WO2014132141A2 - Plantes tolérantes aux herbicides auxiniques - Google Patents

Plantes tolérantes aux herbicides auxiniques Download PDF

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Publication number
WO2014132141A2
WO2014132141A2 PCT/IB2014/001106 IB2014001106W WO2014132141A2 WO 2014132141 A2 WO2014132141 A2 WO 2014132141A2 IB 2014001106 W IB2014001106 W IB 2014001106W WO 2014132141 A2 WO2014132141 A2 WO 2014132141A2
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WIPO (PCT)
Prior art keywords
plant
herbicide
seq
auxinic
seed
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PCT/IB2014/001106
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English (en)
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WO2014132141A3 (fr
Inventor
J. Christopher Hall
Mithila JUGULAM
Scots Llewellyn MANKIN
Brigette J. WESTON
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Basf Se
University Of Guelph
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Publication of WO2014132141A2 publication Critical patent/WO2014132141A2/fr
Publication of WO2014132141A3 publication Critical patent/WO2014132141A3/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/12—Processes for modifying agronomic input traits, e.g. crop yield
    • A01H1/129—Processes for modifying agronomic input traits, e.g. crop yield involving hormone-influenced development, e.g. auxin
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/10—Seeds
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/20—Brassicaceae, e.g. canola, broccoli or rucola
    • A01H6/202—Brassica napus [canola]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance

Definitions

  • This application relates to the field of agriculture, particularly to auxinic herbicide - tolerant plants.
  • Auxinic herbicide tolerance has been reported as occurring in weeds, such as Kochia and Charlock, but not in Brassica crop plants, such as domesticated B. napus, B. rapa, or B. juncea. See, e.g., Nandula and Mathey (2002); Jugulam et al. (2005).
  • auxinic herbicide tolerant crop plant having a commercial level of tolerance to an auxinic herbicide.
  • Another aspect described herein is a non- transgenic auxinic herbicide tolerant crop plant having a commercial level of tolerance to an auxinic herbicide.
  • auxinic herbicide tolerant plant or plant part thereof having the auxinic herbicide-tolerance characteristic of any of Sinapis arvensis lines DT- 01 SA2-R or DT-01 BC8SA2-R, a representative sample of seed of each line having been deposited with American Type Culture Collection (ATCC) under Patent Deposit Designation Numbers PTA-11213 and PTA-11214, respectively, with the proviso that the plant is a monocot or dicot species other than Sinapis arvensis.
  • ATCC American Type Culture Collection
  • PTA-11213 and PTA-11214 Patent Deposit Designation Numbers PTA-11213 and PTA-11214
  • the auxinic herbicide tolerance trait is referred to as "DART.”
  • Lines DT-01 SA2-R and DT-01 BC8SA2-R also are known as "SaParR" and "SaBC8R,” respectively.
  • nucleic acid comprising: (a) a chimeric polynucleotide comprising both Sinapis arvensis nucleotide sequence and Brassica nucleotide sequence, wherein said polynucleotide encodes the DART trait of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT- 01 BC8SA2-R, a representative sample of seed of each line having been deposited with American Type Culture Collection (ATCC) under Patent Deposit Designation Numbers PTA- 120132, PTA-11211, PTA-12050, PTA-11212, PTA-11213, and PTA-11214, respectively; or (b) a mutagenized or recombinant polynucleotide encoding the DART trait of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13,
  • the DART trait is encoded by the Sinapis arvensis polynucleotide.
  • the nucleic acid encodes a functional DART trait, said nucleic acid comprising (a) a nucleotide sequences according to any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80; or (b) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence according to any one of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81, respectively; or a mature form thereof.
  • a nucleotide sequence encoding the amino acid sequence is at least 90% homologous to any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, or a synonymous codon substituted variant thereof.
  • the encoded amino acid sequence is 70%> homologous to any one of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81, or a mature form thereof.
  • the polynucleotide is isolated.
  • the nucleic acid is operably linked to a promoter operable in plant cells capable of expressing the polypeptide encoded by the nucleic acid, the expression of the polypeptide conferring to the plant or cell tolerance to an auxinic herbicide.
  • auxinic herbicide tolerant plant or plant part thereof having the auxinic herbicide-tolerance characteristic of any of Brassica napus lines DT- 01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, or DT-01 BC5Bn#13-l-18, a representative sample of seed of each line having been deposited with American Type Culture Collection (ATCC) under Patent Deposit Designation Numbers PTA-120132, PTA-11211, PTA- 12050, and PTA-11212, respectively, with the proviso that the plant is a monocot or dicot species other than Sinapis arvensis; and wherein the auxinic herbicide tolerance is greater than that exhibited by a wild type variety of said plant lacking said auxinic herbicide tolerance.
  • ATCC American Type Culture Collection
  • Introgressed Brassica napus lines containing an auxinic herbicide tolerant trait include DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, and DT-01 BC5Bn#13-l-18.
  • the auxinic herbicide tolerance trait present in any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R is referred to as "DART.”
  • the auxinic herbicide tolerant plant is a Brassica juncea, B. napus, or B. rapa plant.
  • the auxinic herbicide tolerant plant is a canola plant.
  • Another embodiment is a descendant of the auxinic herbicide tolerant plant wherein the descendant has the auxinic herbicide -tolerance characteristic.
  • the auxinic herbicide-tolerance characteristic is DART.
  • Another embodiment described herein is a seed of or capable of producing a plant having the auxinic herbicide-tolerance characteristic of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R, with the proviso that the seed is other than a Sinapis arvensis seed.
  • the auxinic herbicide-tolerance characteristic is DART.
  • Another embodiment is a seed of or capable of producing a plant with the auxinic herbicide-tolerance characteristic.
  • the auxinic herbicide-tolerance characteristic is DART.
  • the seed has, disposed on a surface thereof, a composition comprising at least one agronomically acceptable ingredient.
  • the ingredient is at least one agronomically acceptable herbicide, fungicide, nematicide, or insecticide, or a combination thereof.
  • the insecticide comprises at least one anti-coleopteran agent, anti- hemipteran agent, anti-lepidopteran agent, or a combination thereof.
  • Another embodiment is a method for treating a seed with an agronomically acceptable composition.
  • the method comprises contacting the seed with an agronomically acceptable composition.
  • the agronomically acceptable composition comprises an auxinic herbicide.
  • Another embodiment described herein is a cell of a plant having the auxinic herbicide- tolerance characteristic of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R, with the proviso that the cell is of a monocot or dicot species other than Sinapis arvensis.
  • the auxinic herbicide -tolerance characteristic is DART.
  • plant product produced from a plant having the auxinic herbicide-tolerance characteristic of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R.
  • a plant product of a plant comprising the auxinic herbicide-tolerance characteristic is DART.
  • plant products include, but are not limited to, seeds, hulled or dehulled grain, oil, and meal.
  • plant products include whole plant, aerial plant part, stem, and/or leaf feed, fodder, and forages.
  • the plant product is grain, meal, or oil.
  • One embodiment described herein is a method for controlling weeds at a locus for growth of a plant.
  • the locus for growth of a plant is a field.
  • the method comprises: applying a composition comprising an auxinic herbicide to the locus, wherein the plant has an auxinic herbicide -tolerance characteristic of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R.
  • the auxinic herbicide-tolerance characteristic is the DART trait.
  • Another embodiment described herein is a method for controlling weeds in a field by application of an auxinic herbicide without significantly inhibiting the growth of a Brassica plant, the method comprising: (a) providing a Brassica plant or seed of any one of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R; and (b) applying an herbicide composition comprising an effective amount of an auxinic herbicide: (i) to the field, followed by planting of said plant or seed in therein; (ii) to the field, during or after planting of said seed therein; (iii) to the plant in said field and to weeds in the vicinity of the plant; (iv) to said seed, followed by planting of said seed in the field; or (v) to a plant by the seed after it has been planted in the field, and to weeds in the vicinity of the plant;
  • the plant is a Brassica juncea, B. napus, or B. rapa plant.
  • the plant is a canola plant.
  • the step of applying comprises performing post-emergent treatment of the plant by applying an herbicide composition, comprising auxinic herbicide(s), to the plant and its immediate vicinity, at a dose rate of about 10 to about 5000 grams active ingredient per hectare (ai/ha).
  • the step of applying comprises performing pre-emergent treatment, or 0 to 30 days-pre-planting treatment, of the plant by applying an herbicide composition, comprising auxinic herbicide(s), to the seed planting locus thereof and its immediate vicinity, at a dose rate of about 10 to about 5000 g ai/ha.
  • Another embodiment described herein is a method for controlling weeds in a crop field by use of an auxinic herbicide without significantly inhibiting the growth of a crop plant, the method comprising: (a) providing a seed-treatment-treated seed comprising the nucleic acid of claim 41, the expression of said nucleic acid conferring to the plant or seed tolerance to an auxinic herbicide, the seed treatment comprising an auxinic herbicide; and (b) planting said treated seed in the field.
  • Another embodiment described herein is a method for producing an auxinic herbicide- tolerant progeny plant.
  • the method comprises: crossing the parent plant with an auxinic herbicide tolerant plant having the auxinic herbicide-tolerance characteristic of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT- 01 SA2-R, or DT-01 BC8SA2-R, with the proviso that the plant is a monocot or dicot species other than Sinapis arvensis, to introduce the auxinic herbicide -tolerance characteristic into the germplasm of the progeny plant, wherein the progeny plant has increased tolerance to the auxinic herbicide relative to the parent plant.
  • Another embodiment described herein is a method for producing an auxinic herbicide- tolerant progeny plant.
  • the method comprises: (a) providing a parent plant of a desired line; and (b) crossing the parent plant with any one of the plants according of claims 1-4 to introduce the auxinic herbicide-tolerance characteristic into the germplasm of the progeny plant, wherein the progeny plant thereby has increased tolerance to an auxinic herbicide relative to the parent plant.
  • the method further comprises introgressing the auxinic herbicide-tolerance characteristic of the progeny plant through traditional plant breeding techniques to obtain a descendent plant having the auxinic herbicide-tolerance characteristic.
  • the parent plant comprises at least one herbicide tolerant (HT) mutant AHASL gene.
  • the parent plant is a dicot. In another aspect, the parent plant is a Brassica or Rhaphanus plant. In another aspect, the parent plant is a Brassica juncea, B. napus, or B. rapa plant. In yet another aspect, the parent plant is a canola plant.
  • Another embodiment described herein is a method for identifying an auxinic herbicide tolerant plant, or plant part thereof.
  • the method comprises: (a) providing biological material from a plant comprising the DART trait; (b) performing PCR, hybridization testing, or sequencing of said nucleic acid in said biological material to determine if said plant comprises the DART trait; and (c) identifying, based on the results of step (b), that the plant comprises the DART trait.
  • the plant comprises: (i) any one of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT- 01 BC8SA2-R, a representative sample of seed of each line having been deposited with American Type Culture Collection (ATCC) under Patent Deposit Designation Numbers PTA- 120132, PTA-11211, PTA-12050, PTA-11212, PTA-11213, and PTA-11214, respectively; (ii) a mutant, recombinant, or a genetically engineered derivative of any one of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT- 01 BC8SA2-R and that expressed the DART trait thereof; or (iii) a plant that is a progeny of at
  • Figure 1 Photographic results of an in vitro, whole plant dicamba kill curve.
  • Figure 2 Photographic images of a trichome trait observed in parent lines and Fl hybrid plant.
  • Figure 3 Dendrogram generated from SNP analysis.
  • Figure 4 Backcrossing scheme illustrating a procedure for development of wild mustard near-isogenic-lines (NILs) with auxinic herbicide-R (resistance) and herbicide-S (susceptibility).
  • NILs near-isogenic-lines
  • auxinic herbicide-R resistance
  • herbicide-S susceptibility
  • FIG. 5 Photographic images of wild mustard plants 21 days after planting (DAP). A and B indicate S and R plants, respectively.
  • Figure 6 Bar chart presenting leaf area (cm 2 ) of first true leaf of wild mustard (31 DAP).
  • S Parental-S; R: Parental-R; BC-Pl and BC-P2 are BC 4 F 1 progeny with larger leaf area and smaller leaf area, respectively.
  • Vertical bars represent SEM (Standard Error of the Means).
  • Figure 7 Nucleotide sequence of markers 1-6 (M1-M6) (SEQ ID NOs: l-6, respectively).
  • Figure 8 Linkage map of wild mustard with markers 1-6 (M1-M6).
  • R Auxinic herbicide resistance genetic locus.
  • Figure 9 Ovule/embryo rescue procedure to produce hybrids between B. juncea, B. rapa, and S. arvensis.
  • A represents the immature silique cultured on media (A or B);
  • B represents ovule excised from the siliques and
  • C denotes the hybrid plant regeneration form the ovule.
  • Figure 10 Production of hybrids between B. juncea, B. rapa, and S. arvensis.
  • A, B, C represent B. juncea, B. rapa, and S. arvensis, respectively.
  • D, Dl, and E, El illustrate hybrids produced via embryo rescue from crosses between B. juncea x S. arvensis and B. rapa x S. arvensis, respectively. Note, the hybrids exhibiting intermediate characteristics of the parents.
  • Figure 11 Plant response to dicamba, 200 g acid equivalents/hectare (ae/ha), 5 weeks after treatment.
  • A, B, C represent B. juncea, B. rapa, and S. arvensis, respectively.
  • D and E indicate hybrids produced by crossing B. juncea x S. arvensis and B. rapa x S. arvensis, respectively.
  • tolerant indicates a plant or plant part thereof capable of growing in the presence of an amount of herbicide that normally causes growth inhibition or phytotoxicity in a non-herbicide-tolerant (e.g., a wild-type) plant or portion thereof.
  • 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 quantity of herbicide or rate of application recommended by herbicide manufacturers. The maximum level or rate of herbicide application is the amount of herbicide that would normally inhibit the growth or cause phytotoxicity of a non-herbicide tolerant plant (e.g., a wild type plant or weed).
  • the terms “herbicide-tolerant” and “herbicide -resistant” are used interchangeably and are intended to have an equivalent meaning and an equivalent scope.
  • the terms “herbicide-tolerance” and “herbicide-resistance” are used interchangeably and are intended to have an equivalent meaning and an equivalent scope.
  • the terms “tolerant” and “resistant” are used interchangeably and are intended to have an equivalent meaning and an equivalent scope.
  • the phrases "auxinic herbicide tolerance trait” or “auxinic herbicide tolerance characteristic” are used interchangeably and are intended to have an equivalent meaning and an equivalent scope.
  • the auxinic herbicide tolerance trait is the auxinic herbicide tolerance phenotype present in any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R, and is referred to herein as "DART.”
  • the auxinic herbicide tolerance trait is effected by a functional expression product of a nucleic acid comprising the nucleotide sequence of any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, or a synonymous codon substituted variant thereof.
  • the expression product may be a polypeptide comprising the amino acid sequence of any one of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81, or a mature form thereof, or a conservatively substituted variant thereof.
  • a nucleotide sequence encoding the amino acid sequence is at least 90% homologous to any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, or a synonymous codon substituted variant thereof.
  • the encoded amino acid sequence is 70% homologous to any one of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81, or a mature form thereof.
  • the phrases "without inhibiting the growth” or “without significantly inhibiting the growth” are used to mean that a dicamba-treated plant comprising the DART trait will exhibit a dicamba tolerance of 75% or more, as determined 5 weeks after treatment with an effective amount of the herbicide. Yet, that amount of dicamba will inhibit the growth of, or can even result in the death of, a wild type variety of the plant, i.e. one lacking the DART trait.
  • auxinic herbicide As used herein in regard to herbicides useful in various embodiments hereof, terms such as auxinic herbicide, AHAS inhibitor, acetyl-Coenzyme A carboxylase (ACCase) inhibitor, PPO inhibitor, EPSPS inhibitor, imidazolinone, sulfonylurea, and the like, refer to those agronomically acceptable herbicide active ingredients (A.I.s) recognized in the art. Similarly, terms such as fungicide, nematicide, pesticide, and the like, refer to other agronomically acceptable A.I.s recognized in the art.
  • auxinic herbicide As used herein in regard to herbicides useful in various embodiments hereof, terms such as auxinic herbicide, AHAS inhibitor, acetyl-Coenzyme A carboxylase (ACCase) inhibitor, PPO inhibitor, EPSPS inhibitor, imidazolinone, sulfonylurea, and the like, refer to those agronomically
  • herbicide tolerant When used in reference to a particular mutant enzyme or polypeptide, terms such as herbicide tolerant (HT) 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.
  • herbicide tolerant when used specifically with regard to an AHAS enzyme, or AHASL polypeptide, it refers specifically to the ability to tolerate an AHAS-inhibitor.
  • Classes of AHAS -inhibitors include sulfonylureas, imidazolinones, triazolopyrimidines, sulfonylaminocarbonyltriazolinones, and pyrimidinyloxy[thio]benzoates.
  • the AHASL is preferably one that comprises at least one herbicide tolerance mutation located at amino acid residue position 122, 205, 574, or 653 ⁇ Arabidopsis thaliana AHASL numbering); and in some embodiments in which a sulfonylurea herbicide is to be used, the AHASL is preferably one that comprises at least one herbicide tolerance mutation located at amino acid residue position 197 or 574 ⁇ Arabidopsis thaliana AHASL numbering).
  • 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 its natural context 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.
  • transgenic plant refers to a plant that comprises a heterologous polynucleotide that has been inserted by the use of a recombinant DNA technique.
  • 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 the heterologous nucleic acid introduced by the use of a recombinant DNA technique, 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.
  • a transgenic organism can also be referred to herein as a "recombinant" organism.
  • 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.
  • 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 a non-recombinant technique through human action. Any mutagenesis 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.
  • mutagenesis followed by selection with an herbicide (e.g., auxinic herbicides), in order to obtain a mutagenized organism having an herbicide tolerance trait or DNA encoding such trait, include, but are not limited to: tissue culture of plant cells (e.g., calli) to induce tissue culture mutagenesis, treatment of plant cells with a chemical mutagen, or treatment of plant cells with radiation (e.g., gamma rays, X-rays, or subatomic particles).
  • tissue culture of plant cells e.g., calli
  • treatment of plant cells with a chemical mutagen e.g., gamma rays, X-rays, or subatomic particles.
  • chimeric biomolecule is a non-naturally-occurring biomolecule that contains sequence segments from two different species, attached to each other.
  • a non-naturally-occurring, chimeric DNA can be prepared by artificially cross- pollinating plants of different species.
  • a chimeric nucleic acid can comprise nucleotide sequence from a first species attached to a nucleotide sequence from a second species. The different sequence segments are directly attached one to another.
  • a Sinapis arvensis polynucleotide segment attached to a Brassica polynucleotide segment will together constitute a chimeric nucleic acid.
  • chimeric polypeptides can also be formed by expression of a chimeric nucleic acid coding sequence.
  • a non-transgenic, non- naturally-occurring, chimeric DNA prepared by artificially cross-pollinating plants of different species, is one that in nature is not transmitted to progeny without having been artificially propagated at its inception, e.g., by embryo rescue and/or by tissue culture and plantlet regeneration.
  • GMO genetically modified organism
  • 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).
  • GMO plant can contain bacterial genetic material
  • GMO plant can contain genetic material from another plant.
  • "recombinant,” “transgenic,” and “GMO” are considered synonyms and indicate the presence of genetic material from a different source.
  • 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.
  • progeny refers to a first generation plant.
  • seed comprises seeds of all types, such as, for example, true seeds, caryopses, achenes, fruits, tubers, seedlings and similar forms.
  • seed refers to true seed(s) unless otherwise specified.
  • the seed used can be seed of the useful plants mentioned above, but also the seed of transgenic plants or plants obtained by traditional breeding methods. Examples of traditional breeding methods can include crossbreeding, selfmg, back-crossing, embryo rescue, in-crossing, out-crossing, inbreeding, selection, asexual propagation, and other traditional techniques as are known in the art.
  • the term "derived from,” unless otherwise specified, indicates that a particular thing (e.g., plant, seed, etc.) or group of things has originated from the source specified, but has not necessarily been obtained directly from the specified source.
  • auxinic herbicides can be employed with a variety of commercially valuable plants including, but not limited to, auxinic herbicide- tolerant Brassica species, such as B. oleracea, B. rapa, B. nigra, and B. juncea.
  • auxinic herbicide-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 auxinic herbicide-tolerance trait donor lines for development, as by traditional plant breeding, to produce other varietal and/or hybrid crops containing such trait.
  • All such resulting variety or hybrids crops, containing the ancestral auxinic herbicide-tolerance trait can be referred to herein as progeny of the ancestral, auxinic herbicide-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.
  • Brassica A-, B-, and C-genome-located auxinic herbicide traits these can be bred into Brassica species having a corresponding genome, e.g., B. napus (AACC), B. juncea (AABB), B. oleracea (CC), B. rapa (AA), B. nigra (BB), B. carinata (BBCC), and Raphanobrassica varieties that are progeny of a cross between any of the foregoing and a Raphanus spp., e.g., Raphanobrassica var. 'rabbage' (RRCC) from B.
  • AACC B. napus
  • AABB B. oleracea
  • AA B. rapa
  • BBCC B. nigra
  • BBCC B. carinata
  • Raphanobrassica varieties that are progeny of a cross between any of the foregoing and a Raphanus spp., e
  • the auxinic herbicide-tolerant plants described herein can be employed as auxinic herbicide-tolerance characteristic donor lines to produce other varietal and/or hybrid crops containing such a characteristic.
  • the selected Brassica will be a canola variety.
  • Non-limiting examples of Brassica taxa that can provide useful canola varieties include B. rapa (esp.
  • a "canola” plant, line, or variety is a Brassica plant, line, or variety that has been bred or otherwise engineered to produce seed oil that, compared to seed oil of members of the parent line or of a wild-type line, is reduced in erucic acid content, or in both erucic acid and glucosinolate content.
  • the canola will be a food-grade canola, i.e., wherein: (1) the seed oil thereof contains 2% by weight or less of its fatty acids as erucic acid, preferably less than or about 1%, 0.5% or, 0.2% thereof, or essentially 0%> thereof; and (2) the oil-removed, dry seed meal contains less than 30 micromoles per gram of any one or any mixture of 3-butenyl glucosinolate, 2-hydroxy-3-butenyl glucosinolate, 4-pentenyl glucosinolate, and 2-hydroxy-4- pentenyl glucosinolate, preferably less than or about 20, 15, 10, 5, or 2 umol/g thereof, or essentially 0 ⁇ /g thereof.
  • the canola plant, line, or variety is one in which the indole-glucosinolate content, e.g., the content of glucobrassicin (3-indolylmethyl- glucosinolate), 4-hydroxy-glucobrassicin, 4-methoxy-glucobrassicin, and/or neoglucobrassicin (1-methoxy-glucobrassicin), in the seed meal is modified compared to that in seed meal of the parent plant or a wild-type, line, or variety; in some embodiments, such a canola can also be a food-grade canola.
  • the indole-glucosinolate content e.g., the content of glucobrassicin (3-indolylmethyl- glucosinolate), 4-hydroxy-glucobrassicin, 4-methoxy-glucobrassicin, and/or neoglucobrassicin (1
  • canola can appear per se or in combination terms such as “canola/OSR” and “canola/oilseed rape;” the combined term referring to oilseed Brassica canola varieties.
  • the term “oilseed rape,” when not modified by combination with the term “canola” is understood as encompassing both canola and non-canola varieties of oilseed Brassicas, i.e., Brassicas that express useful seed oils. Seed oils can be removed from the seed by crushing, pressing, solvent extraction, any other techniques known useful in the art.
  • One embodiment described herein is an auxinic herbicide -tolerant plant or plant part thereof, wherein the plant is a plant of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT- 01 BC4Bn#13-l, or DT-01 BC5Bn#13-l-18.
  • the herbicide-tolerance characteristic of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, and DT-01 BC5Bn#13-l-18 are the same herbicide tolerance-characteristic.
  • the herbicide-tolerance characteristic of lines DT-01 SA2-R and DT-01 BC8SA2-R are the same herbicide tolerance-characteristic.
  • the herbicide-tolerance characteristic is DART.
  • a deposit of a representative sample of seeds of each of lines DT-01 BC3Bn#13, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R was made by BASF Plant Science, 26 Davis Drive, Research Triangle Park, NC 27709, United States of America and University of Guelph, Unit 102, 150 Research Lane, Guelph, Ontario, NIG 4T2, Canada with the ATCC, 10801 University Boulevard., Manassas, Virginia, 20110, United States of America on July 16, 2010; a deposit of a representative sample of seeds of line DT-01 BC4Bn#13-l was made by BASF Plant Science and University of Guelph with the ATCC on August 30, 2011; and a deposit of a representative sample of seeds of line DT-01 Cyc2 BNS4 was made by BASF Plant Science and University of Guelph with the ATCC on January 15, 2013.
  • the nucleic acid is isolated.
  • the nucleic acid comprises (a) a nucleotide sequence according to any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80; or (b) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence according to any one of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81, respectively; or a mature form thereof.
  • the nucleic acid comprises: (a) the nucleotide sequence of SEQ ID NO: 8; (b) a nucleotide sequence encoding a translation elongation factor EF1 A/initiation factor family polypeptide comprising the amino acid sequence of SEQ ID NO:9, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO: 8 and encodes an auxinic-herbicide-tolerant, functional, translation elongation factor EF1 A/initiation factor family polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, translation elongation factor EF1 A/initiation factor family polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:9.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO: 16; (b) a nucleotide sequence encoding a calmodulin 5-like polypeptide comprising an amino acid sequence according to SEQ ID NO: 17, or a mature form thereof; (c) a nucleotide sequence that is at least 90% homologous to SEQ ID NO: 16 and encodes an auxinic- herbicide-tolerant, functional, calmodulin 5-like polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, calmodulin 5-like polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO: 17.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO: 18; (b) a nucleotide sequence encoding an ARF21 transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO: 19, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO: 18 and encodes an auxinic-herbicide-tolerant, functional, ARF21 transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, ARF21 transcription factor polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO: 19.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:22; (b) a nucleotide sequence encoding a TPR3 transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO:23, or a mature form thereof; (c) a nucleotide sequence that is at least 90% homologous to SEQ ID NO:22 and encodes an auxinic- herbicide-tolerant, functional, TPR3 transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, TPR3 transcription factor polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:23.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:30; (b) a nucleotide sequence encoding a Endo-Mitochondrial, Auxin-Associated Protein- 1 (mtAAP-1) polypeptide comprising an amino acid sequence according to SEQ ID NO:31, or a mature form thereof; (c) a nucleotide sequence that is at least 90% homologous to SEQ ID NO:30 and encodes an auxinic-herbicide-tolerant, functional, Endo-Mitochondrial, Auxin- Associated Protein- 1 (mtAAP-1) polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, Endo-Mitochondrial, Auxin- Associated Protein- 1 (mtAAP-1) polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:31.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:36; (b) a nucleotide sequence encoding a SAUR-like auxin-responsive protein family transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO:37, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO:36 and encodes an auxinic-herbicide-tolerant, functional, SAUR-like auxin- responsive protein family transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, SAUR-like auxin-responsive protein family transcription factor polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:37.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:40; (b) a nucleotide sequence encoding a HTA13 histone polypeptide comprising an amino acid sequence according to SEQ ID NO:41, or a mature form thereof; (c) a nucleotide sequence that is at least 90% homologous to SEQ ID NO:40 and encodes an auxinic-herbicide- tolerant, functional, HTA13 histone polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, HTA13 histone polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:41.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:44; (b) a nucleotide sequence encoding a Knotted 1 like (KNAT4) transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO:45, or a mature form thereof; (c) a nucleotide sequence that is at least 90% homologous to SEQ ID NO:44 and encodes an auxinic-herbicide-tolerant, functional, Knotted 1 like (KNAT4) transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, Knotted 1 like (KNAT4) transcription factor polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:45.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:48; (b) a nucleotide sequence encoding a Zinc-Finger-Like, Auxin-Associated Protein- 1 (zFAAP-1) polypeptide comprising an amino acid sequence according to SEQ ID NO:49, or a mature form thereof; (c) a nucleotide sequence that is at least 90% homologous to SEQ ID NO:48 and encodes an auxinic-herbicide-tolerant, functional, Zinc-Finger-Like, Auxin- Associated Protein- 1 (zFAAP-1) polypeptide; or (d) a nucleotide sequence encoding an auxinic- herbicide-tolerant, functional, Zinc-Finger-Like, Auxin- Associated Protein- 1 (zFAAP-1) polypeptide comprising an amino acid sequence at least 70%> identical to SEQ ID NO:49.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:52; (b) a nucleotide sequence encoding a Alba DNA/RNA-binding protein transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO:53, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO:52 and encodes an auxinic-herbicide-tolerant, functional, Alba DNA/RNA-binding protein transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic- herbicide-tolerant, functional, Alba DNA/RNA-binding protein transcription factor polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:53.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:56; (b) a nucleotide sequence encoding an IAA16 transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO:57, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO:56 and encodes an auxinic- herbicide-tolerant, functional, IAA16 transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, IAA16 transcription factor polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:57.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:62; (b) a nucleotide sequence encoding an IAA12 bodenlos/monopteros transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO: 63, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO: 62 and encodes an auxinic-herbicide-tolerant, functional, IAA12 bodenlos/monopteros transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, IAA12 bodenlos/monopteros transcription factor polypeptide comprising an amino acid sequence at least 70%> identical to SEQ ID NO: 63.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:66; (b) a nucleotide sequence encoding an RUB1 polypeptide comprising an amino acid sequence according to SEQ ID NO:67, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO: 66 and encodes an auxinic-herbicide- tolerant, functional, RUB1 polypeptide; or (d) a nucleotide sequence encoding an auxinic- herbicide-tolerant, functional, RUB1 polypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:67.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:76; (b) a nucleotide sequence encoding a CAM7 transcription factor polypeptide comprising an amino acid sequence according to SEQ ID NO:77, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO: 76 and encodes an auxinic- herbicide-tolerant, functional, CAM7 transcription factor polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, CAM7 transcription factor polypeptide comprising an amino acid sequence at least 70%> identical to SEQ ID NO: 77.
  • the nucleic acid comprises: (a) a nucleotide sequence according to SEQ ID NO:80; (b) a nucleotide sequence encoding a cytochrome P450 CYP83A1 polypeptide comprising an amino acid sequence according to SEQ ID NO:81, or a mature form thereof; (c) a nucleotide sequence that is at least 90%> homologous to SEQ ID NO: 80 and encodes an auxinic-herbicide-tolerant, functional, cytochrome P450 CYP83A1 polypeptide; or (d) a nucleotide sequence encoding an auxinic-herbicide-tolerant, functional, cytochrome P450 CYP83Alpolypeptide comprising an amino acid sequence at least 70% identical to SEQ ID NO:81.
  • the auxinic herbicide -tolerant plant is a descendent of a member of any one of DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l- 18, DT-01 SA2-R, or DT-01 BC8SA2-R.
  • the descendant was obtained by traditional plant breeding from said member.
  • One aspect described herein is a progeny or a descendant of an auxinic herbicide-tolerant plant, as well as seeds derived from the auxinic herbicide-tolerant plants, and cells derived from the auxinic herbicide-tolerant plants.
  • auxinic herbicide-tolerant plant lines described as useful herein can be employed as auxinic herbicide -tolerance characteristic(s) donor lines for development, as by traditional plant breeding, to produce other varietal and/or hybrid crops containing such trait. All such resulting variety or hybrids crops, containing the ancestral auxinic herbicide-tolerance characteristic or characteristic can be referred to herein as progeny or descendant of the ancestral, auxinic herbicide-tolerant line(s).
  • One embodiment described herein is a method for producing an auxinic herbicide- tolerant progeny plant, the method comprising: crossing a parent plant with an auxinic herbicide- tolerant plant having an auxinic herbicide -tolerance characteristic to introduce the auxinic herbicide-tolerance characteristic into the germplasm of the progeny plant, wherein the progeny plant has increased tolerance to the auxinic herbicide relative to the parent plant.
  • the auxinic herbicide -tolerant plant that is crossed with the parent plant is a plant of line DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R.
  • the method further comprises the step of introgressing the auxinic herbicide-tolerance characteristic of the progeny plant through traditional plant breeding techniques to obtain a descendent plant having the auxinic herbicide-tolerance characteristic.
  • a progeny plant is an interspecies crossed between different Brassica species.
  • Another example of a progeny plant is an intergeneric cross between Raphanus sativus with Brassica oleracea to provide a Raphanobrassica.
  • B. napus, B. rapa, and B. juncea are of particular interest, with B. napus being preferred in other embodiments.
  • auxinic herbicide-tolerant plants which, in addition to being auxinic herbicide-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; bleaching herbicides such as hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors or phytoene desaturase (PDS) inhibitors; enolpyruvyl shikimate 3-phosphate synthase (EPSPS) inhibitors such as glyphosate; glutamine synthetase (GS) inhibitors such as glufosinate; protoporphyrinogen-IX oxidase (PPO) inhibitors; lipid biosynthesis inhibitors such as ACCase inhibitors; or oxynil (i.e., bromoxynil or ioxynil) herbicides as a result of conventional methods of breeding or genetic engineering,
  • auxin hydroxyphen
  • auxinic herbicide-tolerant plants may be tolerant to
  • Such tolerance traits may be expressed, e.g., as 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 cytochrome P450 (CYP450) proteins having an herbicide-degrading activity.
  • AAD or DHT transgenic aryloxyalkanoate dioxygenase
  • BXN 2,2-dich
  • auxinic herbicide-tolerant plants e.g., oilseed rape (canola)
  • auxinic herbicide-tolerant plants are also covered which are, by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such characteristic, rendered able to synthesize one or more insecticidal proteins, especially those known from the bacterial genus Bacillus, particularly from Bacillus thuringiensis, such as ⁇ -endotoxins, e.g., CrylA(b), CrylA(c), CrylF, CryIF(a2), CryIIA(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.
  • VIP1, VIP2, VIP3 or VIP3A insecticidal proteins of bacteria colonizing nematodes
  • 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
  • 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.
  • 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).
  • 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 Diabro
  • Isoptera including species from the families Hodotermitidae, Kalotermitidae, Mastotermitidae, Rhinotermitidae, Serritermitidae, Termitidae, Termopsidae; the tarnished plant bug Lygus lineolaris; 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 psy
  • expression of one or more protein toxins (e.g., insecticidal proteins) in the auxinic-herbicide 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.
  • expression of one or more protein toxins (e.g., insecticidal proteins) in the auxinic-herbicide tolerant plants is effective for controlling cabbage seedpod weevil, the Bertha armyworm, Lygus bugs, or the diamondback moth.
  • auxinic herbicide -tolerant plants e.g., oilseed rape (canola)
  • oilseed rape canola
  • auxinic herbicide -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 trait, 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.
  • auxinic herbicide-tolerant plants e.g., oilseed rape (canola)
  • oilseed rape canola
  • auxinic herbicide-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 trait, 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.
  • auxinic herbicide-tolerant plants e.g., oilseed rape (canola)
  • oilseed rape canola
  • auxinic herbicide-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 trait, 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 ® rape, Dow Agro Sciences, Canada).
  • auxinic herbicide -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 trait, altered to contain increased amounts of vitamins and/or minerals, and/or improved profiles of nutraceutical compounds.
  • auxinic herbicide-tolerant plants described herein, 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-methylsulfmylbutyl- glucosinolate), sulforaphane, 3-indolylmethyl-glucosinolate (glucobrassicin), l-methoxy-3- indolylmethyl-glucosinolate (neoglucobrassicin)); phenolics (e.g., flavonoids (e.g., quercetin, kaempferol), hydroxycinnamoyl derivatives (e.g., l,2,2'-trisinapoylgentiobiose, 1,2- diferuloylgentiobiose, 1 ,2'-disinapoyl-2-feruloylgenti
  • auxinic herbicide-tolerant plants described herein, 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, iso flavones e.g., genistein, daid
  • auxinic herbicide-tolerant plants described herein, 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, taxagifme, 8-benxoyloxy taxagifme, 9-acetyloxy taxusin, 9-hydroxy taxusin, taiwanxam, taxane la, taxane lb, taxane Ic, taxane Id, GMP paclitaxel, 9
  • auxinic herbicide-tolerant plants described herein also can be tolerant to herbicides that inhibit acetohydroxyacid synthase (AHAS).
  • AHAS acetohydroxyacid synthase
  • herein herbicide tolerant AHASL refers to the AHAS large subunit polypeptide expressed from one mutant AHASL allele of an AHASL gene in a plant cell and/or from either or both of two homologous alleles of the same mutant AHASL gene, i.e., in the same genome of, the plant cell, whereby that mutant AHASL can provide herbicide tolerance to an AHAS enzyme of the plant cell.
  • a mutant AHASL gene can be recombinant, or can be obtained by application of a mutagenesis process, a breeding process, or other process known in the art. Such a gene can be hemizygous, heterozygous, or homozygous.
  • A indicates alanine
  • P indicates proline
  • W indicates tryptophan
  • X indicates any amino acid
  • Mutations as compared to the wild-type sequence will be indicated by specifying the wild-type amino acid and position followed by the amino acid present in the mutant.
  • P197X will be used to indicate that the proline at position 197 can be substituted with any amino acid.
  • amino acid numbering system used herein may be the industry standard numbering used for the Arabidopsis thaliana ⁇ At) AHASL sequence, and can be denoted with an ⁇ At).
  • P197( ⁇ 4t) can refer to the proline residue at the position in a Brassica AHASL that corresponds to the proline at position 197 of the Arabidopsis thaliana AHASL.
  • an AHAS herbicide-tolerance-inducing mutation is an alteration in the amino acid sequence of an AHASL enzyme that confers tolerance to one or more herbicides (i.e., sulfonylurea herbicides, imidazolinone herbicides, etc.).
  • herbicides i.e., sulfonylurea herbicides, imidazolinone herbicides, etc.
  • Table 1 provides a non-limiting list of possible sites for AHASL mutations, permissible substitutions, preferred substitutions, and more preferred substitutions.
  • X indicates any amino acid.
  • AHASL mutations can be selected from the group consisting of A122X, P197X, R199X, A205X, S653X, and G654X, and combinations thereof.
  • AHASL mutations can be selected from the group consisting of A122T, A122V, A122D, A122P, A122Y, P197S, P197L, P197T, R199A, R199E, A205V, A205C, A205D, A205E, A205R, A205T, A205W, A205Y, A205N, S653N, S653I, S653F, S653T, G654Q, G654C, W574L, W574M, W574C, W574S, W574R, W574G, W574A, W574F, W574Q, W574Y, G654E, G654D, and combinations thereof.
  • AHASL mutations can be selected from the group consisting of A122T, A122V, R199A, R199E, A205V, S653N, G654E, and combinations thereof.
  • Sources of useful plastidic AHASL genes can be provided from any of the following deposited cell lines listed in Table 2, of Brassica napus (Bn) and Brassica juncea (Bj), wherein their AHAS-inhibitor-tolerant AHAS large subunit (AHASL) alleles are referred to as shown below, with the final letter indicating the Brassica genome (A, B, or C) to which the allele is native: BnAHASLlA or BnAHASLIC for B.
  • the auxinic herbicide -tolerant plants hereof can be inbred varieties, e.g., open-pollinated varieties, or hybrids, e.g., Fl hybrids.
  • the auxinic herbicide-tolerant plants further comprise one or more mutant AHASL genes that may be transgenic or non-transgenic mutant AHAS genes.
  • the AHAS trait can be non-transgenic, i.e., obtained by a process, excluding recombinant DNA techniques, and comprising mutagenesis, genoplasty, and/or isolation of spontaneous mutant plants.
  • Many mutagenesis techniques are known in the art and these can involve application of a mutagenic chemical agent or radiation to seeds, plants parts, or cultured plant cells; alternatively, or in addition, the culturing of plant cells, or the conditions under which plant cells are cultured, can increase the rate of occurrence or accumulation of spontaneous mutations.
  • Genoplasty techniques can include directed mutation-type strategies, such as methods comprising introduction, into the plant cell nucleus, of oligonucleotides that facilitate mismatch-repair-system-mediated nucleotide substitution.
  • auxinic herbicide-tolerant plants further comprising tolerance to at least one ACCase inhibitor herbicide at levels that would normally inhibit the growth of wild-type plant.
  • the auxinic herbicide-tolerant plant described herein expresses an ACCase in which the amino acid sequence differs from an amino acid sequence of an ACCase of a wild-type plant.
  • the amino acid numbering system used herein may be the industry standard numbering system used for the ACCase from Alopecurus myosuroides [Huds.] (also referred to as black grass).
  • the mR A (cDNA) sequence encoding the A. myosuroides ACCase is available at GenBank accession number AJ310767 and the protein sequence is available at GenBank accession No. CAC84161 both of which are specifically incorporated herein by reference.
  • the number of the amino acid referred to will be followed with (Am) to indicate the amino acid in the Alopecurus myosuroides sequence to which the amino acid corresponds.
  • amino acid positions at which an ACCase of an auxininc herbicide -tolerant plant differs from the ACCase of the corresponding wild-type plant include, but are not limited to, one or more of the following positions: ⁇ ,l ⁇ (Am), ⁇ ,l 5(Am), ⁇ ,l%6(Am), ⁇ , ⁇ ⁇ (Am), ⁇ , 24(Am), ⁇ $64(Am), 1,999 (Am), Ifill(Am), 2,039(Am), 2,041 (Am), 2,049(Am), 2,059(Am), 2,014(Am), 2,015(Am), 2,07S(Am), 2,019(Am), 2,080(Am), 2,0Sl (Am), 2,0SS(Am), 2,095(Am), 2,096(Am), or 2,098(Am).
  • a method for treating a plant described herein comprises contacting the plant with an agronomically acceptable composition.
  • the agronomically acceptable composition comprises an auxinic herbicide A.I.
  • Another aspect described herein is a method for preparing a descendent seed.
  • the method comprises planting a seed of or capable of producing a plant described herein.
  • one embodiment described herein is a method for preparing a descendent seed, the method comprising planting a seed of or capable of producing an auxinic herbicide tolerant plant or plant part thereof having the auxinic herbicide-tolerance characteristic of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R, with the proviso that the plant is a monocot or dicot species other than Sinapis arvensis.
  • the method further comprises growing a descendent plant from the seed and harvesting a descendant seed from the descendent plant.
  • the method further comprises applying an auxinic herbicidal composition to the descendent plant.
  • a further aspect described herein is a method for producing a plant product.
  • the method comprises processing a plant or plant part thereof described herein.
  • one embodiment described herein is a method for producing a plant product, the method comprising processing an auxinic herbicide tolerant plant or plant part thereof having the auxinic herbicide-tolerance characteristic of any of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R, with the proviso that the plant is a monocot or dicot species other than Sinapis arvensis, to obtain the plant product therefrom.
  • the plant product is fodder, seed meal, oil, or seed-treatment-coated seeds.
  • the plant part is a seed.
  • herbicide compositions described herein comprise one or more auxinic herbicides that can be chosen from among the compounds listed in Table 4 and their agronomically acceptable salts and esters.
  • auxinic herbicides include these compounds and such salts, esters, or derivatives thereof.
  • auxinic herbicide A.I.s useful herein are shown below in Table 5; these are also applicable to the salt or ester forms thereof.
  • auxinic herbicide tolerant plants described herein comprise tolerance to application of auxinic herbicide in an amount of about 5 to about 5000 g/ha A.I., illustratively, about 10 to about 5000, about 50 to about 4950, about 100 to about 4900, about 150 to about 4850, about 200 to about 4800, about 250 to about 4750, about 300 to about 4700, about 350 to about 4650, about 400 to about 4600, about 450 to about 4550, about 500 to about 4500, about 550 to about 4450, about 600 to about 4400, about 650 to about 3500, about 700 to about 3450, about 750 to about 3400, about 800 to about 3350, about 850 to about 3300, about 900 to about 3250, about 950 to about 3200, about 1000 to about 3150, about 1050 to about 3100, about 1 100 to about 3050, about 1 150 to about 3000, about 1200 to about 2950, about 1250 to about 2900, about 1300 to about 2850, about 13
  • the method can utilize 1 x auxinic herbicide application rates with no significant injury to the plant; in some embodiments thereof, the application rate can exceed 1 x auxinic herbicide; in some embodiments, the rate can be up to 4 x auxinic herbicide, though more typically it will be about 2.5 x or less, or about 2x or less.
  • the herbicide application rate will preferably provide a summed rate that falls within at least about 0.25 x , illustratively, about 0.25 x to about 4x, about 0.5 X to about 3 X , about 1 * to about 2x auxinic herbicide range.
  • the auxinic herbicide composition comprises at least one of clomeprop; cloprop ("3-CPA”); 4-chlorophenoxyacetic acid ("4-CPA”); 2-(4- chlorophenoxy)propionic acid (“4-CPP”); 2,4-dichlorophenoxy acetic acid ("2,4-D”); (3,4- dichlorophenoxy)acetic acid ("3,4-DA”); 4-(2,4-dichlorophenoxy)butyric acid (“2,4-DB”); 2- (3,4-dichlorophenoxy)propionic acid (“3,4-DP”); tris[2-(2,4-dichlorophenoxy)ethyl]phosphite (“2,4-DEP”); dichlorprop ("2,4-DP”); 2,4,5-trichlorophenoxyacetic acid ("2,4,5-T”); fenoprop ("2,4,5-TP”); 2-(4-chloro-2-methylphenoxy)acetic acid (“MCPA”); 4-(4-chloro
  • the auxinic herbicide composition comprises at least one of clomeprop; cloprop ("3-CPA”); 4-chlorophenoxyacetic acid ("4-CPA”); 2-(4- chlorophenoxy)propionic acid (“4-CPP”); (3,4-dichlorophenoxy)acetic acid ("3,4-DA”); 4-(2,4- dichlorophenoxy)butyric acid ("2,4-DB”); 2-(3,4-dichlorophenoxy)propionic acid (“3,4-DP”); tris[2-(2,4-dichlorophenoxy)ethyl]phosphite (“2,4-DEP”); dichlorprop ("2,4-DP”); 2,4,5- trichlorophenoxyacetic acid ("2,4,5-T”); fenoprop ("2,4,5-TP”); 4-(4-chloro-2- methylphenoxy)butyric acid (“MCPB”); chloramben; dicamba; tricamba; 2,3,6-trichlorobenzoic
  • the auxinic herbicide composition comprises at least one of: 4- (2,4-dichlorophenoxy)butyric acid ("2,4-DB”); dicamba; aminopyralid; picloram; or quinclorac.
  • the auxinic herbicide composition comprises at least one of: aminopyralid or picloram.
  • the auxinic herbicide composition comprises dicamba.
  • the herbicide compositions described herein can further comprise one or more agronomically acceptable A.I.(s), e.g., herbicidal A.I.s.
  • agronomically acceptable A.I.(s) include the A.I.s and their agronomically acceptable salts and esters.
  • herbicides include, but are not limited to, AHAS inhibitors; bleaching herbicides such as hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors or phytoene desaturase (PDS) inhibitors; enolpyruvyl shikimate 3-phosphate synthase (EPSPS) inhibitors such as glyphosate; glutamine synthetase (GS) inhibitors such as glufosinate; protoporphyrinogen-IX oxidase (PPO) inhibitors; lipid biosynthesis inhibitors such as ACCase inhibitors; or oxynil (i.e., bromoxynil or ioxynil) herbicides.
  • HPPD hydroxyphenylpyruvate dioxygenase
  • PDS phytoene desaturase
  • EPSPS enolpyruvyl shikimate 3-phosphate synthase
  • GS glutamine synthetase
  • PPO protoporphy
  • AHAS -inhibitor herbicides include, e.g., imidazolinone herbicides, one or more SU herbicides selected from the group consisting of amidosulfuron, flupyrsulfuron, foramsulfuron, imazosulfuron, iodosulfuron, mesosulfuron, nicosulfuron, thifensulfuron, and tribenuron, agronomically acceptable salts and esters thereof, and combinations thereof.
  • the herbicide compositions described herein can further comprise one or more agronomically acceptable A.I.(s) of other classes, e.g., agronomic fungicides, bactericides, algicides, nematicides, insecticides, and the like (e.g., malathion, pyrethrins/pyrethrum, carbaryl, spinosad, permethrin, bifenthrin, and esfenvalerate).
  • A.I.(s) of other classes e.g., agronomic fungicides, bactericides, algicides, nematicides, insecticides, and the like (e.g., malathion, pyrethrins/pyrethrum, carbaryl, spinosad, permethrin, bifenthrin, and esfenvalerate).
  • the herbicidal compositions hereof comprising one or more auxinic herbicides, and optionally other agronomic A.I.(s) can be used in any agronomically acceptable format.
  • these can be formulated as ready-to-spray aqueous solutions, powders, suspensions; as concentrated or highly concentrated aqueous, oily, or other solutions, suspensions, or dispersions; as emulsions, oil dispersions, pastes, dusts, granules, or other broadcastable formats.
  • the herbicide compositions can be applied by any means known in the art, including, for example, spraying, atomizing, dusting, spreading, watering, seed treatment, or co-planting in admixture with the seed.
  • an herbicidal composition hereof can comprise, e.g., a combination of: auxinic herbicide(s), e.g., dicamba; AHAS-inhibitor(s), e.g., imidazolinone(s) and/or sulfonylurea(s); ACCase-inhibitor(s); EPSPS inhibitor(s), e.g., glyphosate; glutamine synthetase inhibitor(s), e.g.., glufosinate; protoporphyrinogen-IX oxidase (PPO) inhibitor(s), e.g., saflufenacil; fungicide(s), e.g., strobilurin fungicide(s) such as pyraclostrobin; and the like.
  • auxinic herbicide(s) e.g., dicamba
  • AHAS-inhibitor(s) e.g., imida
  • an herbicidal composition hereof can comprise, e.g., a combination of auxinic herbicide(s), e.g., dicamba; and strobilurin fungicide(s) such as pyraclostrobin(s).
  • auxinic herbicide(s) e.g., dicamba
  • strobilurin fungicide(s) such as pyraclostrobin(s).
  • An herbicidal composition will be selected according to the tolerances of a plant hereof, and the plant can be selected from among those having stacked tolerance traits.
  • the plant to be used is selected from among those that further comprise a trait of tolerance to such herbicide.
  • Such further tolerance traits can be provided to the plant by any method known in the art, e.g., including techniques of traditional breeding to obtain a tolerance trait gene by hybridization or introgression, of mutagenesis, and/or of transformation. Such plants can be described as having "stacked" traits.
  • Optional A.I.s of other types include, but are not limited to fungicides such as strobilurins, e.g., pyraclostrobin, alone or in combination with, e.g., boscalid, epiconazole, metaconazole, tebuconazole, kresoxim-methyl, fluxapyroxad, isopyrazam, flutolanil, and the like; insecticides such as lepidoptericides, coleoptericides, and the like; nematicides; molluskicides; and others known in the art.
  • fungicides such as strobilurins, e.g., pyraclostrobin, alone or in combination with, e.g., boscalid, epiconazole, metaconazole, tebuconazole, kresoxim-methyl, fluxapyroxad, isopyrazam, flutolanil, and the like
  • insecticides such as lepid
  • optional A.I.s include fungicides such as strobilurins (e.g., pyraclostrobin), boscalid, epiconazole, metaconazole, tebuconazole, kresoxim-methyl, fluxapyroxad, isopyrazam, flutolanil, and the like, each alone or in combination.
  • strobilurins e.g., pyraclostrobin
  • boscalid e.g., pyraclostrobin
  • epiconazole e.g., metaconazole, tebuconazole, kresoxim-methyl, fluxapyroxad, isopyrazam, flutolanil, and the like, each alone or in combination.
  • suitable examples of herbicides that are ACCase inhibitors include, but are not limited to, cyclohexanedione herbicides (DIMs, also referred to as: cyclohexene oxime cyclohexanedione oxime; and CHD), aryloxyphenoxy propionate herbicides (also referred to as aryloxyphenoxy propanoate; aryloxyphenoxyalkanoate; oxyphenoxy; APP; AOPP; APA; APPA; FOP, note that these are sometime written with the suffix '-oic'), and phenylpyrazole herbicides (also known as DENs; and sometimes referred to under the more general class of Phenylpyrazole such as pinoxaden (e.g., herbicides sold under the trade names Axial and Traxos).
  • DIMs cyclohexanedione herbicides
  • aryloxyphenoxy propionate herbicides also
  • At least one herbicide is selected from the group consisting of sethoxydim, cycloxydim, tepraloxydim, haloxyfop, haloxyfop-P or a derivative of any of these herbicides.
  • Table 6 lists herbicides that interfere with ACCase activity.
  • the herbicidal compositions comprising an auxinic herbicide, and optionally other agronomic A.I.(s) and/or their agriculturally suitable salts and esters can also comprise auxiliaries, which are customary for the formulation of crop protection agents.
  • auxiliaries customary for the formulation of crop protection agents include inert auxiliaries, solid carriers, surfactants (such as dispersants, protective colloids, emulsifiers, wetting agents and tackifiers), organic and inorganic thickeners, penetrants (such as penentration-enhancing organosilicone surfactants or acidic sulfate chelates, e.g., CT-301TM available from Cheltec, Inc.), safeners, bactericides, antifreeze agents, antifoams, colorants, and adhesives.
  • Formulations of the herbicide compositions useful herein can be prepared according to any method known useful therefor in the art.
  • thickeners i.e., compounds which impart to the formulation modified flow properties, i.e., high viscosity in the state of rest and low viscosity in motion
  • thickeners are polysaccharides, such as xanthan gum (Kelzan ® from Kelco), Rhodopol ® 23 (Rhone Poulenc) or Veegum ® (from R.T. Vanderbilt), and also organic and inorganic sheet minerals, such as Attaclay ® (from Engelhardt).
  • antifoams examples include silicone emulsions (such as, for example, Silikon ® SRE,
  • Wacker or Rhodorsil ® from Rhodia long-chain alcohols, fatty acids, salts of fatty acids, organofluorine compounds and mixtures thereof.
  • antifreeze agents are ethylene glycol, propylene glycol, urea, or glycerol.
  • colorants include members of colorant classes such as the sparingly water- soluble pigments and the water-soluble dyes. Some specific examples of these include the dyes known under the names Rhodamin B, C.I. Pigment Red 112 and C.I.
  • Solvent Red 1 and also 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 112, 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.
  • adhesives examples include polyvinylpyrrolidone, polyvinyl acetate, polyvinyl alcohol, and tylose.
  • Suitable inert auxiliaries are, for example, the following: mineral oil fractions of medium to high boiling point, such as kerosene and diesel oil, furthermore coal tar oils and oils of vegetable or animal origin, aliphatic, cyclic and aromatic hydrocarbons, for example paraffin, tetrahydronaphthalene, alkylated naphthalenes and their derivatives, alkylated benzenes and their derivatives, alcohols such as methanol, ethanol, propanol, butanol and cyclohexanol, ketones such as cyclohexanone or strongly polar solvents, for example amines such as N- methylpyrrolidone, and water.
  • mineral oil fractions of medium to high boiling point such as kerosene and diesel oil, furthermore coal tar oils and oils of vegetable or animal origin
  • aliphatic, cyclic and aromatic hydrocarbons for example paraffin, tetrahydronaphthalene, alkylated na
  • Suitable carriers include liquid and solid carriers.
  • Solid carriers include e.g., mineral earths such as silicas, silica gels, silicates, talc, kaolin, limestone, lime, chalk, bole, loess, clay, dolomite, diatomaceous earth, calcium sulfate, magnesium sulfate and magnesium oxide, ground synthetic materials, fertilizers such as ammonium sulfate, ammonium phosphate, ammonium nitrate and ureas, and products of vegetable origin, such as cereal meal, tree bark meal, wood meal and nutshell meal, cellulose powders, or other solid carriers.
  • mineral earths such as silicas, silica gels, silicates, talc, kaolin, limestone, lime, chalk, bole, loess, clay, dolomite, diatomaceous earth, calcium sulfate, magnesium sulfate and magnesium oxide, ground synthetic materials, fertilizers such as ammonium sulfate, ammonium phosphate, ammonium n
  • Suitable surfactants are the alkali metal salts, alkaline earth metal salts and ammonium salts of aromatic sulfonic acids, for example lignosulfonic acids (e.g., Borrespers-types, Borregaard), phenolsulfonic acids, naphthalenesulfonic acids (Morwet types, Akzo Nobel) and dibutylnaphthalenesulfonic acid (Nekal types, BASF AG), and of fatty acids, alkyl- and alkylarylsulfonates, alkyl sulfates, lauryl ether sulfates and fatty alcohol sulfates, and salts of sulfated hexa-, hepta- and octadecanols, and also of fatty alcohol glycol ethers, condensates of sulfonated
  • aromatic sulfonic acids for example lignosulfonic acids (e.g., Borrespers
  • Granules for example, coated granules, impregnated granules and homogeneous granules, can be prepared by binding the A.I.s to solid carriers.
  • Aqueous use forms can be prepared from emulsion concentrates, suspensions, pastes, wettable powders, or water-dispersible granules by adding water.
  • the herbicidal compositions comprising an auxinic herbicide, and optionally other agronomic A.I.(s) and/or their agriculturally suitable salts and esters, either as such or dissolved in an oil or solvent, can be homogenized in water by means of a wetting agent, tackifier, dispersant or emulsifier.
  • a wetting agent e.g., tackifier, dispersant or emulsifier
  • concentrates comprising active compound, wetting agent, tackifier, dispersant or emulsifier and, if desired, solvent or oil, which are suitable for dilution with water.
  • the concentrations of the herbicides present in the herbicidal composition comprising an auxinic herbicide, and optionally other agronomic A.I.(s) and/or their agriculturally suitable salts and esters can be varied within wide ranges.
  • the formulations comprise approximately from 0.001 to 98% by weight, preferably 0.01 to 95% by weight of at least one active ingredient.
  • the A.I.s are employed in a purity of from 90% to 100%, preferably 95% to 100%) (according to NMR spectrum).
  • the herbicides are present in suspended, emulsified, or dissolved form.
  • the formulation described herein can be in the form of aqueous solutions, powders, suspensions, also highly-concentrated aqueous, oily, or other suspensions or dispersions, aqueous emulsions, aqueous microemulsions, aqueous suspo-emulsions, oil dispersions, pastes, dusts, materials for spreading or granules.
  • the herbicides can, for example, be formulated as follows:
  • active compound 10 parts by weight of active compound are dissolved in 90 parts by weight of water or a water-soluble solvent.
  • wetting agent(s) or other adjuvants are added.
  • the active compound dissolves upon dilution with water. This gives a formulation with an active compound content of 10% by weight.
  • active compound 20 parts by weight of active compound are dissolved in 70 parts by weight of cyclohexanone with addition of 10 parts by weight of a dispersant, for example polyvinylpyrrolidone. Dilution with water gives a dispersion.
  • a dispersant for example polyvinylpyrrolidone.
  • the active compound content is 20% by weight.
  • active compound 15 parts by weight of active compound are dissolved in 75 parts by weight of an organic solvent (e.g., alkylaromatics) with addition of calcium dodecylbenzenesulfonate and castor oil ethoxylate (in each case 5 parts by weight). Dilution with water gives an emulsion.
  • the formulation has an active compound content of 15% by weight.
  • Water-dispersible granules and water-soluble granules 50 parts by weight of active compound are ground finely with addition of 50 parts by weight of dispersants and wetting agent(s) and made into water-dispersible or water-soluble granules by means of technical appliances (for example extrusion, spray tower, fluidized bed). Dilution with water gives a stable dispersion or solution of the active compound.
  • the formulation has an active compound content of 50% by weight.
  • active compound 0.5 parts by weight are ground finely and associated with 99.5 parts by weight of carriers. Current methods here are extrusion, spray-drying, or the fluidized bed. This gives granules to be applied undiluted with an active compound content of 0.5%> by weight.
  • Aqueous use forms can be prepared from emulsion concentrates, suspensions, pastes, wettable powders, or water-dispersible granules by adding water.
  • the herbicides or the herbicidal compositions comprising them can be applied pre-, post- emergence or pre -plant, or together with the seed. It is also possible to apply the herbicidal composition or active compounds by applying seed, pretreated with the herbicidal compositions or active compounds, of a crop plant.
  • the herbicides or herbicidal compositions can be applied by treating seed.
  • the treatment of seeds comprises essentially all procedures familiar to the person skilled in the art (e.g., seed dressing, seed coating, seed dusting, seed soaking, seed film coating, seed multilayer coating, seed encrusting, seed dripping and seed pelleting).
  • the herbicidal compositions can be applied diluted or undiluted.
  • One embodiment is a method for treating a seed comprising: (a) providing a seed comprising nucleic acid of any one of SEQ ID NOs: 8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, the expression of the nucleic acid conferring to the plant or seed tolerance to an auxinic herbicide; and (b) contacting said seed with an agronomically acceptable composition.
  • the auxinic herbicides, and optionally other agronomic A.I.(s) can be mixed with a large number of representatives of other herbicidal or growth-regulating active ingredient groups and then applied concomitantly.
  • Suitable components for mixtures are, for example, 1,2,4-thiadiazoles, 1,3,4-thiadiazoles, amides, aminophosphoric acid and its derivatives, aminotriazoles, anilides, (het)aryloxyalkanoic acids and their derivatives, benzoic acid and its derivatives, benzothiadiazinones, 2-aroyl-l,3-cyclohexanediones, 2-hetaroyl-l,3- cyclohexane-diones, hetaryl aryl ketones, benzylisoxazolidinones, meta-CF3 -phenyl derivatives, carbamates, quinolinecarboxylic acid and its derivatives, chloroacetanilides, cyclo
  • auxinic herbicide members are excluded from the following classes: benzoic acid and its derivatives; quinolinecarboxylic acid and its derivatives; 2- phenylpropionic acid and its derivatives; and pyridinecarboxylic acid and its derivatives.
  • herbicides alone or in combination with other herbicides, or else in the form of a mixture with other crop protection agents, for example together with agents for controlling pests or phytopathogenic fungi or bacteria.
  • miscibility with mineral salt solutions which are employed for treating nutritional and trace element deficiencies.
  • additives such as non-phytotoxic oils and oil concentrates can also be added.
  • Safeners are chemical compounds, which prevent or reduce herbicide-induced injury to useful plants without having a major impact on the herbicidal action of the herbicides. 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. The safeners and the herbicides can be applied simultaneously or in succession.
  • Suitable safeners are e.g., (quinolin-8-oxy)acetic acids, l-phenyl-5-haloalkyl-lH-l,2,4- triazol-3-carboxylic acids, l-phenyl-4,5-dihydro-5-alkyl-lH-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-phenyl-
  • safeners are benoxacor, cloquintocet, cyometrinil, cyprosulfamide, dichlormid, dicyclonon, dietholate, fenchlorazole, fenclorim, flurazole, fluxofenim, furilazole, isoxadifen, mefenpyr, mephenate, naphthalic anhydride, oxabetrinil, 4-(dichloroacetyl)-l-oxa-4- azaspiro[4.5]decane (MON4660, CAS 71526-07-3) and 2,2,5-trimethyl-3-(dichloroacetyl)-l,3- oxazolidine (R-29148, CAS 52836-31-4).
  • herbicides and/or safeners 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 described herein. Furthermore, some of the above mentioned herbicides and/or safeners 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 described herein.
  • aryloxyphenoxy propionate herbicides are chiral, and some of them are commonly used in enantiomerically enriched or enantiopure form, e.g., clodinafop, cyhalofop, fenoxaprop-P, fluazifop-P, haloxyfop-P, metamifop, propaquizafop or quizalofop-P.
  • glufosinate may be used in enantiomerically enriched or enantiopure form, also known as glufosinate-P.
  • Herbicide -tolerant plants described herein can be used in conjunction with an herbicide to which they are tolerant.
  • Herbicides can be applied to the plants described herein 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.
  • One embodiment described herein is a method for controlling weeds at a locus for growth of a plant or plant part thereof, the method comprising: applying a composition comprising an auxinic herbicide to the locus.
  • the locus for growth of a plant is a field.
  • the plant has the auxinic herbicide-tolerance characteristic of any of Sinapis arvensis lines DT-01 SA2-R or DT-01 BC8SA2-R.
  • the plant has the auxinic herbicide- tolerance characteristic DART.
  • One embodiment described herein is a method for controlling weeds in a field by an application of an auxinic herbicide without significantly inhibiting the growth of a Brassica plant, the method comprising: (a) providing a Brassica plant or seed comprising the nucleic acid of: (i) a chimeric polynucleotide comprising both a Sinapis arvensis polynucleotide portion and a Brassica polynucleotide portion, wherein said chimeric polynucleotide encodes the DART trait of any one of lines DT-01 Cyc2 BNS4, DT-01 BC3Bn#13, DT-01 BC4Bn#13-l, DT-01 BC5Bn#13-l-18, DT-01 SA2-R, or DT-01 BC8SA2-R, a representative sample of seed of each line having been deposited with American Type Culture Collection (ATCC) under Patent Deposit Designation Numbers PTA-120132, PTA-11211, PTA-12050, PTA-11212, PTA-1121
  • Another embodiment described herein is a method for controlling weeds in a field by application of an auxinic herbicide without significantly inhibiting the growth of a Brassica plant, the method comprising: (a) providing a Brassica plant or seed comprising the nucleic acid of any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, the expression of the nucleic acid conferring to the plant or seed tolerance to an auxinic herbicide; and (b) applying an herbicide composition comprising an effective amount of an auxinic herbicide: (i) to the field, followed by planting of said plant or seed in therein; (ii) to the field, during or after planting of said seed therein; (iii) to the plant in said field and to weeds in the vicinity of the plant; (iv) to said seed, followed by planting of said seed in the field; or (v) to a plant by the seed after it has been planted in the field, and to weed
  • Another embodiment described herein is a method for combating undesired vegetation in a field comprising contacting a seed of a crop plant before sowing and/or after pregermination with an auxinic herbicide composition without significantly inhibiting the growth of said crop plant.
  • the seed comprises the DART auxinic herbicide-tolerance trait.
  • the auxinic herbicide composition is applied to the weeds and to the plant, seed, or the plant produced by the seed.
  • Herbicide compositions described herein 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.
  • herbicides can be used to control the growth of weeds that may be found growing in the vicinity of the herbicide -tolerant plants described herein.
  • an herbicide can be applied to a plot in which herbicide-tolerant plants described herein are growing in vicinity to weeds.
  • An herbicide to which the herbicide-tolerant plant described herein 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.
  • Anther embodiment described herein is a method for controlling weeds in the vicinity of an auxinic herbicide -tolerant plant as described herein.
  • the method comprises applying an effective amount of an auxinic herbicide 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.
  • the auxinic herbicide-tolerant plants described herein 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.
  • Another embodiment described herein is a method for controlling weeds in a field or crops by use of an auxinic herbicide without significantly inhibiting the growth of the crop plant, the method comprising: (a) providing a seed-treatment-treated seed comprising the nucleic acid of any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, the expression of the nucleic acid conferring to the plant or seed tolerance to an auxinic herbicide, the seed treatment comprising an auxinic herbicide; and (b) planting said treated seed in the field.
  • herbicide(s) e.g., auxinic herbicides
  • 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.
  • Such compositions are described herein as seed treatment compositions.
  • the seed treatment composition comprises an auxinic herbicide.
  • the seed has, disposed on a surface thereof, a seed treatment composition comprising at least one agronomically acceptable ingredient.
  • the ingredient is at least one agronomically acceptable herbicide, fungicide, nematicide, or insecticide, or a combination thereof.
  • the said insecticide comprises at least one anti-coleopteran agent, anti-hemipteran agent, anti-lepidopteran agent, or a combination thereof.
  • Another embodiment described herein is a method for treating the seed with an agronomically acceptable composition, the method comprising contacting the seed with the agronomically acceptable composition, wherein the composition comprises an auxinic herbicide.
  • Another embodiment described herein is a method for preparing a treated seed, the method comprising: providing a seed and applying thereto a seed treatment composition.
  • 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.
  • suitable binders are block copolymers EO/PO surfactants but also polyvinylalcohols, polyvinylpyrrolidones, polyacrylates, polymethacrylates, polybutenes, polyisobutylenes, polystyrene, polyethyleneamines, polyethyleneamides, polyethyleneimines (Lupasol ® , Polymin ® ), polyethers, polyurethanes, polyvinylacetate, tylose and copolymers derived from these polymers.
  • colorants can be included in the formulation.
  • Suitable colorants or dyes for seed treatment formulations are Rhodamin B, C.I. Pigment Red 112, 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 112, 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.
  • 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.
  • One embodiment described herein is a method of treating soil by the application, in particular into the seed drill: either of a granular formulation containing the auxinic herbicide 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.
  • Another embodiment described herein comprises seeds coated with and/or containing a seed treatment formulation comprising auxinic herbicide and at least one other herbicide such as, e.g., an AH AS -inhibitor 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, triasul
  • 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.
  • the seed treatment application with auxinic herbicide or with a formulation comprising the auxinic herbicide is carried out by spraying or dusting the seeds before sowing and before emergence of the plants.
  • the corresponding formulations are applied by treating the seeds with an effective amount of auxinic herbicide or a formulation comprising the auxinic herbicide.
  • Another aspect described herein is a method for combating undesired vegetation or controlling weeds comprising: contacting the seeds of the auxinic herbicide-tolerant plants described herein before sowing and/or after pregermination with auxinic herbicide.
  • the method can further comprise sowing the seeds, for example, in soil, in a field, or in a potting medium in a 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 that grow in locations where they are undesired.
  • the weeds described herein 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, Centaur ea, 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.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame i3 ⁇ 4r_R_8134.g27742.tl (SEQ ID NO:8), and encodes a translation elongation factor EF1 A/initiation factor family protein (SEQ ID NO: 9) to provide the auxin herbicide tolerance. See Table 7.
  • the nucleic acid of SEQ ID NO: 8 that encodes the polypeptide of SEQ ID NO: 9 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises SEQ ID NO: 10 and encodes a translation elongation factor EF1 A/initiation factor family protein (SEQ ID NO: 11).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_2008.g8030.tl (SEQ ID NO: 12), and encodes a KAT2 peroxisomal 3- ketoacyl-CoA thiolase 3 (SEQ ID NO: 13) to provide the auxin herbicide tolerance. See Table 8.
  • the nucleic acid of SEQ ID NO: 12 that encodes the polypeptide of SEQ ID NO: 13 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises Sar_S_22769.g45642.tl (SEQ ID NO: 14), and encodes a KAT2 peroxisomal 3-ketoacyl-CoA thiolase 3 (SEQ ID NO: 15).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide. Table 8. S. arvensis Auxin-Herbicide Tolerance Trait Sequence Gene 2
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame is 5ar_R_942.g3734.tl (SEQ ID NO: 16), and encodes a calmodulin 5-like protein that facilitates protein-protein interactions (SEQ ID NO: 17) to provide the auxin herbicide tolerance. See Table 9. No auxin herbicide susceptible counterpart was identified.
  • the nucleic acid of SEQ ID NO: 16 that encodes the polypeptide of SEQ ID NO: 17 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_6352.g22471.tl (SEQ ID NO: 18), and encodes an ARF21 transcription factor (SEQ ID NO: 19) to provide the auxin herbicide tolerance.. See Table 10.
  • the nucleic acid of SEQ ID NO: 18 that encodes the polypeptide of SEQ ID NO: 19 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises Sar_S_14769.g33259.tl (SEQ ID NO:20), and encodes an ARF21 transcription factor (SEQ ID NO:21).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • nucleic acid that provides auxin herbicide tolerance to a plant.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame is Sar_R_5279.gl9199.tl (SEQ ID NO:22), and encodes a TPR3 transcription factor (SEQ ID NO:23) to provide the auxin herbicide tolerance. See Table 1 1.
  • the nucleic acid of SEQ ID NO:22 that encodes the polypeptide of SEQ ID NO:23 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises Sar_S_4l 12.gl0923.tl (SEQ ID NO:24), and encodes a TPR3 transcription factor (SEQ ID NO:25).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar R l 119.g4334.tl (SEQ ID NO:26), and encodes a Chloroplast Trans- Membrane, Auxin- Associated Protein- 1 (cpTAAP-1) (SEQ ID NO:27) to provide the auxin herbicide tolerance. See Table 12.
  • the nucleic acid of SEQ ID NO:26 that encodes the polypeptide of SEQ ID NO:27 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises Sar_S_31217.g55588.tl (SEQ ID NO:28), and encodes a Chloroplast Trans-Membrane, Auxin- Associated Protein- 1 (cpTAAP-1) (SEQ ID NO:29).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • Sar_S_31217.g55588.tl Chloroplast Trans-Membrane, Auxin-Associated Protein-1 (cpTAAP-1)
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame 5ar_R_21396.g52442.tl (SEQ ID NO:30), and encodes an Endo-Mitochondrial, Auxin- Associated Protein-1 (mtAAP-1) (SEQ ID NO:31) to provide the auxin herbicide tolerance. See Table 13.
  • the nucleic acid of SEQ ID NO:30 that encodes the polypeptide of SEQ ID NO: 31 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises iSar_S_20600.g42686.tl (SEQ ID NO:32), and encodes an Endo-Mitochondrial, Auxin-Associated Protein-1 (mtAAP-1) (SEQ ID NO:33).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame iSar_R_3394.gl2813.tl (SEQ ID NO:34), and encodes a zinc-binding ribosomal protein family protein (SEQ ID NO:35) to provide the auxin herbicide tolerance. See Table 14.
  • the nucleic acid of SEQ ID NO:34 that encodes the polypeptide of SEQ ID NO:35 provides the DART auxinic herbicide tolerance trait to a plant. No wild type auxin herbicide susceptible counterpart was identified.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_4824.gl7686.tl (SEQ ID NO:36), and encodes a SAUR-like auxin- responsive protein family transcription factor (SEQ ID NO: 37) to provide the auxin herbicide tolerance. See Table 15.
  • the nucleic acid of SEQ ID NO:36 that encodes the polypeptide of SEQ ID NO:37 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises 5ar_S_12132.g28371.tl (SEQ ID NO:38), and encodes a SAUR-like auxin-responsive protein family transcription factor (SEQ ID NO:39).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame S * ar_R_38470.g64923.t (SEQ ID NO:40), and encodes a HTA13, histone protein (SEQ ID NO:41) to provide the auxin herbicide tolerance. See Table 16.
  • the nucleic acid of SEQ ID NO:40 that encodes the polypeptide of SEQ ID NO:41 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises iSar_S_4108.gl0914.tl (SEQ ID NO:42), and encodes a HTA13, histone protein (SEQ ID NO:43).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_706.G2840.Tl (SEQ ID NO:44), and encodes a knotted 1 like (K AT4) transcription factor (SEQ ID NO:45) to provide the auxin herbicide tolerance. See Table 17.
  • the nucleic acid of SEQ ID NO:44 that encodes the polypeptide of SEQ ID NO:45 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises Sar_S_5265.G13674.Tl (SEQ ID NO:46), and encodes a knotted 1 like (K AT4) transcription factor (SEQ ID NO:47).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_19858.g50467.tl (SEQ ID NO:48), and encodes a Zinc-Finger-Like, Auxin- Associated Protein- 1 (zFAAP-1) (SEQ ID NO:49) to provide the auxin herbicide tolerance. See Table 18.
  • the nucleic acid of SEQ ID NO:48 that encodes the polypeptide of SEQ ID NO:49 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises 5ar_S_68453.g74218.tl (SEQ ID NO:50), and encodes a Zinc-Finger-Like, Auxin-Associated Protein-1 (zFAAP-1) (SEQ ID NO:51).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_10390.g33598.tl (SEQ ID NO:52), and encodes an Alba DNA/RNA- binding protein transcription factor (SEQ ID NO:53) to provide the auxin herbicide tolerance. See Table 19.
  • the nucleic acid of SEQ ID NO:52 that encodes the polypeptide of SEQ ID NO:53 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises Sar_S_2341 l .g46486.tl (SEQ ID NO:54), and encodes an Alba DNA/RNA-binding protein transcription factor (SEQ ID NO:55).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame 5ar_R_2411.G9430.Tl (SEQ ID NO:56), and encodes an IAA16 transcription factor (SEQ ID NO:57) to provide the auxin herbicide tolerance. See Table 20.
  • the nucleic acid of SEQ ID NO:56 that encodes the polypeptide of SEQ ID NO:57 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises 5ar_S_29291.G53554.Tl (SEQ ID NO:58), and encodes an IAA16 transcription factor (SEQ ID NO:59).
  • a second auxin herbicide susceptible nucleic acid open reading frame is 5ar_S_3202.G8615.Tl (SEQ ID NO:60), and encodes an IAA16 transcription factor (SEQ ID NO:61).
  • either of the auxin herbicide susceptible nucleic acids is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_21966.G53064.Tl (SEQ ID NO: 62), and encodes an IAA12 bodenlos/monopteros transcription factor (SEQ ID NO:63) to provide the auxin herbicide tolerance. See Table 21.
  • the nucleic acid of SEQ ID NO:62 that encodes the polypeptide of SEQ ID NO: 63 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises &r_S_12886.G29900.Tl/ Sar_S_59713.G71763.Tl (SEQ ID NO:64), and encodes an IAA12 bodenlos/monopteros transcription factor (SEQ ID NO:65).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from 5. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame 5ar_R_36336.g63877.tl (SEQ ID NO:66), and encodes RUBl (SEQ ID NO:67) to provide the auxin herbicide tolerance. See Table 22.
  • the nucleic acid of SEQ ID NO: 66 that encodes the polypeptide of SEQ ID NO: 67 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises 5ar_S_7171.gl8066.tl (SEQ ID NO:68), and encodes RUBl (SEQ ID NO: 69). See Table 22.
  • an additional auxin herbicide susceptible nucleic acid open reading frame is 5ar_S_16564.g36270.tl (SEQ ID NO:70), and encodes RUBl (SEQ ID NO:71).
  • an additional auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises 5ar_S_19347.g40811.tl (SEQ ID NO:72), and encodes RUBl (SEQ ID NO:73).
  • an additional auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises 5ar_S_23559.g46658.tl (SEQ ID NO:74), and encodes RUBl (SEQ ID NO:75). See Table 22.
  • any of the auxin herbicide susceptible nucleic acids is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide. Table 22. S. arvensis Auxin-Herbicide Tolerance Trait Sequence Genes 17-19
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame 5ar_R_16466.g45572.tl (SEQ ID NO:76), and encodes a CAM7 transcription factor (SEQ ID NO:77) to provide the auxin herbicide tolerance. See Table 23.
  • the nucleic acid of SEQ ID NO: 76 that encodes the polypeptide of SEQ ID NO: 77 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises 5ar_S_29545.g53829.tl (SEQ ID NO:78), and encodes a CAM7transcription factor (SEQ ID NO: 79).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the auxinic herbicide tolerance trait is DART.
  • the nucleic acid is an isolated, mutagenized, or recombinant nucleic acid from S. arvensis.
  • the auxin herbicide tolerance nucleic acid comprises the open reading frame Sar_R_3970.gl4672.tl (SEQ ID NO:80), and encodes a cytochrome P450 CYP83A1 (SEQ ID NO:81) to provide the auxin herbicide tolerance. See Table 24.
  • the nucleic acid of SEQ ID NO:80 that encodes the polypeptide of SEQ ID NO:81 provides the DART auxinic herbicide tolerance trait to a plant.
  • the auxin herbicide susceptible nucleic acid corresponding to the auxin herbicide tolerance nucleic acid comprises Sar_S_51371.g68743.tl (SEQ ID NO:82), and encodes a cytochrome P450 CYP83A1 (SEQ ID NO:83).
  • the auxin herbicide susceptible nucleic acid is mutagenized to create an auxin herbicide tolerant polynucleotide encoding an auxin herbicide tolerant polypeptide.
  • the nucleic acids comprise polynucleotides having nucleotide sequences that encode polypeptides comprising the amino acid sequences of any one of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81.
  • the nucleic acid molecules comprise polynucleotides comprising the nucleotide sequences of any one of SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, or degenerate, homologous, or codon-optimized variants thereof.
  • a polynucleotide having a nucleotide sequence at least, for example, 90% "identical" to a reference nucleotide sequence encoding a polypeptide, as used herein, is intended to mean that the nucleotide sequence of the polynucleotide be identical to the reference sequence except that the polynucleotide sequence can include up to about ten substitutions or point mutations per each 100 nucleotides of the reference nucleotide sequence.
  • nucleotide having a nucleotide sequence about 0% identical to a reference nucleotide sequence up to 10% of the nucleotides in the reference sequence can be deleted or substituted with another nucleotide, or a number of nucleotides up to 10%> of the total nucleotides in the reference sequence can be inserted into the reference sequence.
  • substitutions or mutations of the reference sequence can occur at the 5'- or 3 '-terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence, or in one or more contiguous groups within the reference sequence.
  • two or more polynucleotide sequences can be compared by determining their percent identity (or "homology").
  • Two or more amino acid sequences likewise can be compared by determining their percent identity.
  • the percent identity of two sequences, whether nucleic acid or peptide sequences, is generally described as the number of exact matches between two aligned sequences divided by the length of the shorter sequence and multiplied by 100. Numerous applications are available for preforming sequence comparisons and alignments, e.g., BLAST. Altschul et al, (1990); Altschul et al, (1997).
  • nucleic acid molecules having a sequence at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid sequence shown in SEQ ID NOs:8, 12, 16, 18, 22, 26, 30, 34, 36, 40, 44, 48, 52, 56, 62, 66, 76, or 80, or degenerate, homologous, or codon-optimized variants thereof.
  • polypeptides described herein include those encoding mutations, variations, substitutions, and particular examples of the polypeptides described herein.
  • guidance concerning how to make phenotypically silent amino acid substitutions is known in the art.
  • fragments, derivatives, or analogs of the polypeptides of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81 can be (i) ones in which one or more of the amino acid residues (e.g., 1, 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 residues, or even more) are substituted with a conserved or non-conserved amino acid residue (preferably a conserved amino acid residue).
  • Such substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) ones in which one or more of the amino acid residues includes a substituent group (e.g., 1, 2, 3, 4, 5, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50 residues or even more), or (iii) ones in which the mature polypeptide is fused with another compound, such as a compound to increase the half-life of the polypeptide (for example, polyethylene glycol), or (iv) ones in which the additional amino acids are fused to a mature polypeptide, such as an IgG Fc fusion region peptide or leader or secretory sequence or a sequence which is employed for purification of the mature polypeptide or a proprotein sequence.
  • a substituent group e.g., 1, 2, 3, 4, 5, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50 residues or even more
  • a substituent group e.g., 1, 2, 3, 4, 5, 7, 8, 9, 10, 15, 20, 25, 30,
  • fragments, derivatives, or analogs of the polypeptides of SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81 can be substituted with one or more conserved or non-conserved amino acid residues.
  • an amino acid residue will be substiuted with a conserved amino acid residue.
  • polypeptides, fragments, derivatives, or analogs thereof will have a polypeptide sequence at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the polypeptide sequence shown in SEQ ID NOs:9, 13, 17, 19, 23, 27, 31, 35, 37, 41, 45, 49, 53, 57, 63, 67, 77, or 81 and will comprise functional proteins or enzymes.
  • the native function of the polypeptide will be retained by any such substitutions.
  • amino acid substitutions or mutations are preferably of a minor nature, such as conservative amino acid substitutions that do not significantly affect the folding or activity of the protein.
  • number of amino acid substitutions a skilled artisan would make depends on many factors, including those described herein. Generally, the number of substitutions for any given polypeptide will not be more than about 100, 90, 80, 70, 60, 50, 40, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 5, 6, 4, 3, 2, or 1.
  • isolated polypeptide implies that a polypeptide is removed from its native environment. Thus, a polypeptide produced and/or contained within a recombinant cultured host cell is considered isolated for purposes described herein. Further, “isolated polypeptides” are polypeptides that have been purified, partially or substantially, from a recombinant cultured host.
  • Polypeptides having an amino acid sequence of an indicated percent identity to a reference amino acid sequence can be determined using methods, including computer-assisted methods. Polypeptide amino acid sequences are examined and compared just as are the nucleotide sequences in the foregoing discussion. One of skill in the art will recognize that such concepts as the molecular endpoints discussed for polynucleotides will have direct analogs when considering the corresponding use of such methods and programs for polypeptide analysis.
  • Another embodiment described herein is a method for identifying an auxinic herbicide tolerant plant, or plant part thereof, the method comprising: (a) providing biological material from a plant comprising the DART trait; (b) performing PCR, hybridization testing, or sequencing of said nucleic acid in said biological material to determine if said plant comprises the DART trait; and (c) identifying, based on the results of step (b), that the plant comprises the DART trait.
  • the plant comprises: (i) any one of lines DT-01 Cyc2 BNS4, DT-01
  • Ovaries were surface sterilized in 70% ethanol for 2-3 minutes followed by 10% commercial (Clorox) bleach for 10 minutes. Ovaries were then rinsed 3 times with sterile distilled water, each ovary was cut at the base to remove the pedicel, and cultured upright on BOCBN medium (Table 25).
  • peduncle cultures were initiated from inflorescences of 22, Fl plants in an effort to stimulate chromosomal recombination.
  • the chosen peduncles had closed flower buds and few open flowers. Only the youngest, soft, green tissue was used. Flower buds and pedicels were discarded.
  • the peduncles were surface sterilized with 70% ethanol for 5 minutes, followed by 10% commercial (Clorox) bleach for 10 minutes.
  • the peduncles were rinsed three times with sterile distilled water and the damaged ends were removed. The peduncles were then cut into explants approximately 5 mm long and were cultured horizontally on BPC-BAP medium (Table 25).
  • An in vitro dicamba kill curve was developed for screening seedlings and BC1 plants.
  • Seed of dicamba susceptible (B. napus line BN02 and S. arvensis S line) and resistant (S. arvensis saBC8R and saParR) lines were surface sterilized with 70% ethanol for 2 minutes, then with 30% commercial (Clorox) bleach for 10 minutes, followed by three rinses with sterile distilled water. Seeds were placed on M-LS-513 medium in PlantCons and were allowed to germinate at 23 °C and 16 hours light. When seedlings were 14-21 days old, M-LS-504C medium was melted and mixed with various amounts of dicamba solution to final concentrations ranging from 0-200 mg/L.
  • the dicamba-504C medium combination was then poured on top of the existing M-LS-513 medium, allowed to harden, and the PlantCons were placed back in the growth chamber. After 2-3 weeks of exposure to dicamba, the plants were evaluated. A level of 2.5 mg/L dicamba was determined to give the best differential response (Figure 1).
  • TopazxSa-C LAB 121208 Topaz x DT-01 SA2-R looks normal NT S. arvensis DT lines produced many trichomes on the young leaves and stems, while plants from the Brassica lines had very few trichomes. Thirty-three of the 37 putative Fl plants produced from embryo rescue also displayed the S. arvensis trichome trait ( Figure 2). Leaf tissue was collected from 26 of the Fl plants as well as the parent lines and sent to DNA Landmark for SNP analysis ( Figure 3). The SNP analysis confirmed that the presence of trichomes was correlated with true Fl hybrids.
  • Peduncle culture was also initiated from 22 Fl hybrid plants in an effort to stimulate chromosomal recombination. Christey and Earle, (1991). Plants were regenerated from 12 Fl hybrid lines.
  • BC N F I The protocol used to generate NILs is presented in Figure 4.
  • a total of 8 backcrosses were performed, i.e., BC N F I , where N equals the number of backcrosses.
  • the homozygous recessive S (rr) was used as the recurrent as well as the seed parent, and BC N F 1 progeny were evaluated for response to picloram (100 g ai/ha).
  • Survivors (genotype Rr) were used as the pollen parent for the next BC generation.
  • Individual BCsFi survivors of the picloram treatment were designated BCsFi/A, where A indicates a randomly assigned number for each BCgFi survivor.
  • Plant BCgFi/l was self-pollinated to generate BCgF 2 seed.
  • BCgF 3 seed families (-40-60 seeds/family), designated BCgF 3 /B, where B indicates a randomly assigned number for each BCgF 2 plant.
  • Twenty individual BCgF 3 families were raised and screened with picloram (20 to 30 seedlings per family), to determine the segregation ratio of the R and r alleles and the genotype of the BC 8 F 2 /B plants. Homozygous RR and rr BC 8 F 3 /B families were chosen as NILs.
  • At least four plants each of homozygous R and S NILs were treated with picloram (100 g ai/ha, as above) at the four-leaf stage, and as many of each line were untreated to determine the dry weights of roots and shoots 6 weeks after planting. Dry weights were obtained by rinsing with water and drying roots and shoots at 68 °C for 48 h before weighing. Silique number and seed yield were also determined from four other untreated plants of homozygous R and S NILs allowing them to open pollinate naturally.
  • a morphological trait (serrated leaf margin of first true leaf) associated with auxinic herbicide-resistance in wild mustard was observed. This morphological trait was tested for linkage to the auxinic herbicide-resistance gene. Parental R and S, NIL R and S (30 in each), as well as 75 BC 4 F 1 progeny were assessed for this characteristic 21 DAP (days after planting), and first true leaf areas measured 31 DAP. All seedlings were scored as serrated or smooth before treatment with picloram to determine the genotype at the auxinic herbicide resistance locus of the BC 4 F 1 plants.
  • Table 30 Segregation of resistant (R) and susceptible (S) plants among BC 8 F 3 families (raised from self-pollinated BC 8 F 2 ) following treatment with 100 g ai/ha picloram 1 .
  • NILs near-isogenic lines
  • auxinic herbicide-R and -S gene variants were developed in wild mustard using repeated- backcrosses. These lines were used in growth room studies to examine plant biomass and seed yield parameters that contribute to plant fitness.
  • a morphological marker (leaf shape) was identified that was closely linked to the auxinic herbicide resistance gene in this weed species.
  • R and S wild mustard biotypes showed differences with regard to the shape of first true leaves (Figure 5).
  • the R biotype plants exhibited a serrated leaf margin whereas the S parental plants had a smooth margin.
  • the leaf area of first true leaves of the plants was also significantly greater for S than R ( Figure 6).
  • First true leaves of BC 4 F 1 (75 plants) were measured for leaf area before picloram treatment, then scored for picloram resistance (Table 31).
  • ⁇ 2 is set for the probability of leaf area unlinked to the resistance locus; the results ( ⁇ 2 37.9, degrees of freedom: 3; p ⁇ 0.001) suggests that the gene loci that control leaf morphology and auxinic herbicide resistance are linked.
  • the serrated leaf margin showed perfect correlation (100%) to the smaller leaf area.
  • the segregation of leaf shape and auxinic herbicide resistance trait confirmed that the smaller leaf area/serrated leaf trait is linked and in phase with the R allele, and demonstrated that the gene controlling leaf shape and the gene encoding the DART auxinic herbicide resistance trait are separated by a distance of 14.6 centimorgan (cM) in the wild mustard genome (Table 31).
  • cM centimorgan
  • the field data presented shows auxinic herbicide-tolerance of two plant lines: a Sinapis arvensis line (DT-01 SA2-R) and a Brassica napus line (DT-01 BC4Bn#13-l).
  • the auxinic herbicide-tolerance trait was transferred from the Sinapis arvensis (DT-01 SA2-R) line into the Brassica napus (DT-01 BC4Bn#13-l) line by interspecific crossing.
  • the DT-01 BC4Bn#13-l line was selfed two generations and then subjected to a caged seed increase.
  • the DT-01 SA2-R line and the check lines were also increased in cages in the field. Seed from these caged field productions were used to conduct an herbicide tolerance trial in North Dakota, USA.
  • the three-repetition trial used a randomized split block design.
  • the trial was designed to measure the percent phytotoxicity (% crop injury) observed on the Sinapis arvensis DT-01 SA2- R line versus its non-herbicide-tolerant comparator line, Sinapis arvensis, and the Brassica napus DT-01 BC4Bn#13-l line versus its non-herbicide-tolerant comparator line, Westar.
  • the herbicide used for this study was Clarity (dicamba) with 0.25% NIS (non-ionic surfactant).
  • Three treatment rates of dicamba were applied at the 3-4 leaf stage:
  • the percent phytotoxicity (% crop injury) ratings were transposed into Herbicide Tolerance ratings by subtracting the % crop injury rating (Table 33) for each line from 100% to obtain the percent herbicide tolerance, and then calculating the percent herbicide tolerance present in the target lines, DT-01 BC4Bn#13-l and DT-01 SA2-R, as a function of their non- herbicide tolerant checks (Table 34).
  • auxinic herbicide-treated plants terms such as "without significantly inhibiting the growth" of a plant, refer to plants that exhibit about 50% or less phytotoxicity, as measured 36 days after treatment with an auxinic herbicide, as compared to that exhibited by plants of the corresponding wild-type variety, i.e. the same variety lacking the auxinic herbicide tolerant DART trait found in the herbicide tolerant plant.
  • Table 33 when treated with dicamba at rates of 240 and 480 g ai/ha, the herbicide tolerant B. napus plants exhibit about 56% or less of the phytotoxicity of the corresponding Westar Check line.
  • the mean percent herbicide tolerance was also calculated to show that certain genotypes of Brassica napus have a natural tolerance level to dicamba herbicide.
  • the level of tolerance contributed by the auxinic herbicide -tolerance trait in both DT-01 SA2-R and the Brassica napus DT-01 BC4Bn#13-l lines was significantly higher than the background tolerances observed in their non-herbicide tolerant counterparts (Checks in Table 34).
  • dicamba was applied to plants of the Brassica napus DT-01 BC4Bn#13-l line as well as to the non-herbicide-tolerant comparator line, each at the 2-3 leaf stage, at a rate equivalent to 240 g ai/ha dicamba in order to further determine plant injury.
  • non-resistant Brassica plants of the comparator line showed significant visible injury, defined as (a) leaf epinasty and stem epinasty or (b) leaf epinasty and stem swelling, each in conjunction with a delay in maturity/maturation of more than 2 days.
  • plants of line Brassica napus DT-01 BC4Bn#13-l showed no significant visible injury at 36 days.
  • NILs Near Isogenic-Lines
  • NILs with auxinic herbicide-R and -S were identified upon screening (picloram 100 g ai/ha) BC8F3 families.
  • marker analysis was performed using genomic DNA isolated from wild mustard R and S plants chosen from BC1F1 , BC7F1 , and BC8F1 as well as parental and NILs. Linkage of markers to auxinic herbicide resistance locus was determined by analysis of segregation data from BC progeny.
  • Table 35 shows the tolerance of dicamba-tolerant Sinapis arvensis to auxinic herbicides.
  • Auxinic herbicide-R and -susceptible (S) S. arvensis, B. juncea, and B. rapa were raised from seed.
  • the seeds were sown in 6-inch plastic pots containing Promix (Plant Products, Canada), and placed in a growth chamber having a 16 h photoperiod and 22/15 °C day/night temperature.
  • the light intensity and the relative humidity were maintained at 350 ⁇ m ⁇ 2 s 1 and 65-75%, respectively.
  • Each pot contained one plant and the plants were irrigated when required.
  • Plants were fertilized weekly with 20:20:20 (NPK). When plants were flowering, crosses were performed between auxinic herbicide-R S. arvensis and B. juncea or B.
  • Siliques were dismfected with 70% ethanol for 1 to 2 mill., followed by 20% commercial bleach (sodium hypochlorite, 5.25%) containing three to four drops of Tween* 20 for 15 to 20 min., and subsequently rinsed four to five times with sterile deionized water.
  • the siliques were aseptically cultured in a Petri dish (Fig. 9A) containing 15 mL of either of the following two media: (A). Murashige and Skoog (1962) salts with Gamborg vitamins (1968), sucrose (3%), and 500 mg casein hydrolysate; or (B).
  • the pH of the media was adjusted to 5.8 and 8 g of agar (Difco; micropropagation grade agar was used in all other experiments) was added before autoclaving at 121 °C for 20 min.
  • the siliques were allowed to grow on these media for ⁇ 2 weeks.
  • ovules were excised aseptically from the siliques with a forceps and scalpel and cultured on a Petri dish (2 to 3 ovules per dish; Fig.
  • the sprayer was equipped with a flat- fan nozzle (8002 E) and calibrated to deliver 200 L/ha at 276 kPa.
  • the hybrid plants were classified as R or S by comparing the injury response with those of S. arvensis (dicamba-R) or B. juncea or B. rapa (dicamba-S) seedling response.
  • Susceptibility of plants to dicamba was assessed based on epinasty symptoms (downward curling of leaf and stem tissue) followed by death; R plants were not affected by dicamba application.
  • the fertility of the hybrids was tested by performing reciprocal crosses between hybrids and B. juncea or B. rapa.
  • BCiFi backcross progeny
  • BCiFi seedlings were not screened for dicamba tolerance.
  • BCiFi plants were found male fertile because the pollen from these plants was able to produce BC 2 Fi seed when used as pollen parent. Up to 25 BC 2 Fiseeds were produced , and only 10 were germinated.
  • BC 2 Fi seedlings were at three to four leaf stage of development, they were treated with dicamba (200 g ae/ha).
  • DNA ploidy of hybrids, and backcross progeny was assessed by flow cytometry following the protocol described by Kron et al, (2007). Auxinic herbicide-R S. arvensis, B. juncea and B. rapa were used as controls. Based on DNA content obtained by flow cytometry, DNA ploidy of hybrids as well as backcross progeny was estimated.
  • juncea indicated that these plants were either DNA tetraploids (4 possibly somatic), or aneuploids (plants possessing abnormal number of chromosomes i.e., an extra or missing chromosome; e.g., 4.2x, 4.3*, 4.4x, 4.6* or 4.7*; Table 37).
  • the family Brassicaceae consists of diploids (e.g., B. rapa, B. oleraceae, B. nigra, and S. arvensis etc.) as well as allotetraploids (e.g., B. juncea, B. napus, B. carinata).
  • B. rapa diploids
  • B. oleraceae B. nigra
  • allotetraploids e.g., B. juncea, B. napus, B. carinata
  • arvensis hybrids was achieved. Specifically, the procedure of in vitro culturing of immature siliques 3-5 days after pollination for 2 weeks, followed by excision of embryos/ovules unexpectedly yielded higher number of hybrids in crosses between B. rapa x S. arvensis (32 hybrids) compared to B. juncea x S. arvensis (6 hybrids) (Table 36).
  • dicamba-herbicide-susceptible (dicamba-S) and dicamba-herbicide- tolerant (dicamba-R) Sinapis arvensis plants were sequenced and the genomic sequences were cross-compared for SNPs.
  • dicamba-herbicide-susceptible B. napus and introgressed dicamba-herbicide-tolerant B. napus plant genomes were sequenced and compared for SNPs.
  • CDSs SNPs that occurred in coding sequences (CDSs) and resulted in an expressed amino acid substitution were identified and are shown in Table 38.
  • the CDSs of these genes were annotated for the function of the encoded protein that either: (i) matched a protein listed as a dicamba- binding protein; or (ii) matched a protein listed as an auxin-signal or auxin-transport protein.
  • the resulting group of genes that met either criterion (i) or criterion (ii) comprises the genes and translated polypeptides shown in Table 38.
  • each of the auxin herbicide tolerance trait (-R) genes identified in Table 38 was ascribed as independently providing dicamba herbicide tolerance, i.e. each of these genes encodes a DART auxin herbicide tolerance trait.
  • DART trait genes identified in this analysis 16 were found to correspond to herbicide- susceptible genes. See Table 38, SEQ ID NOs:8, 12, 18, 22, 26, 30, 36, 40, 44, 48, 52, 56, 62, 66, 76, and 80 (Gene Nos. 1-2, 4-7, 9-21). For two of the identified 18 DART trait genes, no corresponding herbicide-susceptible gene has yet been identified.
  • Sar_R_21966.G53064.Tl bodenlos/monopteros SEQ ID NO: 62 SEQ ID NO:63 16 R transcription factor Table 38.
  • S. arvensis Auxin-Herbicide Tolerance Trait Sequences e.g., Dicamba-Tolerance.
  • 1 15S is a second dicamba susceptible gene corresponds to resistant gene 14R.
  • the 18 dicamba resistant, DART trait genes listed in Table 38 were cloned from dicamba-R S. arvensis and used to construct Arabidopsis expression vectors (i.e. the 'R' genes among Gene Nos. 1-21, as shown in Table 38).
  • the herbicide susceptible genes corresponding to the herbicide resistant genes were cloned from dicamba-S S. arvensis and used to construct Arabidopsis expression vectors (i.e., the 'S' genes among Gene Nos. 1-21, as shown in Table 38).
  • the expression vectors were designed for over-expression of these genes. Each of the resulting expression vectors was used to transform A. thaliana plants.
  • BLAST and PSI-BLAST a new generation of protein database search programs.

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Abstract

La présente invention concerne des plantes tolérantes aux herbicides auxiniques, des caractéristiques de tolérance aux herbicides auxiniques et des acides nucléiques et des polypeptides conférant lesdites caractéristiques de résistance aux herbicides auxiniques. L'invention concerne également des procédés permettant de lutter contre la croissance des mauvaises herbes par l'application d'un herbicide auxinique auquel les plantes tolérantes aux herbicides auxiniques décrites ici sont tolérantes.
PCT/IB2014/001106 2013-01-31 2014-01-31 Plantes tolérantes aux herbicides auxiniques WO2014132141A2 (fr)

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CN113899839A (zh) * 2021-10-11 2022-01-07 江南大学 巴戟天中香豆素类化合物的萃取方法
CN114350847A (zh) * 2022-02-11 2022-04-15 四川农业大学 一种鉴定早生茶树的snp位点及其应用

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