Alteration of embryo/endosperm size during seed development

Abstract

Isolated nucleic acid fragments and recombinant constructs comprising such fragments for altering embryo/endosperm size during seed development are disclosed along with a method of controlling embryo/endosperm size during seed development in plants.

1 . An isolated nucleotide fragment comprising a nucleic acid sequence SEQ ID NO:2, encoding a cytochrome P450 polypeptide associated with controlling embryo/endosperm size during seed development having an amino acid identity of at least 65% based on the Clustal method of alignment when compared to SEQ ID NO: 97, or the complement thereof. 
     
     
         2 . The isolated nucleotide fragment of  claim 1 , wherein the polypeptide it encodes comprises a motif corresponding to the amino acid sequence set forth in SEQ ID NO:88 wherein said motif is a conserved subsequence. 
     
     
         3 . The isolated nucleotide fragment of  claim 1  or  2  wherein said fragment or part thereof is useful in antisense inhibition or co-suppression of a cytochrome P450 polypeptide associated with controlling embryo/endosperm size during seed development in a transformed plant. 
     
     
         4 . An isolated nucleic acid fragment comprising a promoter wherein said promoter consists essentially of the nucleotide sequence set forth in SEQ ID NOs:3, 4, 104, or 105, or said promoter consists essentially of a fragment or subfragment that is substantially similar and functionally equivalent to the nucleotide sequence set forth in SEQ ID NOs:3, 4, 104, or 105. 
     
     
         5 . A chimeric construct comprising the isolated nucleotide fragment of  claim 1  or  2  operably linked to at least one regulatory sequence. 
     
     
         6 . A chimeric construct comprising the isolated nucleic acid fragment of  claim 3  operably linked to at least one regulatory sequence. 
     
     
         7 . The chimeric construct of  claim 5  wherein said isolated nucleic acid fragment is operably linked to the promoter of  claim 4 . 
     
     
         8 . The chimeric construct of  claim 6  wherein said isolated nucleic acid fragment is operably linked to the promoter of  claim 4 . 
     
     
         9 . A plant comprising in its genome the chimeric construct of  claim 5 . 
     
     
         10 . A plant comprising in its genome the chimeric construct of  claim 6 . 
     
     
         11 . A plant comprising in its genome the chimeric construct of  claim 7 . 
     
     
         12 . A plant comprising in its genome the chimeric construct of  claim 8 . 
     
     
         13 . Seeds obtained from the plant of  claim 9 . 
     
     
         14 . Seeds obtained from the plant of  claim 10 . 
     
     
         15 . Seeds obtained from the plant of  claim 11 . 
     
     
         16 . Seeds obtained from the plant of  claim 12 . 
     
     
         17 . Oil obtained from the seeds of  claim 13 . 
     
     
         18 . Oil obtained from the seeds of  claim 14 . 
     
     
         19 . Oil obtained from the seeds of  claims 15 . 
     
     
         20 . Oil obtained from the seeds of  claim 16 . 
     
     
         21 . The plant of  claim 9  wherein said plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         22 . The plant of  claim 10  wherein said plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         23 . The plant of  claim 11  wherein said plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         24 . The plant of  claim 12  wherein said plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         25 . Transformed plant tissue or plant cells comprising the chimeric construct of  claim 5 . 
     
     
         26 . Transformed plant tissue or plant cells conprising the chimeric construct of  claim 6 . 
     
     
         27 . Transformed plant tissue or plant cells comprising the chimeric construct of  claim 7 . 
     
     
         28 . Transformed plant tissue or plant cells comprising the chimeric construct of  claim 8 . 
     
     
         29 . The plant tissue or plant cells of  claim 25  wherein the plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         30 . The plant tissue or plant cells of  claim 26  wherein the plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         31 . The plant tissue or plant cells of  claim 27  wherein the plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         32 . The plant tissue or plant cells of  claim 28  wherein the plant is selected from the group consisting of rice, corn, sorghum, millet, rye, soybean, canola, wheat, barley, oat, beans, and nuts. 
     
     
         33 . A method of controlling embryo/endosperm size during seed development in plants which comprises:
 (a) transforming a plant with the chimeric construct of  claim 5 ;   (b) growing the transformed plant under conditions suitable for the expression of the chimeric construct; and   (c) selecting those transformed plants which produce seeds having an altered embryo/endosperm size.   
     
     
         34 . A method of controlling embryo/endosperm size during seed development in plants which comprises:
 (a) transforming a plant with the chimeric construct of  claim 6 ;   (b) growing the transformed plant under conditions suitable for the expression of the chimeric construct; and   (c) selecting those transformed plants which produce seeds having an altered embryo/endosperm size.   
     
     
         35 . A method of controlling embryo/endosperm size during seed development in plants which comprises:
 (a) transforming a plant with the chimeric construct of  claim 7 ;   (b) growing the transformed plant under conditions suitable for the expression of the chimeric construct; and   (c) selecting those transformed plants which produce seeds having an altered embryo/endosperm size.   
     
     
         36 . A method of controlling embryo/endosperm size during seed development in plants which comprises:
 (a) transforming a plant with the chimeric construct of  claim 8 ;   (b) growing the transformed plant under conditions suitable for the expression of the chimeric construct; and   (c) selecting those transformed plants which produce seeds having an altered embryo/endosperm size.   
     
     
         37 . A method to isolate nucleic acid fragments encoding polypeptides associated with controlling embryo/endosperm size during seed development which comprises:
 (a) comparing SEQ ID NOs:2, 7, 9,11, 13.15,17,19. 21, 23, 25, 27, 29, 31, 33, 35, 37, 39,41,42, 43, 44, 45, 46, 47, 93, 95, 97, or 99 with other polypeptide sequences associated with controlling embryo/endosperm size during seed development;   (b) identifying the conserved sequences(s) or 4 or more amino acids obtained in step (a);   (c) making region-specific nucleotide probe(s) or oligomer(s) based on the conserved sequences identified in step (b); and   (d) using the nucleotide probe(s) or oligomer(s) of step (c) to isolate sequences associated with controlling embryo/endosperm size during seed development by sequence dependent protocols.   
     
     
         38 . A method of mapping genetic variations related to controlling embryo/endosperm size and/or altering oil phenotype in plants comprising:
 (a) crossing two plant varieties; and   (b) evaluating genetic variations with respect to
 (i) a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, 3, 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 92, 94, 96, 98, 100, 102, 104or 105; or 
 (ii) a nucleic acid sequence encoding a polypeptide selected from the group consisting of SEQ ID NO:2, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 42, 43, 44, 45, 46, 47, 80- 91, 93, 95, 9in progeny plants resulting from the cross of step (a) wherein the evaluation is made using a method selected from the group consisting of: RFLP analysis, SNP analysis, and PCR-based analysis.    
   
     
     
         39 . A method of molecular breeding to control embryo/endosperm size and/or altering oil phenotype in plants comprising:
 (a) crossing two plant varieties; and   (b) evaluating genetic variations with respect to
 (i) a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, 3, 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28. 30, 32, 34, 36, 38, 40, 92, 94, 96, 98, 100, 102, 104, or 105; or 
 (ii) a nucleic acid sequence encoding a polypeptide selected from the group consisting of SEQ ID NO:2, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 42, 43, 44, 45, 46, 47, 80-91, 93, 95, 97, or 99; in progeny plants resulting from the cross of step (a) wherein the evaluation is made using a method selected from the group consisting of: RFLP analysis, SNP analysis, and PCR-based analysis.