Materials and methods for the modulation of cyclin-dependent kinase inhibitor-like polypeptides in maize

Abstract

Isolated or purified nucleic acid molecules encoding polypeptides having cyclin-dependent kinase (CDK) inhibitor-like activity; vectors; host cells; polypeptides having CDK inhibitor-like activity; nucleic acid constructs for suppression of CDK inhibitor-like activity; methods of suppressing and up-regulating the expression of one or more CDK inhibitor-like genes in a maize cell, tissue, organ, or plant; a maize cell, tissue, organ, or plant in which the expression of a CDK inhibitor-like gene has been suppressed or up-regulated in accordance with such methods; and a seed (and the oil and meal thereof) obtained from a plant in which the expression of one or more CDK inhibitor-like genes has been suppressed or up-regulated.

1 - 7 . (canceled) 
     
     
         8 . A nucleic acid construct comprising at least one transcribable nucleotide sequence, the expression of which results in the suppression of an endogenous nucleic acid selected from the group consisting of: SEQ ID NO: 1 [ZmKRP1], a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 2 [ZmKRP1], SEQ ID NO: 3 [ZmKRP2], and a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 4 [ZmKRP2], alone or in further combination with at least one other nucleotide sequence selected from the group consisting of SEQ ID NO: 5 [ZmKRP3], a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 6 [ZmKRP3], SEQ ID NO: 7 [ZmKRP4], a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8 [ZmKRP4], SEQ ID NO: 9 [ZmKRP5], a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 10 [ZmKRP5], SEQ ID NO: 11 [ZmKRP6], a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 12 [ZmKRP6], SEQ ID NO: 13 [ZmKRP7], a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 14 [ZmKRP7], SEQ ID NO: 15 [ZmKRP8], a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 16 [ZmKRP8], SEQ ID NO: 17 [ZmKRP9], and a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 18 [ZmKRP9], wherein, when the nucleic acid construct comprises two or more transcribable nucleotide sequences, the nucleotide sequences can be present in any order on the nucleic acid construct and can be polycistronic, wherein each transcribable nucleotide sequence comprises at least about 100 contiguous nucleotides, and wherein the expression of each transcribable nucleotide sequence optionally induces gene suppression. 
     
     
         9 . The nucleic acid construct of  claim 8 , which further comprises a promoter that is preferentially expressed in the germ and/or aleurone of a kernel of maize and the promoter is operably linked to at least one transcribable nucleotide sequence. 
     
     
         10 . The nucleic acid construct of  claim 9 , wherein the promoter is an oleosin promoter of maize, a globulin promoter of maize, a MIP synthase promoter of maize, a Perl promoter of barley, an embryo-preferred promoter such as P-Os.CPC214 from rice or an embryo-preferred promoter such as P-Zm.CEP1, P-Zm.CPC214, P-Zm.CPC214tr1, or P-Zm.CPC214tr2 from maize. 
     
     
         11 . The nucleic acid construct of  claim 8 , wherein the at least one transcribable nucleotide sequence is a noncoding sequence. 
     
     
         12 . The nucleic acid construct of  claim 11 , wherein the noncoding sequence is an intron, a nucleotide sequence from a promoter region, a nucleotide sequence from a 5′ untranslated region, a nucleotide sequence from a 3′ untranslated region, or a fragment of any of the foregoing. 
     
     
         13 - 22 . (canceled) 
     
     
         23 . A method of suppressing the expression of one or more CDK inhibitor-like genes in a maize cell, a maize tissue, a maize organ, or a maize plant, which method comprises contacting said maize cell, maize tissue, maize organ or maize plant with a nucleic acid construct of  claims 8 ,  14 , or  18 . 
     
     
         24 . The method of  claim 23 , wherein the suppression is accomplished by antisense suppression, sense suppression, dsRNA, ribozyme, or a combination thereof. 
     
     
         25 . The method of  claim 23 , wherein the nucleic acid construct further comprises a promoter that is preferentially expressed in the germ and/or aleurone of a kernel of maize and the promoter is operably linked to the at least one nucleotide sequence. 
     
     
         26 . The method of  claim 25 , wherein the promoter is an oleosin promoter of maize, a globulin promoter of maize, a MIP synthase promoter of maize, a Perl promoter of barley, an embryo-preferred promoter such as P-Os.CPC214 from rice or an embryo-preferred promoter such as P-Zm.CEP1, P-Zm.CPC214, P-Zm.CPC214tr1, or P-Zm.CPC214tr2 from maize. 
     
     
         27 . A maize cell, a maize tissue, a maize organ, or a maize plant in which the expression of a CDK inhibitor-like gene has been suppressed in accordance with the method of  claim 23 . 
     
     
         28 . A seed obtained from a plant of  claim 27 . 
     
     
         29 - 30 . (canceled) 
     
     
         31 . The method of  claim 23  wherein the suppression results in increased oil.