Engineering single-gene-controlled staygreen potential into plants
AssigneesPioneer Hi-Bred International, Inc., The Regents of the University of California
InventorsDaniel Gallie, Robert Meeley, Todd Young
Life of this patent
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Abstract
The enzymes of the ACC synthase family are used in producing ethylene. Nucleotide and polypeptide sequences of ACC synthases are provided along with knockout plant cells having inhibition in expression and/or activity in an ACC synthase and knockout plants displaying a staygreen phenotype, a male sterility phenotype, or an inhibition in ethylene production. Methods for modulating staygreen potential in plants, methods for modulating sterility in plants, and methods for inhibiting ethylene production in plants are also provided.
Claims60
An isolated or recombinant knockout plant cell comprising at least one disruption in at least one endogenous ACC synthase gene, wherein the disruption inhibits expression or activity of at least one ACC synthase protein compared to a corresponding control plant cell lacking the disruption, wherein the at least one endogenous ACC synthase gene comprises a nucleic acid sequence, or complement thereof, comprising at least about 70% sequence identity to SEQ ID NO:1 (gACS2), SEQ ID NO:2 (gACS6), or SEQ ID NO:3 (gACS7).
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4-12. (canceled)
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The plant of claim 22, wherein the plant is Zea mays, wheat, rice, sorghum, barley, oat, lawn grass, rye, soybean, Brassica, or sunflower.
20-21. (canceled)
A knockout plant comprising a staygreen potential phenotype, the staygreen potential phenotype resulting from a disruption in at least one endogenous ACC synthase gene, wherein the disruption comprises one or more transposons or one or more point mutations, and wherein the disruption inhibits expression or activity of at least one ACC synthase protein compared to a corresponding control plant.
24-28. (canceled)
A transgenic knockout plant comprising a staygreen potential phenotype, said staygreen potential phenotype resulting from at least one introduced transgene which inhibits ethylene synthesis, wherein said at least one introduced transgene comprises a nucleic acid sequence encoding at least one ACC synthase, which nucleic acid sequence comprises at least about 85% sequence identity to SEQ ID NO:1 (gACS2), SEQ ID NO:2 (gACS6), SEQ ID NO:3 (gACS7), SEQ ID NO:4 (cACS2), SEQ ID NO:5 (cACS6), SEQ ID NO:6 (cACS7) or SEQ ID NO:10 (CCRA178R), or a subsequence thereof, or a complement thereof, and is in a configuration that modifies a level of expression or activity of the at least one ACC synthase.
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A transgenic plant comprising a staygreen potential phenotype, said staygreen potential phenotype resulting from at least one introduced transgene which inhibits ethylene synthesis, wherein said at least one introduced transgene comprises a nucleic acid sequence encoding a subsequence of at least one ACC synthase, which at least one ACC synthase comprises at least about 85% sequence identity to SEQ ID NO:7 (pACS2), SEQ ID NO:8 (pACS6), SEQ ID NO.:9 (pACS7), or SEQ ID NO:11 (pCCRA178R), is in an RNA silencing or interference configuration, and modifies a level of expression or activity of the at least one ACC synthase.
An isolated or recombinant polynucleotide comprising a member selected from the group consisting of:
- (a) a polynucleotide, or a complement thereof, comprising at least about 85% sequence identity to SEQ ID NO:1 (gACS2), SEQ ID NO:3 (gACS7), SEQ ID NO:4 (cACS2), SEQ ID NO:6 (cACS7), or SEQ ID NO:10 (CCRA178R), or a subsequence comprising at least about 100 contiguous nucleotides thereof;
- (b) a polynucleotide, or a complement thereof, encoding a polypeptide sequence of SEQ ID NO:7 (pACS2), SEQ ID NO.:9 (pACS7), or SEQ ID NO:11 (pCCRA178R), or a subsequence comprising at least about 50 contiguous amino acids thereof, or a polypeptide sequence at least about 95% identical to SEQ ID NO:7, SEQ ID NO.:9, or SEQ ID NO:11;
- and,
- (c) a polynucleotide, or a complement thereof, that hybridizes under stringent conditions over substantially the entire length of a polynucleotide subsequence comprising at least 100 contiguous nucleotides of SEQ ID NO:1 (gACS2), SEQ ID NO:3 (gACS7), SEQ ID NO:4 (cACS2), SEQ ID NO:6 (cACS7), or SEQ ID NO:10 (CCRA 178R).
A knockout plant comprising a male sterility phenotype, the male sterility phenotype resulting from at least one disruption in at least one endogenous ACC synthase gene, the disruption inhibiting expression or activity of at least one ACC synthase protein compared to a corresponding control plant, wherein the at least one endogenous ACC synthase gene comprises a nucleic acid sequence, or complement thereof, comprising at least about 70% sequence identity to SEQ ID NO:1 (gACS2), SEQ ID NO:2 (gACS6), or SEQ ID NO:3 (gACS7).
52-56. (canceled)
A transgenic knockout plant comprising a male sterility phenotype, said male sterility phenotype resulting from at least one introduced transgene which inhibits ethylene synthesis, wherein said at least one introduced transgene comprises a nucleic acid sequence encoding at least one ACC synthase, which nucleic acid sequence comprises at least about 85% sequence identity to SEQ ID NO:1 (gACS2), SEQ ID NO:2 (gACS6), SEQ ID NO:3 (gACS7), SEQ ID NO:4 (cACS2), SEQ ID NO:5 (cACS6), SEQ ID NO:6 (cACS7) or SEQ ID NO:10 (CCRA178R), or a subsequence thereof, or a complement thereof, and is in a configuration that modifies a level of expression or activity of the at least one ACC synthase.
58-59. (canceled)
A method of inhibiting ethylene production in a plant, the method comprising:
- inactivating one or more ACC synthase genes in the plant, wherein the one or more ACC synthase genes encode one or more ACC synthases, wherein at least one of the one or more ACC synthases comprises at least about 85% identity to SEQ ID NO:7 (pACS2), SEQ ID NO:8 (pACS6), SEQ ID NO:9 (pAC7) or SEQ ID NO:11 (pCCRA178R).
61-67. (canceled)
The method of claim 67, wherein the at least one polynucleotide sequence is introduced by electroporation, micro-projectile bombardment or Agrobacterium-mediated transfer.
The method of claim 67, wherein the polynucleotide is linked to the promoter in a sense orientation.
The method of claim 67, wherein the polynucleotide is linked to the promoter in an antisense orientation.
The method of claim 67, wherein the polynucleotide is configured for RNA silencing or interference.
The method of claim 67, wherein the promoter is a tissue-specific promoter or an inducible promoter.
A method for modulating staygreen potential in a plant, the method comprising:
- a) selecting at least one ACC synthase gene to mutate, which ACC synthase gene comprises a nucleic acid sequence, or complement thereof, comprising at least about 70% sequence identity to SEQ ID NO:1 (gACS2), SEQ ID NO:2 (gACS6), or SEQ ID NO:3 (gACS7), thereby providing at least one desired ACC synthase gene;
- b) introducing a mutant form of the at least one desired ACC synthase gene into the plant;
- and,
- c) expressing the mutant form, thereby modulating staygreen potential in the plant.
77-87. (canceled)
A kit for modulating staygreen potential in a plant, the kit comprising:
- at least one polynucleotide sequence comprising a nucleic acid sequence, wherein the nucleic acid sequence is at least about 85% identical to SEQ ID NO:1 (gACS2), SEQ ID NO:3 (gACS7), SEQ ID NO:4 (cACS2), SEQ ID NO:6 (cAC7) or SEQ ID NO:10 (CCRA178R), or a complement thereof.
A kit for modulating male sterility in a plant, the kit comprising:
- at least one polynucleotide sequence comprising a nucleic acid sequence, wherein the nucleic acid sequence is at least about 85% identical to SEQ ID NO:1 (gACS2), SEQ ID NO:2 (gACS6), SEQ ID NO:3 (gACS7), SEQ ID NO:4 (cACS2), SEQ ID NO:5 (cACS6), SEQ ID NO:6 (cAC7) or SEQ ID NO:10 (CCRA178R), or a subsequence thereof, or a complement thereof.
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Dependencies are inferred from the claim text.