US2015159145A1PendingUtilityA1

Plants Tolerant to HPPD Inhibitor Herbicides

Assignee: BAYER CROPSCIENCE AGPriority: Dec 23, 2009Filed: Sep 29, 2014Published: Jun 11, 2015
Est. expiryDec 23, 2029(~3.4 yrs left)· nominal 20-yr term from priority
C12N 9/0069C12N 15/8274C12Y 113/11027
58
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Claims

Abstract

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.

Claims

exact text as granted — not AI-modified
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 .

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