Plants Tolerant to HPPD Inhibitor Herbicides
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 Kordia , 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 Kordia hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 387; 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 387, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 195, 219, 220, 221, 222, 234, 237, 238, 239, 240, 249, 299, 329, 372, 373, 367, and 368 with an amino acid set forth in the table below:
Amino acid in
Position in
SEQ ID No. 4
SEQ ID No. 4
Substitutions
Val
195
Thr, Cys, Ala, Gly,
Ile
219
Phe, Tyr, Leu, Val, Ala, Gln, Glu, Asp,
Gly, Thr, Ser, Met, Arg, Lys
Ile
220
Ala, Trp, Leu, Ser, Arg, Lys, His, Asp,
Glu, Pro, Gly, Asn
Phe
222
Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
234
Met, Val
Lys
237
Ala, Val, Leu, Met, Ile, Arg, Gln, Tyr
Met
239
Leu, Val, Ile, Ala
Ser
240
Ala, Thr, Val, Arg, Lys, Glu, Leu, Ile,
Met, His
Ala
372
Glu, Gln, Ser, Val, Phe, Thr
Leu
373
Arg
Ser
221
Glu, Thr, Tyr, Phe, His, Gln, Asn, Gly,
Leu, Met, Val, Arg, Ile
Val
238
Ala, Thr
Pro
249
Ala, Val, Thr, Asn, Ile,
Leu
299
Met, Ile, Asn
Leu
329
Met
Val
367
Any except Pro
Gly
368
Ala, Pro, Val, Thr, Met
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 387.
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 387, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 195, 219, 220, 221, 222, 234, 237, 238, 239, 240, 249, 299, 329, 372, 373, 367, and 368 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 1164, or the nucleic acid sequence of SEQ ID No. 3 from nucleotide position 400 to nucleotide position 1557.
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 387, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 195, 219, 220, 222, 234, 237, 239, 240, 372, and 373 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 387, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 221, 238, 249, 299, 329, 367, and 368 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 387, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 221, 238, 249, 299, 329, 367, and 368 with the amino acid set forth in the following table:
Amino acid in
Position in SEQ ID
SEQ ID No. 4
No. 4
Substitutions
Ser
221
Glu, Thr, Tyr, Arg
Val
238
Ala
Pro
249
Ala, Val, Thr
Leu
299
Met
Leu
329
Met
Val
367
Ile, Ala, Val, Leu, Lys
Gly
368
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. 15, SEQ ID No. 16, SEQ ID No. 17, SEQ ID No. 18, SEQ ID No. 19, SEQ ID No. 20, SEQ ID No. 21, SEQ ID No. 22, and SEQ ID No. 23.
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 Kordia hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 387; 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 387, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 195, 219, 220, 221, 222, 234, 237, 238, 239, 240, 249, 299, 329, 372, 373, 367, and 368 with an amino acid set forth in the table below:
Amino acid in
Position in
SEQ ID No. 4
SEQ ID No. 4
Substitutions
Val
195
Thr, Cys, Ala, Gly, Phe, Tyr, Leu, Val,
Ala, Gln, Glu, Asp, Gly, Thr, Ser, Met,
Arg
Ile
219
Lys
Ile
220
Ala, Trp, Leu, Ser, Arg, Lys, His, Asp,
Glu, Pro, Gly, Asn
Phe
222
Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
234
Met, Val
Lys
237
Ala, Val, Leu, Met, Ile, Arg, Gln, Tyr
Met
239
Leu, Val, Ile, Ala
Ser
240
Ala, Thr, Val, Arg, Lys, Glu, Leu, Ile,
Met, His
Ala
372
Glu, Gln, Ser, Val, Phe, Thr
Leu
373
Arg
Ser
221
Glu, Thr, Tyr, Phe, His, Gln, Asn, Gly,
Leu, Met, Val, Arg, Ile
Val
238
Ala, Thr
Pro
249
Ala, Val, Thr, Asn, Ile,
Leu
299
Met, Ile, Asn
Leu
329
Met
Val
367
Any except Pro
Gly
368
Ala, Pro, Val, Thr, Met
34 . The chimeric gene of claim 33 , wherein the coding sequence comprises the nucleic acid sequence encoding a Kordia HPPD having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 387.
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 387, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 195, 219, 220, 221, 222, 234, 237, 238, 239, 240, 249, 299, 329, 372, 373, 367, and 368 with an amino acid set forth in the table.
36 . A vector comprising the chimeric gene of claim 33 .