Plants Tolerant to HPPD Inhibitor Herbicides
The present invention relates to nucleic acid sequences encoding a hydroxyphenylpyruvate dioxygenase (EC 1.13.11.27, abbreviated herein as HPPD) obtained from bacteria belonging to the genus Rhodococcus as well as the proteins encoded thereby, and to a chimeric gene which comprises such nucleic acid sequence, and to the use of such nucleic acid sequences, proteins or chimeric genes for obtaining plants which are tolerant to HPPD inhibitor herbicides.
1 - 16 . (canceled)
17 . A plant, a plant part, a plant cell, or a seed comprising a chimeric gene comprising a coding sequence operably-linked to a plant-expressible promoter, wherein the coding sequence comprises
a) a nucleic acid sequence encoding a Rhodococcus sp. hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401; or
b) a nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 207, 231, 232, 233, 234, 246, 249, 250, 251, 252, 261, 311, 343, 381, 382, 386, and 387 with an amino acid set forth in the table below:
Amino acid
Position
in SEQ
in SEQ
ID No. 4
ID No. 4
Substitutions
Val
207
Thr, Cys, Ala, Gly
Met
231
Phe, Tyr, Ile, Leu, Val, Ala, Gln, Glu,
Asp, Gly, Thr, Ser, Arg, Lys
Ala
232
Ile, Trp, Leu, Ser, Arg, Lys, His, Asp, Glu,
Pro, Gly, Asn
Glu
233
Ser, Thr, Tyr, Phe, His, Gln, Asn, Gly,
Leu, Met, Val, Arg, Ile
Phe
234
Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
246
Met, Val
Lys
249
Ala, Val, Leu, Met, Ile, Arg, Gln, Tyr
Val
250
Ala, Thr
Val
251
Leu, Met, Ile, Ala
Ser
252
Ala, Thr, Val, Arg, Lys, Glu, Leu, Ile,
Met, His
Pro
261
Ala, Val, Thr, Asn, Ile,
Leu
311
Met, Ile, Asn
Leu
343
Met
Ile
381
Any except Pro
Gly
382
Ala, Pro, Val, Thr, Met
Ala
386
Glu, Gln, Ser, Val, Phe, Thr
Leu
387
Arg.
18 . The plant, the plant part, the plant cell, or the seed of claim 17 , wherein the coding sequence comprises the nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401.
19 . The plant, the plant part, the plant cell, or the seed of claim 17 , wherein the coding sequence comprises the nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 207, 231, 232, 233, 234, 246, 249, 250, 251, 252, 261, 311, 343, 381, 382, 386, and 387 with an amino acid as set forth in the table in claim 17 .
20 . The plant, the plant part, the plant cell, or the seed of claim 18 , wherein the coding sequence comprises the nucleotide sequence of SEQ ID No. 1 from nucleotide position 4 to nucleotide position 1203, or the nucleic acid sequence of SEQ ID No. 3 from nucleotide position 400 to nucleotide position 1602.
21 . The plant, the plant part, the plant cell, or the seed of claim 19 , wherein the coding sequence comprises a nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 207, 231, 232, 234, 246, 249, 251, 252, 386, and 387 with the amino acid as set forth in the table.
22 . The plant, the plant part, the plant cell, or the seed of claim 19 , wherein the coding sequence comprises a nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 233, 250, 261, 311, 343, 381, and 382 with the amino acid as set forth in the table.
23 . The plant, the plant part, the plant cell, or the seed of claim 19 , wherein the coding sequence comprises a nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 233, 250, 261, 311, 343, 381, and 382 with the amino acid set forth in the following table:
Amino acid
Position
in SEQ
in SEQ
ID No. 4
ID No. 4
Substitutions
Glu
233
Thr, Arg, Tyr, Ser
Val
250
Ala
Pro
261
Ala, Val, Thr
Leu
311
Met
Leu
343
Met
Ile
381
Ala, Val, Leu, Lys
Gly
382
Ala
24 . The plant, the plant part, the plant cell, or the seed of claim 18 , wherein the chimeric gene further comprises a nucleic acid sequence encoding a transit peptide active in a plant upstream of the HPPD coding sequence, so that a transit peptide/HPPD fusion protein is encoded by the chimeric gene.
25 . The plant, the plant part, the plant cell, or the seed of claim 18 further comprising a chimeric gene encoding a prephenate dehydrogenase (PDH) enzyme.
26 . The plant, the plant part, the plant cell, or the seed of claim 18 further comprising one or more chimeric genes conferring tolerance to
(a) a growth regulator herbicide;
(b) a herbicide inhibiting enzyme, wherein the enzyme is selected from the group consisting of (i) acetolactate synthase, (ii) EPSP synthase, and (iii) glutamine synthase; or
(c) a combination thereof.
27 . The plant, the plant part, the plant cell, or the seed of claim 26 , wherein the growth regulator herbicide is 2,4-D or dicamba.
28 . The plant, the plant part, the plant cell, or the seed of claim 19 , wherein the chimeric gene further comprises a nucleic acid sequence encoding a transit peptide active in a plant upstream of the coding sequence, so that a transit peptide fusion protein is encoded by the chimeric gene.
29 . The plant, the plant part, the plant cell, or the seed of claim 19 further comprising a chimeric gene encoding a prephenate dehydrogenase (PDH) enzyme.
30 . The plant, the plant part, the plant cell, or the seed of claim 19 further comprising one or more chimeric genes conferring tolerance to
(a) a growth regulator herbicide;
(b) a herbicide inhibiting enzyme, wherein the enzyme is selected from the group consisting of (i) acetolactate synthase, (ii) EPSP synthase, and (iii) glutamine synthase; or
(c) a combination thereof.
31 . The plant, the plant part, the plant cell, or the seed of claim 30 , wherein the growth regulator herbicide is 2,4-D or dicamba.
32 . A plant, a plant part, a plant cell, or a seed comprising a chimeric gene comprising a coding sequence operably linked to a plant-expressible promoter, wherein the coding sequence comprises a nucleic acid sequence selected from the group consisting of SEQ ID No. 22, SEQ ID No. 23, SEQ ID No. 24, SEQ ID No. 25, SEQ ID No. 26, SEQ ID No. 27, SEQ ID No. 28, SEQ ID No. 29, SEQ ID No. 30, SEQ ID No. 31, SEQ ID No. 32, SEQ ID No. 33, SEQ ID No. 34, SEQ ID No. 35, SEQ ID No. 36, SEQ ID no. 37, SEQ ID No. 38, and SEQ ID No. 39.
33 . A chimeric gene comprising a coding sequence operably-linked to a plant-expressible promoter, wherein the coding sequence comprises
a) a nucleic acid sequence encoding a Rhodococcus sp. hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401; or
b) a nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 207, 231, 232, 233, 234, 246, 249, 250, 251, 252, 261, 311, 343, 381, 382, 386, and 387 with an amino acid set forth in the table below:
Amino acid
Position
in SEQ
in SEQ
ID No. 4
ID No. 4
Substitutions
Val
207
Thr, Cys, Ala, Gly
Met
231
Phe, Tyr, Ile, Leu, Val, Ala, Gln, Glu,
Asp, Gly, Thr, Ser, Arg, Lys
Ala
232
Ile, Trp, Leu, Ser, Arg, Lys, His, Asp, Glu,
Pro, Gly, Asn
Glu
233
Ser, Thr, Tyr, Phe, His, Gln, Asn, Gly,
Leu, Met, Val, Arg, Ile
Phe
234
Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
246
Met, Val
Lys
249
Ala, Val, Leu, Met, Ile, Arg, Gln, Tyr
Val
250
Ala, Thr
Val
251
Leu, Met, Ile, Ala
Ser
252
Ala, Thr, Val, Arg, Lys, Glu, Leu, Ile,
Met, His
Pro
261
Ala, Val, Thr, Asn, Ile,
Leu
311
Met, Ile, Asn
Leu
343
Met
Ile
381
Any except Pro
Gly
382
Ala, Pro, Val, Thr, Met
Ala
386
Glu, Gln, Ser, Val, Phe, Thr
Leu
387
Arg
34 . The chimeric gene of claim 33 , wherein the coding sequence comprises the nucleic acid sequence encoding a Rhodococcus sp. HPPD having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401.
35 . The chimeric gene of claim 33 , wherein the coding sequence comprises the nucleic acid sequence encoding the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 401, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 207, 231, 232, 233, 234, 246, 249, 250, 251, 252, 261, 311, 343, 381, 382, 386, and 387 with an amino acid as set forth in the table of claim 33 .
36 . A vector comprising the chimeric gene of claim 33 .