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 subfamily Synechococcoideae, 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 Synechococcus sp. hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 350; 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 350, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 145, 169, 170, 171, 172, 181, 184, 185, 186, 187, 196, 244, 281, 327, 328, 332, or 333 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
145
Thr, Cys, Ala, Gly
Leu
169
Phe, Tyr, Ile, Val, Ala, Gln, Glu, Asp, Gly,
Thr, Ser, Met, Arg, Lys
Tyr
170
Ala, Trp, Ile, Leu, Ser, Arg, Lys, His, Asp,
Glu, Pro, Gly, Asn
Arg
171
Glu, Ser, Thr, Tyr, Phe, His, Gln, Asn,
Gly, Leu, Met, Val, Ile
Tyr
172
Phe, Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
181
Met, Val
Val
184
Ala, Leu, Met, Ile, Lys, Arg, Gln, Tyr
Val
185
Ala, Thr
Val
186
Leu, Met, Ile, Ala
Gly
187
Ala, Ser, Thr, Val, Arg, Lys, Glu, Leu, Ile,
Met, His
Ala
196
Pro, Val, Thr, Asn, Ile,
Leu
244
Met, Ile, Asn
Leu
281
Met
Glu
327
Any except Pro
Ala
328
Gly, Pro, Val, Thr, Met
Ala
332
Glu, Gln, Ser, Val, Phe, Thr
Leu
333
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 350.
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 350, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 145, 169, 170, 171, 172, 181, 184, 185, 186, 187, 196, 244, 281, 327, 328, 332, or 333 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 1053, or the nucleic acid sequence of SEQ ID No. 3 from nucleotide position 400 to nucleotide position 1446.
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 350, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 145, 169, 170, 172, 181, 184, 186, 187, 332, and 333 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 382, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 171, 185, 196, 244, 281, 327, and 328 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 350, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 171, 185, 196, 244, 281, 327, and 328 with the amino acid set forth in the following table:
Amino acid
Position
in SEQ
in SEQ
ID No. 4
ID No. 4
Substitutions
Arg
171
Glu, Thr, Ser, Tyr
Val
185
Ala
Ala
196
Pro, Val, Thr
Leu
244
Met
Leu
281
Met
Glu
327
Ile, Ala, Val, Leu, Lys
Ala
328
Gly
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. 18, SEQ ID No. 19, SEQ ID No. 20, SEQ ID No. 21, SEQ ID No. 22, SEQ ID No. 23, SEQ ID No. 24, and SEQ ID No. 25.
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 Synechococcus sp. hydroxyphenylpyruvate dioxygenase (HPPD) having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 350; 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 350, wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 145, 169, 170, 171, 172, 181, 184, 185, 186, 187, 196, 244, 281, 327, 328, 332, or 333 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
145
Thr, Cys, Ala, Gly
Leu
169
Phe, Tyr, Ile, Val, Ala, Gln, Glu, Asp, Gly,
Thr, Ser, Met, Arg, Lys
Tyr
170
Ala, Trp, Ile, Leu, Ser, Arg, Lys, His, Asp,
Glu, Pro, Gly, Asn
Arg
171
Glu, Ser, Thr, Tyr, Phe, His, Gln, Asn,
Gly, Leu, Met, Val, Ile
Tyr
172
Phe, Val, Ile, Ala, Leu, Trp, Met, Gln, His
Leu
181
Met, Val
Val
184
Ala, Leu, Met, Ile, Lys, Arg, Gln, Tyr
Val
185
Ala, Thr
Val
186
Leu, Met, Ile, Ala
Gly
187
Ala, Ser, Thr, Val, Arg, Lys, Glu, Leu, Ile,
Met, His
Ala
196
Pro, Val, Thr, Asn, Ile,
Leu
244
Met, Ile, Asn
Leu
281
Met
Glu
327
Any except Pro
Ala
328
Gly, Pro, Val, Thr, Met
Ala
332
Glu, Gln, Ser, Val, Phe, Thr
Leu
333
Arg
34 . The chimeric gene of claim 33 , wherein the coding sequence comprises the nucleic acid sequence encoding a Blepharisma japonicum HPPD having the amino acid sequence of SEQ ID No. 4 from amino acid position 2 to amino acid position 350.
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 350, and wherein one or more amino acids of SEQ ID No. 4 are substituted at amino acid positions 145, 169, 170, 171, 172, 181, 184, 185, 186, 187, 196, 244, 281, 327, 328, 332, or 333 with an amino acid as set forth in the table of claim 33 .
36 . A vector comprising the chimeric gene of claim 33 .