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 Euryarchaeota belonging to the family Picrophilaceae, 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 Picrophilus torridus hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 368; 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 368, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 177, 201, 202, 203, 204, 216, 219, 220, 221, 222, 230, 280, 310, 348, 349, 353, and 354 with an amino acid set forth in the table below:
Position
Amino acid in
in SEQ ID
SEQ ID No. 4
No. 4
Substitutions
Val
177
Thr, Cys, Ala, Gly
Leu
201
Phe, Tyr, Ile, Val, Ala, Gln, Glu, Asp, Gly,
Thr, Ser, Met, Arg, Lys
Ile
202
Ala, Trp, Leu, Ser, Arg, Lys, His, Asp,
Glu, Pro, Gly, Asn
Thr
203
Glu, Ser, Tyr, Phe, His, Gln, Asn, Gly,
Leu, Met, Val, Arg, Ile
Phe
204
Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
216
Met, Val
Lys
219
Ala, Val, Leu, Met, Ile, Arg, Gln, Tyr
Val
220
Ala, Thr
Val
221
Leu, Met, Ile, Ala
Lys
222
Ala, Ser, Thr, Val, Arg, Glu, Leu, Ile,
Met, His
Pro
230
Ala, Val, Thr, Asn, Ile,
Leu
280
Met, Ile, Asn
Leu
310
Met
Asn
348
Any except Pro
Gly
349
Ala, Pro, Val, Thr, Met
Ala
353
Glu, Gln, Ser, Val, Phe, Thr
Leu
354
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 368.
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 368, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 177, 201, 202, 203, 204, 216, 219, 220, 221, 222, 230, 280, 310, 348, 349, 353, and 354 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 1107, or the nucleic acid sequence of SEQ ID No. 3 from nucleotide position 396 to nucleotide position 1503.
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 368, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 177, 201, 202, 204, 216, 219, 221, 222, 353, and 354 with the amino acid 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 368, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 203, 220, 230, 280, 310, 348, and 349 with the amino acid 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 368, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 203, 220, 230, 280, 310, 348, and 349 with the amino acid set forth in the following table:
Position
Amino acid in
in SEQ ID
SEQ ID No. 4
No. 4
Substitutions
Thr
203
Glu, Ser, Arg, Tyr
Val
220
Ala
Pro
230
Ala, Val, Thr
Leu
280
Met
Leu
310
Met
Asn
348
Ile, Ala, Val, Leu, Lys
Gly
349
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 Picrophilus torridus hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 368; 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 368, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 177, 201, 202, 203, 204, 216, 219, 220, 221, 222, 230, 280, 310, 348, 349, 353, and 354 with an amino acid set forth in the table below:
Position
Amino acid in
in SEQ ID
SEQ ID No. 4
No. 4
Substitutions
Val
177
Thr, Cys, Ala, Gly
Leu
201
Phe, Tyr, Ile, Val, Ala, Gln, Glu, Asp, Gly,
Thr, Ser, Met, Arg, Lys
Ile
202
Ala, Trp, Leu, Ser, Arg, Lys, His, Asp,
Glu, Pro, Gly, Asn
Thr
203
Glu, Ser, Tyr, Phe, His, Gln, Asn, Gly,
Leu, Met, Val, Arg, Ile
Phe
204
Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
216
Met, Val
Lys
219
Ala, Val, Leu, Met, Ile, Arg, Gln, Tyr
Val
220
Ala, Thr
Val
221
Leu, Met, Ile, Ala
Lys
222
Ala, Ser, Thr, Val, Arg, Glu, Leu, Ile,
Met, His
Pro
230
Ala, Val, Thr, Asn, Ile,
Leu
280
Met, Ile, Asn
Leu
310
Met
Asn
348
Any except Pro
Gly
349
Ala, Pro, Val, Thr, Met
Ala
353
Glu, Gln, Ser, Val, Phe, Thr
Leu
354
Arg
34 . The chimeric gene of claim 33 , wherein the coding sequence comprises the nucleic acid sequence encoding a Picrophilus torridus HPPD having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 368.
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 368, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 177, 201, 202, 203, 204, 216, 219, 220, 221, 222, 230, 280, 310, 348, 349, 353, and 354 with an amino acid set forth in the table.
36 . A vector comprising the chimeric gene of claim 33 .