US2003192072A1PendingUtilityA1
Methods for making plants tolerant to glyphosate and compositions thereof
Est. expiryMar 9, 2020(expired)· nominal 20-yr term from priority
C12N 15/8275C12N 9/1092
59
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Claims
Abstract
The methods and materials disclosed herein are directed to glyphosate herbicide tolerance in plants. In particular, the isolation of a glyphosate resistant EPSP synthase coding sequence and its regulatory elements from Eleusine indica. The coding sequence and regulatory sequences are useful to genetically engineer plants for tolerance to glyphosate herbicide.
Claims
exact text as granted — not AI-modifiedWe claim
1 . A DNA molecule that encodes a naturally occurring glyphosate resistant EPSPS enzyme derived from a glyphosate tolerant plant, wherein the glyphosate resistant EPSPS enzyme has a K m for phosphoenolpyruvate (PEP) of less than 10 μM.
2 . A DNA molecule of claim 1 that encodes a naturally occurring glyphosate resistant EPSPS enzyme derived from a glyphosate tolerant plant, wherein the glyphosate resistant EPSPS enzyme has a K m for PEP of less than 10 μM and the K m for PEP is not more than about twice of the K m for PEP of a naturally occurring glyphosate sensitive EPSPS enzyme derived from a glyphosate sensitive plant.
3 . A DNA molecule of claim 2 , wherein said plant is Eleusine species.
4 . A DNA molecule of claim 1 , wherein said naturally occurring glyphosate resistant EPSPS enzyme is modified by a substitution or a deletion of at least one amino acid in a catalytic domain.
5 . A DNA molecule of claim 4 , wherein said substitution is selected from the group consisting of glycine to alanine 102 and threonine to isoleucine 103 of SEQ ID NO: 7.
6 . A DNA molecule that encodes a naturally occurring glyphosate resistant EPSPS enzyme of SEQ ID NO: 7.
7 . A DNA molecule of claim 6 that encodes a naturally occurring glyphosate resistant EPSPS enzyme of SEQ ID NO: 7, wherein the DNA molecule is substantially homologous to SEQ ID NO: 6.
8 . A recombinant DNA molecule comprising: a promoter that functions in plant cells to cause the production of an RNA sequence, operably linked to; a structural DNA sequence that causes the production of an RNA sequence that encodes an EPSPS enzyme comprising the sequence of SEQ ID NO: 7, operably linked to; a 3′ non-translated region that functions in plant cells to cause the addition of polyadenyl nucleotides to the 3′end of the RNA sequence, wherein the promoter is heterologous with respect to the structural DNA sequence and selected so as to cause sufficient expression of the polypeptide to enhance the glyphosate tolerance of a transgenic plant cell containing said recombinant DNA molecule.
9 . A recombinant DNA molecule of claim 8 , wherein said structural DNA sequence encodes a fusion polypeptide comprising an amino-terminal chloroplast transit peptide and an EPSPS enzyme comprising the sequence of SEQ ID NO: 7.
10 . A method of producing glyphosate tolerant plants comprising the steps of:
a) inserting into the genome of a plant cell a recombinant DNA molecule comprising: a promoter that functions in plant cells to cause the production of a RNA sequence, operably linked to; a structural DNA sequence that caused the production of a RNA sequence that encodes an EPSPS enzyme having the sequence of SEQ ID NO: 7, operably linked to; a 3′ non-translated region that functions in plant cells to cause the addition of polyadenyl nucleotides the 3′end of the RNA sequence; where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the polypeptide to enhance the glyphosate tolerance of a plant cell transformed with the DNA molecule; b) obtaining a transformed plant cell; and c) regenerating from the transformed plant cell a genetically transformed plant which has increased tolerance to glyphosate herbicide.
11 . A method of claim 10 , wherein the structural DNA sequence encodes a fusion polypeptide comprising an amino-terminal chloroplast transit peptide and an EPSPS enzyme comprising the sequence of SEQ ID NO: 7.
12 . A glyphosate tolerant plant cell comprising a recombinant DNA molecule of claim 8 or 9 .
13 . A glyphosate tolerant plant cell of claim 12 selected from the group consisting of corn, wheat, rice, millet, sugarcane, barley, oat, rye, turf grasses, asparagus, soybean, cotton, sugar beet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, forest trees, fruit trees, ornamental annuals, and ornamental perennials.
14 . A glyphosate tolerant plant comprising the plant cells of claim 12 .
15 . A glyphosate tolerant plant of claim 14 selected from the group consisting of corn, wheat, rice, millet, sugarcane, barley, oat, rye, turf grasses, asparagus, soybean, cotton, sugar beet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, forest trees, fruit trees, ornamental annuals, and ornamental perennials.
16 . A recombinant DNA molecule comprising: a promoter that functions in plant cells to cause the production of an RNA sequence, operably linked to; a structural DNA sequence that causes the production of an RNA sequence which encodes an EPSPS enzyme having the sequence of SEQ ID NO: 7, operably linked to; a 3′ non-translated region that functions in plant cells to cause the addition of polyadenyl nucleotides the 3′ end of the RNA sequence, wherein the promoter is homologous with respect to the structural DNA sequence.
17 . A DNA molecule of claim 16 wherein the structural DNA sequence encodes a fusion polypeptide comprising an amino-terminal chloroplast transit peptide and the EPSPS enzyme comprising the sequence of SEQ ID NO: 7.
18 . A glyphosate tolerant transgenic plant cell comprising a DNA molecule of claim 16 or 17 .
19 . A glyphosate tolerant transgenic plant cell of claim 18 selected from the group consisting of corn, wheat, rice, millet, sugarcane, barley, oat, rye, turf grasses, asparagus, soybean, cotton, sugar beet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, forest trees, fruit trees, ornamental annuals, ornamental perennials.
20 . A glyphosate tolerant transgenic plant comprising plant cells of claim 18 .
21 . Glyphosate tolerant transgenic plant of claim 20 selected from the group consisting of corn, wheat, rice, millet, sugarcane, barley, oat, rye, turf grasses, asparagus, soybean, cotton, sugar beet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, forest trees, fruit trees, ornamental annuals, ornamental perennials.
22 . The seed of a glyphosate tolerant transgenic plant of claim 15 .
23 . The seed of a glyphosate tolerant transgenic plant of claim 21 .
24 . A DNA molecule comprising the promoter region located 5′ to the DNA molecule of claim 3 .
25 . A DNA molecule comprising the chloroplast transit peptide coding region located 5′ to the DNA molecule of claim 3 .
26 . A DNA molecule comprising the 3′ untranslated region located 3′ to the DNA molecule of claim 3 .
1
GCGGGCGCGG AGGAGGTGGT GCTGCAGCCC ATCAAGGAGA TCTCCGGCGT
51
CGTGAAGCTG CCGGGGTCCA AGTCGCTCTC CAACCGGATC CTCCTGCTCT
101
CCGCCCTCGC CGAGGGAACA ACTGTGGTGG ATAACCTTTT AAACAGTGAG
151
GACGTCCACT ACATGCTCGG GGCCCTGAAA ACCCTCGGAC TCTCTGTGGA
201
AGCGGACAAA GCTGCCAAAA GAGCGGTAGT TGTTGGCTGT GGTGGCAAGT
251
TCCCAGTTGA GAAGGATGCG AAAGAGGAGG TGCAGCTCTT CTTGGGGAAT
301
GCTGGAACTG CAATGCGATC ATTGACAGCA GCCGTAACTG CTGCTGGAGG
351
AAATGCAACT TATGTGCTTG ATGGAGTGCC AAGAATGCGG GAGAGACCCA
401
TTGGCGACTT GGTTGTCGGA TTGAAACAGC TTGGTGCGGA TGTTGATTGT
451
TTCCTTGGCA CTGACTGCCC ACCTGTTCGT GTCAAGGGAA TCGGAGGGCT
501
ACCTGGTGGC AAGGTTAAGT TATCTGGTTC CATCAGCAGT CAGTACTTGA
551
GTGCCTTGCT GATGGCTGCT CCTTTAGCTC TTGGGGATGT GGAGATTGAA
601
ATCATTGATA AACTGATCTC CATCCCTTAT GTTGAAATGA CATTGAGATT
651
GATGGAGCGT TTTGGCGTGA AAGCAGAGCA TTCTGATAGC TGGGACAGAT
701
TCTACATCAA GGGAGGTCAA AAATACAAGT CCCCTAAAAA TGCCTACGTG
751
GAAGGTGATG CCTCAAGTGC GAGCTATTTC TTGGCTGGTG CTGCAATCAC
801
TGGAGGGACT GTGACTGTTG AAGGTTGTGG CACCACCAGT CTGCAGGGTG
851
ATGTGAAATT TGCCGAGGTA CTCGAGATGA TGGGAGCGAA GGTTACATGG
901
ACTGAAACTA GCGTAACTGT TACCGGTCCA CAACGTGAGC CATTTGGGAG
951
GAAACACCTA AAAGCTATTG ATGTTAACAT GAACAAAATG CCCGATGTCG
1001
CCATGACTCT TGCCGTGGTT GCCCTATTTG CTGATGGCCC AACTGCTATC
1051
AGAGATGTGG CTTCCTGGAG AGTAAAGGAG ACCGAGAGGA TGGTTGCAAT
1101
CCGGACTGAG CTAACAAAGC TGGGAGCGTC GGTCGAGGAA GGACTGGACT
1151
ACTGCATTAT CACACCGCCC GAGAAGCTGA ACGTAACGGC CATCGACACC
1201
TACGATGACC ACAGGATGGC CATGGCCTTC TCCCTCGCCG CCTGCGCCGA
1251
CGTGCCTGTG ACCATCCGGG ACCCCGGCTG CACCCGCAAG ACCTTCCCAG
1301
ACTACTTCGA CGTGCTGAGC ACTTTCGTCA AGAACTAAJoin the waitlist — get patent alerts
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