0,58 6,86E-06 1,5

V514S - - 1,40 6,68E-06 1,5

A343P - - 5,80 6,67E-06 1,5

S342N - - 4,96 6,58E-06 1,5

S460V - - 4,80 6,51 E-06 1,5

S342I - - 1,34 6,23E-06 1,4

I267A - - 0,29 6,10E-06 1,4

S342C - - 5,07 5,57E-06 1,2

F513C - - 0,96 5,13E-06 1,2

F513A - - 0,54 5,02E-06 1,1

S342F - - 0,78 4,96E-06 1,1

S342C A343P - 3,03 4,64E-06 1,0

S342H A343P - 2,27 4,46E-06 1,0

S342M - - 4,57 4,36E-06 1,0

S342K A343P - 2,14 4,29E-06 1,0

I267V - - 0,81 4,23E-06 0,9

F512Q - - 0,68 4,05E-06 0,9

L458T - - 0,98 3,88E-06 0,9

S460I - - 6,53 3,69E-06 0,8

F515Y - - 0,74 1 ,67E-06 0,4

S461N - - 0,00 - -

T511V - - 0,00 - -

M463S - - 0,00 - -

T511A - - 0,00 - -

T511F - - 0,00 - -

F513Q - - 0,00 - -

F513S - - 0,00 - -

S461Q - - 0,00 - -

F515Q - - 0,00 - -

F515C - - 0,00 - -

T511C - - 0,00 - -

F515I - - 0,00 - -

M463V L544A - 0,00 - -

F512E - - 0,00 - -

T511N - - 0,00 - - F515H - - 0,00 - -

M463V L544C - 0,00 - -

L544E - - 0,00 - -

T511E - - 0,00 - -

F513E - - 0,00 - -

M463V L544T - 0,00 - -

F515N - - 0,00 - -

F515D - - 0,00 - -

T511F - - 0,00 - -

T511L - - 0,00 - -

F515T - - 0,00 - -

F515E - - 0,00 - -

V514Y - - 0,00 - -

F513D - - 0,00 - -

F515S - - 0,00 - -

F515L - - 0,00 - -

T511H - - 0,00 - -

F513H - - 0,00 - -

T511D - - 0,00 - -

T511I - - 0,00 - -

T511Q - - 0,00 - -

F515V - - 0,00 - -

T511M - - 0,00 - -

F513N - - 0,00 - -

M463V L544S - 0,00 - -

T511K - - 0,00 - -

I267F - - 0,06 - -

I267K - - 0,02 - -

I267M - - 0,37 - -

I267N - - 0,20 - -

I267R - - 0,02 - -

L458D - - 0,07 - -

L458F - - 0,04 - -

L458N - - 0,24 - -

A343L - - 0,02 - -

M463A - - 5,03 - -

M463C - - 6,05 - -

M463D - - 0,28 - -

A459V - - 0,01 - -

S461C - - 0,14 - -

S461D - - 0,00 - -

S461I - - 0,01 - - S461 T - - 0,77 - -

M463V - - 0,80 - -

F512I - - 1 ,13 - -

F512L - - 2,85 - -

F512N - - 0,10 - -

F512T - - 0,31 - -

F512V - - 0,63 - -

F512Y - - 0,67 - -

L544A - - 0,88 - -

L544D - - 0,16 - -

L544F - - 0,15 - -

L544M - - 0,80 - -

L544N - - 0,81 - -

L544S - - 0,85 - -

L544T - - 2,07 - -

I267W - - 0,08 - -

I267S - - 0,37 - -

S460D - - 2,30 - -

S461V - - 0,1 1 - -

F512S - - 0,03 - -

M463S - - 1 ,95 - -

M463T - - 0,00 - -

F512D - - 0,05 - -

Table 5

200μΙ Assay

Assay components: final concentrations volumes

2 mM Inhibitor in 100%DMSO 10 1 .0E-4 M

Mix A

50 mM Tris/HCI pH 7,7 84,7 50 mM

0,10% Cocarboxylase (TPP) 3,4 0,002%

10 mM β-NADH 10 0,5 mM

300 mM MgCI₂ 10 15 mM

10 units/ml (1 :100) a-GDH/TPI 3,4 0,17 units/ml

10 mM D-Ribose 5-Phosphat 15 0,75 mM

Transketolase enzyme concentration dependent

140μΙ

Reaction Start:

depends on

Xylulose-5-Phosphat

substrate

50 mM Tris/HCI pH 7,7 ad. 60 μΙ_ 50 mM

60 μΙ

Indicator Reaction

200μΙ

Assay Components: Final Concentrations

Volumes

Mix A

2mM Inhibitor 10 1 .0E-4 M

50mM Tris/HCI pH 7,7 Ad.140 50mM

0,10% Cocarboxylase (TPP) 3,4 0,002%

10 mM β-NADH 10 0,5 mM

300mM MgCI₂ 10 15mM

1 units/ml (1 :1000) a-GDH/TPI 10 0,05 units/mL

140μΙ

Reaction Start:

2,94 mM Glycerinaldehyd 3-Phosphat 51 0,75mM

50mM Tris/HCI pH 7,7 9

60μΙ

EXAMPLE 5: Demonstration of herbicide tolerance in a transient tobacco expression system

Transient expression of transketolase genes (e.g. SEQ ID NO: 1 , 10, 104, 130 and 156) are done as described previously (Voinnet O., et al., 2003, The Plant Journal 33, 949-956). In brief, for transient transformation of tobacco leaves or stable Arabidopsis thaliana transformation, Wildtype or mutated TKL sequences encoding TKL polypeptides comprising SEQ ID NO: 1 , 10, 104, 130 and 156, are synthesized (Life Technologies, USA) and cloned with standard cloning techniques as described in Sambrook et al. (Molecular cloning (2001 ) Cold Spring Harbor Laboratory Press) in a binary vector containing resistance marker gene cassette (AHAS) and TKL sequence in between ubiquitin promoter (PcUbi) and nopaline synthase terminator (NOS) sequence. In addition, a plastid targeting peptide (amino acid 1- 59 from swiss prot gene id FENR_SPIOL) with a linker sequence CSSAAA was fused to SEQ ID NO: 104, 130 and 156. Young leaves of Nicotiana benthamiana are infiltrated with transgenic Agrobacterium suspension (strain: pGV 2260, OD⁶⁰⁰ of 1.0) harbouring binary vector constructs with a transketolase gene. Two to three days after infiltration punches of leave discs (0.75 cm in diameter) are transferred to 6-well plates with medium containing herbicide of interest in different concentrations. Multi well plates are incubated in a growth chamber at 22°C, 75% relative humidity and 1 10 μιηοΙ Phot * nr² * s-¹ with 14 : 10 h light : dark photoperiod. Herbicide effect are monitored by visual inspection and analysis of photosynthetic yield by imaging PAM (Walz, Effeltrich, Germany) 24h, 48h and 96h after treatment. Tolerance factors are calculated based on IC50 values of PSIl yield inhibition of transformed versus empty vector-transformed leave discs. IC50 of PSIl yield inhibition in empty vector-transformed leaf discs treated with Cornexistin for 24h or 48 h was measured with 4.7*10-⁶ M or 1.0* 10 ⁶ M, respectively.

Table 6: Tolerance values as observed in transient expression system, 24h after treatment.

Gene Seq Cornexistin

ID

LEMPA TKL wt 10 2.9

SPIOL_TKL wt 1 3.6

Pd_TKL1 104 5.9

Pd_TKL2 130 2.8

Pd_TKL3 156 4.8

SPIOL TKL S461T 1* 4.5

SPIOL TKL L544T V 2.1

* with respective mutation as stated in column 'gene'

EXAMPLE 6: Maize whole plant transformation and herbicide tolerance testing: Immature embryos were transformed according to the procedure outlined in Peng et al.

(WO2006/136596). Plants were tested for the presence of the T-DNA by Taqman analysis with the target being the nos terminator which is present in all constructs. Healthy looking plants were sent to the greenhouse for hardening out and subsequent spray testing.

The plants were individually transplanted into MetroMix 360 soil in 4" pots. Once they had been in the greenhouse (day/night cycle of 27°C /21 °C with 14 hour day length supported by 600W high pressure sodium lights), they were allowed to grow for 14 days. They were then sprayed with a herbicide treatment. Herbicide injury evaluations were taken 7 and 14 days after treatment to look for injury to new growth points and overall plant health. The top survivors were transplanted into gallon pots filled with MetroMix 360 for seed production.

EXAMPLE 7: Soybean transformation and herbicide tolerance testing:

Soybean cv Jake was transformed as previously described¹. After regeneration, transformants were transplanted to soil in small pots, placed in growth chambers (16 hr day/ 8 hr night; 25°C day/ 23°C night; 65% relative humidity; 130-150 μΕ nr² s^_1) and subsequently tested for the presence of the T-DNA via Taqman analysis. After a few weeks, healthy, transgenic positive, single copy events were transplanted to larger pots and allowed to grow in the growth chamber. An optimal shoot for cutting was about 3-4" tall, with at least two nodes present. Each cutting was taken from the original transformant (mother plant) and dipped into rooting hormone powder (indole-3-butyric acid, IBA). The cutting was then placed in oasis wedges inside a bio- dome. The mother plant was taken to maturity in the greenhouse and harvested for seed. Wild type cuttings were also taken simultaneously to serve as negative controls. The cuttings were kept in the bio-dome for 5-7 days and then transplanted to 3" pots and then acclimated in the growth chamber for two more days. Subsequently, the cuttings were transferred to the greenhouse, acclimated for approximately 4 days, and then subjected to herbicide spray tests. Herbicide injury evaluations were taken at 7 and 14 days after treatment.

References

1 . Hong HP, Zhang H, Olhoft P, Hill S, Wiley H, Toren E, Hillebrand H, Jones T, Cheng M. Organogenic callus as the target for plant regeneration and transformation via Agrobacterium in soybean {Glycine max (L.) Merr.). In Vitro Cell. Dev. Biol.-Plant 2007: 43: 558-568

EXAMPLE 8. Engineering herbicide tolerant plants

Herbicide tolerant soybean (Glycine max) or corn (Zea mays) plants are generated as described by Olhoft et al. (US patent 2009/0049567). For transformation of soybean or Arabidopsis thaliana, transketolase genes are cloned with standard cloning techniques as described in Sambrook et al. (Molecular cloning (2001 ) Cold Spring Harbor Laboratory Press) in a binary vector containing resistance marker gene cassette (AHAS) and transketolase sequence (marked as GOI) in between ubiquitin promoter (PcUbi) and nopaline synthase terminator (NOS) sequence. For corn transformation, transketolase sequences are cloned with standard cloning techniques as described in Sambrook et al. (Molecular cloning (2001 ) Cold Spring Harbor Laboratory Press) in a binary vector containing resistance marker gene cassette (AHAS) and transketolase sequence (marked as GOI) in between corn ubiquitin promoter (ZmUbi) and nopaline synthase terminator (NOS) sequence. Binary plasmids are introduced to Agrobacterium tumefaciens for plant transformation. Plasmid constructs are introduced into soybean's axillary meristem cells at the primary node of seedling explants via Agrobacterium-mediated transformation. After inoculation and co-cultivation with Agrobacteria, the explants are transferred to shoot introduction media without selection for one week. The explants were subsequently transferred to a shoot induction medium with 1 -3 μΜ imazapyr (Arsenal) for 3 weeks to select for transformed cells. Explants with healthy callus/shoot pads at the primary node are then transferred to shoot elongation medium containing 1 -3 μΜ imazapyr until a shoot elongated or the explant died. Transgenic plantlets are rooted, subjected to TaqMan analysis for the presence of the transgene, transferred to soil and grown to maturity in greenhouse. Transformation of corn plants are done by a method described by McElver and Singh (WO 2008/124495). Plant transformation vector constructs containing transketolase sequences are introduced into maize immature embryos via Agrobacterium-mediated transformation.

Transformed cells are selected in selection media supplemented with 0.5-1 .5 μΜ imazethapyr for 3-4 weeks. Transgenic plantlets are regenerated on plant regeneration media and rooted afterwards. Transgenic plantlets are subjected to TaqMan analysis for the presence of the transgene before being transplanted to potting mixture and grown to maturity in greenhouse. Arabidopsis thaliana are transformed with transketolase sequences by floral dip method as described by McElver and Singh (WO 2008/124495). Transgenic Arabidopsis plants are subjected to TaqMan analysis for analysis of the number of integration loci. Transformation of Oryza sativa (rice) are done by protoplast transformation as described by Peng et al. (US 6653529) EXAMPLE 9: Demonstration of herbicide tolerance

TO or T1 transgenic plant of soybean, corn, and rice containing transketolase sequences are tested for improved tolerance to TK-inhibiting herbicides in greenhouse studies and mini-plot studies. For the pre-emergence treatment, the herbicides are applied directly after sowing by means of finely distributing nozzles. The containers are irrigated gently to promote germination and growth and subsequently covered with transparent plastic hoods until the plants have rooted. This cover causes uniform germination of the test plants, unless this has been impaired by the herbicides. For post emergence treatment, the test plants are first grown to a height of 3 to 15 cm, depending on the plant habit, and only then treated with the herbicides. For this purpose, the test plants are either sown directly and grown in the same containers, or they are first grown separately and transplanted into the test containers a few days prior to treatment.

For testing of TO plants, cuttings can be used. In the case of soybean plants, an optimal shoot for cutting is about 7.5 to 10 cm tall, with at least two nodes present. Each cutting is taken from the original transformant (mother plant) and dipped into rooting hormone powder (indole-3-butyric acid, IBA). The cutting is then placed in oasis wedges inside a bio-dome. Wild type cuttings are also taken simultaneously to serve as controls. The cuttings are kept in the bio-dome for 5-7 days and then transplanted to pots and then acclimated in the growth chamber for two more days. Subsequently, the cuttings are transferred to the greenhouse, acclimated for approximately 4 days, and then subjected to spray tests as indicated. Depending on the species, the plants are kept at 10-25°C or 20-35°C. The test period extends over 3 weeks. During this time, the plants are tended and their response to the individual treatments is evaluated. Herbicide injury evaluations are taken at 2 and 3 weeks after treatment. Plant injury is rated on a scale of 0% to 100%, 0% being no injury and 100% being complete death.

Transgenic Arabidopsis thaliana plants are assayed for improved tolerance to TK-inhibiting herbicides in 48-well plates. Therefore, T2 seeds are surface sterilized by stirring for 5 min in ethanol + water (70+30 by volume), rinsing one time with ethanol + water (70+30 by volume) and two times with sterile, deionized water. The seeds are resuspended in 0.1 % agar dissolved in water (w/v) Four to five seeds per well are plated on solid nutrient medium consisting of half-strength murashige skoog nutrient solution, pH 5.8 (Murashige and Skoog (1962) Physiologia Plantarum 15: 473-497). Compounds are dissolved in dimethylsulfoxid (DMSO) and added to the medium prior solidification (final DMSO concentration 0.1 %). Multi well plates are incubated in a growth chamber at 22°C, 75% relative humidity and 1 10 μιηοΙ Phot * nr² * s^_1 with 14 : 10 h light : dark photoperiod. Growth inhibition is evaluated seven to ten days after seeding in comparison to wild type plants (Table 6). Additionally, transgenic T1 Arabidopsis plants can be tested for improved tolerance to herbicides in greenhouse studies with TK-inhibiting herbicides such as cornexisthin.

Table 7: Relative tolerance rates of transgenic Arabidopsis plants as compared to a non- transgenic Arabidopsis plants (non-transgenic = 1.0), treated with Cornexistin. Growth inhibition is evaluated seven to ten days after seeding in comparison to wild type plants.

Gene Seq Cornexistin

ID

LEMPA_TKL wt 10 9

SPIOL_TKL wt 1 4.8

Pd_TKL1 104 28

Pd_TKL2 130 8.6

Pd_TKL3 156 10

EXAMPLE 10: Sequence Analysis.

Leaf tissue was collected from clonal plants separated for transplanting and analyzed as individuals. Genomic DNA was extracted using a Wizard® 96 Magnetic DNA Plant System kit (Promega, US Patent Nos. 6,027,945 & 6,368,800) as directed by the manufacturer. Isolated DNA was PCR amplified using the appropriate forward and reverse primer.

PCR amplification was performed using Hotstar Taq DNA Polymerase (Qiagen) using touchdown thermocycling program as follows: 96°C for 15 min, followed by 35 cycles (96°C, 30 sec; 58°C - 0.2 °C per cycle, 30 sec; 72°C, 3 min and 30 sec), 10 min at 72°C. PCR products were verified for concentration and fragment size via agarose gel electrophoresis. Dephosphorylated PCR products were analyzed by direct sequence using the PCR primers (DNA Landmarks, or Entelechon). Chromatogram trace files (.scf) were analyzed for mutation relative to the wild-type gene using Vector NTI Advance 10™

(Invitrogen). Based on sequence information, mutations were identified in several individuals. Sequence analysis was performed on the representative chromatograms and corresponding AlignX alignment with default settings and edited to call secondary peaks.

Claims

A plant or plant part comprising a polynucleotide encoding a wildtype or mutated transketolase (TK) polypeptide, the expression of said polynucleotide confers to the plant or plant part tolerance to TK-inhibiting herbicides.

The plant or plant part of claim 1 , wherein the polynucleotide encoding the wildtype or mutated TK polypeptide comprises the nucleic acid sequence set forth in SEQ ID NO: 182 or 183, or a homologue, variant or derivative thereof.

The plant or plant part of any of claims 1 to 2, wherein the wildtype or mutated TK polypeptide is a functional variant having, over the full-length of the variant, at least about 60%, illustratively, at least about 80%, 90%, 95%, 98%, 99% or more amino acid sequence identity to SEQ ID NO: 1 ,

2,

3,

4,

5,

6,

7,

8,

9,

10,

1 1 ,

12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100, 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 1 1 1 , 1 12, 113, 1 14, 115, 1 16, 1 17, 118, 1 19, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 , 152, 153, 154, 155, 156, 157, 158, 159, 160, 161 , 162, 163, 164, 165, 166, 167, 168, 169, 170, 171 , 172, 173, 174, 175, 176, 177, 178, 179, 180, or 181.

The plant or plant part of any of claims 1 to 3, wherein the mutated TK refers to a TK polypeptide comprising the sequence of SEQ ID NO: 1 , an orthologue, paralogue, or homologue thereof, wherein the amino acid sequence differs from the wildtype amino acid sequence at one or more positions corresponding to positions 265, 267, 337, 342, 343, 458, 459, 460, 461 , 463, 51 1 , 512, 513, 514, 515, 544 of SEQ ID NO: 1.

A seed capable of germination into a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

A plant cell of or capable of regenerating a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides, wherein the plant cell comprises the polynucleotide operably linked to a promoter.

A plant cell comprising a polynucleotide operably linked to a promoter operable in a cell, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

A plant product prepared from a plant or plant part comprising in at least some of its cells cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

A progeny or descendant plant derived from a plant comprising in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, wherein the progeny or descendant plant comprises in at least some of its cells the polynucleotide operably linked to the promoter, the expression of the wildtype or mutated TK polypeptide conferring to the progeny or descendant plant tolerance to the TK-inhibiting herbicides.

A method for controlling weeds at a locus for growth of a plant, the method comprising: (a) applying an herbicide composition comprising TK-inhibiting herbicides to the locus; and (b) planting a seed at the locus, wherein the seed is capable of producing a plant that comprises in at least some of its cells a polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

The method of claim 10, wherein herbicide composition is applied to the weeds and to the plant produced by the seed.

A method of producing a plant having tolerance to TK-inhibiting herbicide, the method comprising regenerating a plant from a plant cell transformed with transformed with a recombinant polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

13. A method of producing a progeny plant having tolerance to TK-inhibiting herbicide, the method comprising: crossing a first TK-inhibiting herbicide-tolerant plant with a second plant to produce a TK-inhibiting herbicide- tolerant progeny plant, wherein the first plant and the progeny plant comprise in at least some of their cells a polynucleo- tide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides.

14. A plant or plant part comprising in at least some of its cells a recombinant polynucleotide operably linked to a promoter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides, wherein the plant or plant part further exhibits a second or third herbicide-tolerant trait.

15. A method for producing a plant product from the plant of claim 1 , the method comprising processing the plant or a plant part thereof to obtain the plant product.

16. The method of claim 15, wherein the plant product is fodder, seed meal, oil, or seed- treatment-coated seeds.

17. A plant product obtained from a plant or plant part thereof, wherein the plant or plant part comprises in at least some of its cells a polynucleotide operably linked to a pro- moter operable in plant cells, the promoter capable of expressing a wildtype or mutated TK polypeptide encoded by the polynucleotide, the expression of the wildtype or mutated TK polypeptide conferring to the plant tolerance to TK-inhibiting herbicides, wherein the plant or plant part further exhibits a second or third herbicide-tolerant trait.

18. The plant product of claim 17, wherein the product is fodder, seed meal, oil, or seed- treatment-coated seed.