Parental rnai suppression of chromatin remodeling genes to control coleopteran pests

Abstract

This disclosure concerns nucleic acid molecules and methods of use thereof for control of coleopteran pests through RNA interference-mediated inhibition of target coding and transcribed non-coding sequences in coleopteran pests. The disclosure also concerns methods for making transgenic plants that express nucleic acid molecules useful for the control of coleopteran pests, and the plant cells and plants obtained thereby.

1 . An isolated nucleic acid molecule comprising a polynucleotide operably linked to a heterologous promoter, wherein the polynucleotide comprises at least a first nucleotide sequence selected from the group consisting of:
 at least 15 contiguous nucleotides of SEQ ID NO:79; the complement of at least 15 contiguous nucleotides of SEQ ID NO:79, at least 15 contiguous nucleotides of a native coding sequence of a  Diabrotica  organism comprising SEQ ID NO:101, and the complement of at least 15 contiguous nucleotides of a native coding sequence of a  Diabrotica  organism comprising SEQ ID NO:101.   
     
     
         2 . The nucleic acid molecule of  claim 1 , wherein the first nucleotide sequence is selected from the group consisting of SEQ ID NO:79, the complement of SEQ ID NO:79, SEQ ID NO:164, and the complement of SEQ ID NO:164. 
     
     
         3 . The nucleic acid molecule of  claim 1 , wherein the first nucleotide sequence comprises SEQ ID NO:101 or the complement of SEQ ID NO:101. 
     
     
         4 . The nucleic acid molecule of  claim 1 , wherein the heterologous promoter is functional in a plant cell. 
     
     
         5 . The nucleic acid molecule of  claim 4 , wherein the molecule is a plant transformation vector. 
     
     
         6 . A double-stranded ribonucleic acid (dsRNA) molecule comprising a polyribonucleotide encoded by the polynucleotide of the nucleic acid molecule of  claim 1 . 
     
     
         7 . The dsRNA molecule of  claim 6 , wherein the molecule comprises the polyribonucleotide as a single hybridized strand in the molecule. 
     
     
         8 . The nucleic acid molecule of  claim 1 , wherein the polynucleotide encodes a double-stranded ribonucleic acid (dsRNA) molecule, the polynucleotide further comprising a second nucleotide sequence and a third nucleotide sequence, wherein the third nucleotide sequence is linked to the first nucleotide sequence by the second nucleotide sequence, and wherein the third nucleotide sequence encodes a ribonucleotide sequence that hybridizes to a ribonucleotide sequence encoded by the first nucleotide sequence in a stem structure in the dsRNA molecule. 
     
     
         9 . The nucleic acid molecule of  claim 8 , wherein the heterologous promoter is functional in a plant cell. 
     
     
         10 . A double-stranded ribonucleic acid (dsRNA) molecule comprising a first, a second, and a third ribonucleotide sequence,
 wherein the first ribonucleotide sequence is selected from the group consisting of at least 15 contiguous ribonucleotides of SEQ ID NO:143; the complement of at least 15 contiguous ribonucleotides of SEQ ID NO:143, at least 15 contiguous ribonucleotides of a native transcript of a  Diabrotica  organism comprising SEQ ID NO:158, and the complement of at least 15 contiguous ribonucleotides of a native transcript of a  Diabrotica  organism comprising SEQ ID NO:158,   wherein the third ribonucleotide sequence is linked to the first ribonucleotide sequence by the second ribonucleotide sequence, and   wherein the first and the third ribonucleotide sequences hybridize in the stem of a hairpin structure in the dsRNA molecule.   
     
     
         11 . The dsRNA molecule of  claim 10 , wherein the first ribonucleotide sequence is selected from the group consisting of SEQ ID NO:143, the complement of SEQ ID NO:143, SEQ ID NO:167, and the complement of SEQ ID NO:167. 
     
     
         12 . The dsRNA molecule of  claim 10 , wherein the first ribonucleotide sequence comprises SEQ ID NO:158 or the complement of SEQ ID NO:158. 
     
     
         13 . A prokaryotic cell comprising the nucleic acid molecule of  claim 1 . 
     
     
         14 . A eukaryotic cell comprising the nucleic acid molecule of  claim 1 . 
     
     
         15 . A transgenic plant material comprising the nucleic acid molecule of  claim 9 . 
     
     
         16 . The transgenic plant material of  claim 15 , wherein the plant material is a plant cell. 
     
     
         17 . The transgenic plant material of  claim 15 , wherein the plant material is a plant part, plant organ, plant tissue, callus, callus culture, or plant tissue culture, and wherein the heterologous promoter is functional in the plant material. 
     
     
         18 . The transgenic plant material of  claim 15 , wherein the plant material is from  Zea mays.    
     
     
         19 . The transgenic plant material of  claim 15 , wherein the plant material is a seed or viable plant. 
     
     
         20 . The transgenic seed or viable plant of  claim 19 , wherein the seed or plant is  Zea mays.    
     
     
         21 . A transgenic plant commodity product comprising a detectable amount of the nucleic acid molecule of  claim 9 . 
     
     
         22 . A transgenic plant commodity product comprising a detectable amount of a double-stranded ribonucleic acid (dsRNA) molecule comprising a polyribonucleotide that comprises a ribonucleotide sequence selected from the group consisting of:
 at least 15 contiguous ribonucleotides of SEQ ID NO:143; the complement of at least 15 contiguous ribonucleotides of SEQ ID NO:143, at least 15 contiguous ribonucleotides of a native transcript of a  Diabrotica  organism comprising SEQ ID NO:158, and the complement of at least 15 contiguous nucleotides of a native transcript of a  Diabrotica  organism comprising SEQ ID NO:158.   
     
     
         23 . A method for controlling a coleopteran insect population, the method comprising feeding an insect of the population with an agent comprising the dsRNA molecule of  claim 6 . 
     
     
         24 . The method according to  claim 23 , wherein the agent is a sprayable formulation. 
     
     
         25 . A method for controlling a coleopteran insect population, the method comprising feeding an insect of the population with an agent comprising the dsRNA molecule of  claim 10 . 
     
     
         26 . The method according to  claim 25 , wherein the agent is a sprayable formulation. 
     
     
         27 . The method according to  claim 25 , wherein the agent is a transgenic host plant of a coleopteran insect or a part thereof, comprising a polynucleotide that is expressed in the plant to produce the dsRNA molecule. 
     
     
         28 . A method for improving the yield of a corn crop, the method comprising cultivating in the corn crop the transgenic seed or plant of  claim 19 . 
     
     
         29 . The method according to  claim 28 , wherein the transgenic seed or plant is a corn seed or plant. 
     
     
         30 . A method for producing a transgenic plant cell, the method comprising:
 transforming a plant cell with the nucleic acid molecule of  claim 9 ;   culturing the transformed plant cell under conditions sufficient to allow for development of a plant cell culture comprising a plurality of plant cells comprising the polynucleotide; and   selecting a transgenic plant cell that has integrated the polynucleotide into its genome, and that expresses the dsRNA molecule.   
     
     
         31 . A method for producing a transgenic plant, the method comprising regenerating a transgenic plant from the transgenic plant material of  claim 17 . 
     
     
         32 . The nucleic acid molecule of  claim 9 , further comprising a polynucleotide encoding a bacterial insecticidal polypeptide. 
     
     
         33 . The nucleic acid molecule of  claim 32 , wherein the bacterial insecticidal polypeptide is from  Bacillus thuringiensis,  and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C. 
     
     
         34 . The transgenic plant material of  claim 15 , wherein the plant material comprises a bacterial insecticidal polypeptide. 
     
     
         35 . The transgenic plant material of  claim 34 , wherein the bacterial insecticidal polypeptide is from  B. thuringiensis,  and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C. 
     
     
         36 . The transgenic seed or plant of  claim 19 , wherein the seed or plant comprises a bacterial insecticidal polypeptide. 
     
     
         37 . The transgenic seed or plant of  claim 36 , wherein the bacterial insecticidal polypeptide is from  B. thuringiensis,  and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C. 
     
     
         38 . The method according to  claim 23 , wherein the method further comprises feeding the insect of the population with a bacterial insecticidal polypeptide. 
     
     
         39 . The method according to  claim 38 , wherein the bacterial insecticidal polypeptide is from  B. thuringiensis,  and is selected from the group consisting of Cry1B, Cry3, Cry34, Cry35, Cry1I, Cry2A, Cry3, Cry7A, Cry8, Cry9D, Cry14, Cry18, Cry22, Cry23, Cry34, Cry35, Cry36, Cry37, Cry43, Cry55, Cyt1A, and Cyt2C.