EP0946737A2 - Herbicide resistant plants - Google Patents

Herbicide resistant plants

Info

Publication number
EP0946737A2
EP0946737A2 EP97910550A EP97910550A EP0946737A2 EP 0946737 A2 EP0946737 A2 EP 0946737A2 EP 97910550 A EP97910550 A EP 97910550A EP 97910550 A EP97910550 A EP 97910550A EP 0946737 A2 EP0946737 A2 EP 0946737A2
Authority
EP
European Patent Office
Prior art keywords
polynucleotide
herbicide
plants
encoding
gst
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP97910550A
Other languages
German (de)
English (en)
French (fr)
Inventor
Paul Anthony Zeneca Agrochemicals THOMPSON
Mary Elizabeth Zeneca Agrochemicals KNIGHT
Ian Zeneca Agrochemicals JEPSON
Paul Graham Zeneca Agrochemicals THOMAS
Timothy Robert Zeneca Agrochemicals HAWKES
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Syngenta Ltd
Original Assignee
Zeneca Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB9623248.3A external-priority patent/GB9623248D0/en
Priority claimed from GBGB9625957.7A external-priority patent/GB9625957D0/en
Priority claimed from GBGB9703855.8A external-priority patent/GB9703855D0/en
Application filed by Zeneca Ltd filed Critical Zeneca Ltd
Priority to EP06000993A priority Critical patent/EP1728868A2/en
Publication of EP0946737A2 publication Critical patent/EP0946737A2/en
Withdrawn legal-status Critical Current

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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/001—Oxidoreductases (1.) acting on the CH-CH group of donors (1.3)
    • 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
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    • 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
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    • 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
    • C12N15/8275—Glyphosate
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    • 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
    • C12N15/8277—Phosphinotricin
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
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    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
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    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0077—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with a reduced iron-sulfur protein as one donor (1.14.15)
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    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0089—Oxidoreductases (1.) acting on superoxide as acceptor (1.15)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • C12N9/1088—Glutathione transferase (2.5.1.18)
    • C—CHEMISTRY; METALLURGY
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • C12N9/1092—3-Phosphoshikimate 1-carboxyvinyltransferase (2.5.1.19), i.e. 5-enolpyruvylshikimate-3-phosphate synthase
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/93—Ligases (6)

Definitions

  • the invention still further provides plant material which comprises nucleic acid sequences comprising regions encoding at least two proteins capable of conferring upon the material resistance to at least two herbicides, with the provisos that the material that the material does not contain a polynucleotide which encodes a fusion protein comprising only a 5-enol-pyruvyl-3-phosphoshikimate synthetase (EPSPS) and a glutathione S transferase (GST); (ii) that the material does not contain a polynucleotide which comprises only regions encoding superoxide dismutase (SOD) and glutathione S transferase (GST); (iii) that the material does not contain a polynucleotide which comprises only regions encoding GST and phosphinothricin acetyl transferase (PAT); and (iv). that when the plant from which the material is derived is sugar beet, the herbicide resistance or tolerance conferring genes which
  • the invention also includes a method of selectively controlling weeds in a field comprising weeds and crop plants, wherein the crop plants comprise (i) a polynucleotide comprising at least a first region encoding a first protein capable of conferring on a plant, or tissue comprising it, resistance or tolerance to a first herbicide, and a second region encoding a second protein likewise capable of conferring resistance to a second herbicide, with the provisos (i) that the polynucleotide does not encode a fusion protein comprising only a 5- enol-pyruvyl-3-phosphoshikimate synthetase (EPSPS) and a glutathione S transferase (GST); (ii) that the polynucleotide does not comprise only regions encoding superoxide dismutase (SOD) and glutathione S transferase (GST); (iii) that the polynucleotide does not comprise only regions encoding
  • D ⁇ A is isolated and used to transform Agrobacterium tumefaciens LBA4404 to kanamycin resistance again using standard procedures.
  • Potato and tomato tissue is subjected to Agrobacterium-mediated transformation using standard procedures.
  • Transformed shoots are regenerated from kanamycin resistant callus.
  • Shoots are rooted on MS agar containing kanamycin.
  • Surviving rooted explants are re-rooted to provide about 80 kanamycin resistant transformed tobacco plants.
  • the presence of the 4-HPPD gene (using pre-existing EDIT primers) is verified by PCR. A substantial number of PCR positive plants are selected for further analysis.
  • Probes are excised from plasmids, used for Agrobacterium transformation, containing the GST-27 gene by digestion with EcoRI. The probe is labelled
  • This sequence is placed under expression control of an enhanced 35S promoter and ⁇ OS terminator with an Omega fragment being positioned 5' of the protein encoding regions and isolated as an EcoRI Hind III fragment which is cloned into pJRIi to give dicot vector 2.
  • the calli which contain the GST-27 expression cassette are transferred to plant regeneration media and maize plants are recovered.
  • the transformed maize plants are confirmed - by Western blots of total protein extracts from leaves - to constitutively express the GST gene at high levels.
  • Such plants are cross pollinated with an elite maize inbred line and seed is recovered.
  • To confirm enhanced tolerance of the plants to the herbicide acetochlor the said seeds are planted in soil to which has been applied between 2,000 and 8,000 grams per hectare of the herbicide. The seeds are allowed to germinate and grow for 7 days after which time a sample of the resultant seedlings is assessed for damage caused the chemical and compared to the seedlings (if any) which result from non-transgenic seed sown under identical conditions.
  • Plasmid 1 The vector pUBl is digested with Kpnl and a Kpnl -Notl -Kpnl linker inserted, (sequence 5' CAT TTG CGG CCG CAA ATG GTA C 3 - SEQ ID NO. 17).
  • An EcoRl -Notl -EcoRl linker (5' AAT TCA TTT GCG GCC GCA AAT G 3' (SEQ ID No.18) is inserted into the EcoRl site of DVl-pUC.
  • the resulting plasmid is cut with Ncol and the 5' overhang filled using DNA Polymerase 1 Klenow fragment.
  • the linear vector is then digested with Notl and a Notl -blunt fragment isolated.
  • DV3 # 43 B (a glyphosate resistant line comprising the EPSPS and GOX genes - see Example 4) is cross pollinated onto homozygous ET4-208 and vice versa in the usual way. Seed is collected and used in herbicide trials similar to those described above. The tobacco seed is sown in rows in small units the day before treatment. Each compound is formulated in 4% JF5969 (apart from Racer which was a commercial formulation) and sprayed onto the units in the track sprayer at 200 1 ha. The test is assessed at 13, 20 and 27 DAT. Seedlings that are tolerant to bleaching herbicides are transferred after the final assessment into fresh John Innes 111 compost in 3" pots. After two weeks they are subjected to glyphosate herbicide applied at 500 and 800 g/ha. Scoring is performed 14 and 28 DAT. The resultant plants are resistant to both classes of herbicide, and the resistance is inherited in a Mendelian manner.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Plant Pathology (AREA)
  • Botany (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Catching Or Destruction (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
EP97910550A 1996-11-07 1997-10-31 Herbicide resistant plants Withdrawn EP0946737A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP06000993A EP1728868A2 (en) 1996-11-07 1997-10-31 Herbicide resistant plants

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
GBGB9623248.3A GB9623248D0 (en) 1996-11-07 1996-11-07 Herbicide resistant plants
GB9623248 1996-11-07
GB9625957 1996-12-13
GBGB9625957.7A GB9625957D0 (en) 1996-12-13 1996-12-13 Improvements in or relating to organic compounds
GB9703855 1997-02-25
GBGB9703855.8A GB9703855D0 (en) 1997-02-25 1997-02-25 Improvements in or relating to organic compounds
PCT/GB1997/002996 WO1998020144A2 (en) 1996-11-07 1997-10-31 Herbicide resistant plants

Related Child Applications (1)

Application Number Title Priority Date Filing Date
EP06000993A Division EP1728868A2 (en) 1996-11-07 1997-10-31 Herbicide resistant plants

Publications (1)

Publication Number Publication Date
EP0946737A2 true EP0946737A2 (en) 1999-10-06

Family

ID=27268575

Family Applications (2)

Application Number Title Priority Date Filing Date
EP06000993A Withdrawn EP1728868A2 (en) 1996-11-07 1997-10-31 Herbicide resistant plants
EP97910550A Withdrawn EP0946737A2 (en) 1996-11-07 1997-10-31 Herbicide resistant plants

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP06000993A Withdrawn EP1728868A2 (en) 1996-11-07 1997-10-31 Herbicide resistant plants

Country Status (14)

Country Link
EP (2) EP1728868A2 (xx)
JP (1) JP2001503625A (xx)
KR (1) KR20000053140A (xx)
CN (1) CN1236394A (xx)
AU (1) AU4789597A (xx)
BG (1) BG103462A (xx)
BR (1) BR9712695A (xx)
CA (1) CA2269666A1 (xx)
CZ (1) CZ161699A3 (xx)
IL (1) IL129707A0 (xx)
NZ (1) NZ335101A (xx)
PL (1) PL333131A1 (xx)
SK (1) SK60999A3 (xx)
WO (1) WO1998020144A2 (xx)

Families Citing this family (56)

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US6069115A (en) * 1997-11-12 2000-05-30 Rhone-Poulenc Agrochimie Method of controlling weeds in transgenic crops
US6492578B1 (en) 1998-07-10 2002-12-10 Calgene Llc Expression of herbicide tolerance genes in plant plastids
JP2002520024A (ja) * 1998-07-10 2002-07-09 カルジーン エルエルシー 植物色素体における除草剤耐性遺伝子の発現
DE19836659A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Baumwollkulturen
DE19836684A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Reiskulturen
ES2405266T3 (es) 1998-08-13 2013-05-30 Bayer Cropscience Ag Agentes herbicidas para cultivos de maíz tolerantes o resistentes
DE19836700A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Getreidekulturen
DE19836660A1 (de) 1998-08-13 2000-02-17 Hoechst Schering Agrevo Gmbh Herbizide Mittel für tolerante oder resistente Sojakulturen
AU5779499A (en) * 1998-08-28 2000-03-21 Pioneer Hi-Bred International, Inc. Organelle targeting sequences
AU2924800A (en) * 1999-03-03 2000-09-21 Syngenta Limited Use of glutathione-s-transferase to increase stress tolerance in plants
US20020079846A1 (en) * 2000-01-18 2002-06-27 Facchini Peter James Novel glutathione-S-transferase nucleic acids and polypeptides and methods of use thereof
US7083967B1 (en) 2000-06-27 2006-08-01 Basf Corporation Cyanobacterial nucleic acid fragments encoding proteins useful for controlling plant traits via nuclear or plastome transformation
AU2001271486B2 (en) * 2000-06-27 2006-11-09 Basf Plant Science Gmbh Cyanobacterial nucleic acid fragments encoding proteins useful for controlling plant traits via nuclear or plastome transformation
AU2002214158B2 (en) 2000-12-07 2007-01-18 Syngenta Limited Plant derived hydroxy phenyl pyruvate dioxygenases (HPPD) resistant against triketone herbicides and transgenic plants containing these dioxygenases
CA2928061C (en) 2001-10-19 2018-10-16 Sumitomo Chemical Company, Limited Weed controller metabolism proteins, genes thereof and use of the same
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EP1728868A2 (en) 2006-12-06
WO1998020144A2 (en) 1998-05-14
CZ161699A3 (cs) 1999-08-11
JP2001503625A (ja) 2001-03-21
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CN1236394A (zh) 1999-11-24
CA2269666A1 (en) 1998-05-14

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