Parental rnai suppression of hunchback gene to control hemipteran pests

Abstract

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

1 . A method for controlling a hemipteran pest population, the method comprising providing an agent comprising a ribonucleic acid (RNA) molecule that functions upon contact with the hemipteran pest to inhibit a biological function within the hemipteran pest, wherein the RNA is specifically hybridizable with a polynucleotide selected from the group consisting of SEQ ID NOs:12 and 13; the complement of either of SEQ ID NOs:12 and 13; a fragment of at least 15 contiguous nucleotides of either of SEQ ID NOs:12 and 13; the complement of a fragment of at least 15 contiguous nucleotides of either of SEQ ID NOs:12 and 13; a transcript of either of SEQ ID NO:2; the complement of a transcript of either of SEQ ID NO:2; a fragment of at least 15 contiguous nucleotides of a transcript of either of SEQ ID NO:2; and the complement of a fragment of at least 15 contiguous nucleotides of a transcript of either of SEQ ID NO:2. 
     
     
         2 . The method according to  claim 1 , wherein the agent is a double-stranded RNA molecule. 
     
     
         3 . A method for controlling a hemipteran pest population, the method comprising:
 introducing into a hemipteran pest, a ribonucleic acid (RNA) molecule that functions upon contact with the hemipteran pest to inhibit a biological function within the hemipteran pest, wherein the RNA is specifically hybridizable with a polynucleotide selected from the group consisting of any of
 SEQ ID NOs:12 and 13, 
 the complement of either of SEQ ID NOs:12 and 13, 
 a fragment of at least 15 contiguous nucleotides of either of SEQ ID NOs:12 and 13, 
 the complement of a fragment of at least 15 contiguous nucleotides of either of SEQ ID NOs:12 and 13, 
 a transcript of either of SEQ ID NO:2, 
 the complement of a transcript of either of SEQ ID NO:2, 
 a fragment of at least 15 contiguous nucleotides of a transcript of either of SEQ ID NO:2, and 
 the complement of a fragment of at least 15 contiguous nucleotides of a transcript of either of SEQ ID NO:2, 
 thereby producing a hemipteran pest having a pRNAi phenotype. 
   
     
     
         4 . The method according to  claim 3 , wherein the RNA is introduced into a male hemipteran pest. 
     
     
         5 . The method according to  claim 3 , wherein the RNA is introduced into a female hemipteran pest, the method further comprising releasing the female hemipteran pest having the pRNAi phenotype into the pest population, wherein mating between the female hemipteran pest having the pRNAi phenotype and male pests of the population produces fewer viable offspring than mating between other female pests and male pests of the population. 
     
     
         6 . A method for controlling a hemipteran pest population, the method comprising:
 providing an agent comprising a first and a second polynucleotide sequence that functions upon contact with the hemipteran pest to inhibit a biological function within the hemipteran pest, wherein the first polynucleotide sequence comprises a region that exhibits from about 90% to about 100% sequence identity to from about 19 to about 30 contiguous nucleotides of SEQ ID NO:12, and wherein the first polynucleotide sequence is specifically hybridized to the second polynucleotide sequence.   
     
     
         7 . The method according to  claim 6 , wherein the ribonucleic acid molecule is a double-stranded ribonucleic acid molecule. 
     
     
         8 . The method according to  claim 6 , wherein the hemipteran pest population is reduced relative to a population of the same pest species infesting a host plant of the same host plant species lacking the transformed plant cell. 
     
     
         9 . A method for controlling a hemipteran pest population, the method comprising:
 providing in a host plant of a hemipteran pest a transformed plant cell comprising the polynucleotide of  claim 1 , wherein the polynucleotide is expressed to produce a ribonucleic acid molecule that functions upon contact with a hemipteran pest belonging to the population to inhibit the expression of a target sequence within the hemipteran pest and results in decreased reproduction of the hemipteran pest or pest population, relative to reproduction of the same pest species on a plant of the same host plant species that does not comprise the polynucleotide.   
     
     
         10 . The method according to  claim 9 , wherein the ribonucleic acid molecule is a double-stranded ribonucleic acid molecule. 
     
     
         11 . The method according to  claim 9 , wherein the hemipteran pest population is reduced relative to a hemipteran pest population infesting a host plant of the same species lacking the transformed plant cell. 
     
     
         12 . A method of controlling hemipteran pest infestation in a plant, the method comprising providing in the diet of a hemipteran pest a ribonucleic acid (RNA) that is specifically hybridizable with a polynucleotide selected from the group consisting of:
 SEQ ID NOs:12 and 13;   the complement of either of SEQ ID NOs:12 and 13;   a fragment of at least 15 contiguous nucleotides of either of SEQ ID NOs:12 and 13;   the complement of a fragment of at least 15 contiguous nucleotides of either of SEQ ID NOs:12 and 13;   a transcript of any of either of SEQ ID NOs:1 and 2;   the complement of a transcript of either of SEQ ID NOs:1 and 2;   a fragment of at least 15 contiguous nucleotides of a transcript of either of SEQ ID NOs:1 and 2; and   the complement of a fragment of at least 15 contiguous nucleotides of a transcript of either of SEQ ID NOs:1 and 2.   
     
     
         13 . The method according to  claim 12 , wherein the diet comprises a plant cell transformed to express the polynucleotide. 
     
     
         14 . The method according to  claim 12 , wherein the specifically hybridizable RNA is comprised in a double-stranded RNA molecule. 
     
     
         15 . A method for improving the yield of a corn crop, the method comprising:
 introducing the nucleic acid of  claim 1  into a corn plant to produce a transgenic corn plant; and   cultivating the corn plant to allow the expression of the at least one polynucleotide; wherein expression of the at least one polynucleotide inhibits hemipteran pest reproduction or growth and loss of yield due to hemipteran pest infection.   
     
     
         16 . The method according to  claim 15 , wherein expression of the at least one polynucleotide produces an RNA molecule that suppresses at least a first target gene in a hemipteran pest that has contacted a portion of the plant. 
     
     
         17 . A method for producing a transgenic plant cell, the method comprising:
 transforming a plant cell with a vector comprising the nucleic acid of  claim 1 ;   culturing the transformed plant cell under conditions sufficient to allow for development of a plant cell culture comprising a plurality of transformed plant cells;   selecting for transformed plant cells that have integrated the at least one polynucleotide into their genomes;   screening the transformed plant cells for expression of a ribonucleic acid (RNA) molecule encoded by the at least one polynucleotide; and   selecting a plant cell that expresses the RNA.   
     
     
         18 . The method according to  claim 17 , wherein the RNA molecule is a double-stranded RNA molecule. 
     
     
         19 . A method for producing a hemipteran pest-resistant transgenic plant, the method comprising:
 providing the transgenic plant cell produced by the method of  claim 17 ; and   regenerating a transgenic plant from the transgenic plant cell, wherein expression of the ribonucleic acid molecule encoded by the at least one polynucleotide is sufficient to modulate the expression of a target gene in a hemipteran pest that contacts the transformed plant.   
     
     
         20 . A method for producing a transgenic plant cell, the method comprising:
 transforming a plant cell with a vector comprising a means for protecting a plant from a hemipteran pest;   culturing the transformed plant cell under conditions sufficient to allow for development of a plant cell culture comprising a plurality of transformed plant cells;   selecting for transformed plant cells that have integrated the means for protecting a plant from a hemipteran pest into their genomes;   screening the transformed plant cells for expression of a means for inhibiting expression of an essential gene in a hemipteran pest; and   selecting a plant cell that expresses the means for inhibiting expression of an essential gene in a hemipteran pest.   
     
     
         21 . A method for producing a hemipteran pest-resistant transgenic plant, the method comprising:
 providing the transgenic plant cell produced by the method of  claim 20 ; and   regenerating a transgenic plant from the transgenic plant cell, wherein expression of the means for inhibiting expression of an essential gene in a hemipteran pest is sufficient to modulate the expression of a target gene in a hemipteran pest that contacts the transformed plant.   
     
     
         22 . The method according to  claim 21 , wherein the transformed plant cell comprises a nucleotide sequence encoding a polypeptide from  Bacillus thuringiensis, Alcaligenes  spp., or  Pseudomonas  spp. 
     
     
         23 . The method according to  claim 22 , wherein the polypeptide from  B. thuringiensis  is selected from a group comprising Cry1A, Cry2A, Cry3A, Cry11A, and Cry51A.