Compositions and methods to control insect pests

Abstract

Methods and compositions are provided which employ a silencing element that, when ingested by a plant insect pest, such as Coleopteran, Hemiptera, or Lepidopteran plant pest, including a Diabrotica, Leptinotarsa, Phyllotreta, Acyrthosiphan, Bemisia, Halyomorpha, Nezara, or Spodoptera plant pest, decrease the expression of a target sequence in the pest. Disclosed are various target polynucleotides set forth in any one of SEQ ID NOS: 1-53 or 107-254 disclosed herein, or variants and fragments thereof, or complements thereof, wherein a decrease in expression of one or more of the sequences in the target pest controls the pest and pest population by insect sterilization (i.e., male and female sterility, reduction of sperm count, egg production and viability) and mating-based sterile insect technique (SIT). Plants, plant parts, bacteria and other host cells comprising the silencing elements or an active variant or fragment thereof of the invention are also provided. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.

1 . A silencing element comprising at least one double-stranded RNA region, at least one strand of which comprises a polynucleotide that is complementary to:
 (a) the nucleotide sequence comprising any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof;   (b) the nucleotide sequence comprising at least 90% sequence identity to any one of nucleotides SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof; or   (c) the nucleotide sequence comprising at least 19 consecutive nucleotides of any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof;   wherein the silencing element has sterilization activity against an insect plant pest.   
     
     
         2 . (canceled) 
     
     
         3 . The silencing element of  claim 1 , wherein the insect plant pest is a Coleoptera plant pest. 
     
     
         4 . The silencing element of  claim 3 , wherein the Coleoptera plant pest is a  Diabrotica  plant pest. 
     
     
         5 . The silencing element of  claim 4 , wherein the  Diabrotica  plant pest comprises  D. virgifera virgifera, D. virgifera zeae, D. speciosa, D. barberi, D. virgifera zeae , or  D. undecimpunctata howardi.    
     
     
         6 - 19 . (canceled) 
     
     
         20 . The silencing element of  claim 1 , wherein the silencing element comprises a hairpin loop. 
     
     
         21 . The silencing element of  claim 20 , wherein the silencing element comprises, a first segment, a second segment, and a third segment, wherein
 a. the first segment comprises at least about 19 nucleotides having at least 90% sequence complementarity to a sequence set forth in any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments, and complements thereof; or the first segment consists of at least 19 nucleotides having at least 90% sequence complementarity to a sequence set forth in any one of SEQ ID NOS: 1-53 or 107-253;   b. the second segment comprises a loop of sufficient length to allow the silencing element to be transcribed as a hairpin RNA; and,   c. the third segment comprises at least about 19 nucleotides having at least 85% complementarity to the first segment;   wherein the second segment that is not complementary a sequence set forth in any one of SEQ ID NOS: 1-53 or 107-253, or to the other segments of the construct; and   wherein the first and third segments form at least a partially double-stranded region.   
     
     
         22 - 25 . (canceled) 
     
     
         26 . The silencing element of  claim 21 , wherein the first segment is complementary to a Coleoptera insect species; and wherein the third segment is complementary to a different Coleoptera insect species. 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . A DNA construct comprising a polynucleotide encoding the silencing element of  claim 1 . 
     
     
         30 . An expression construct comprising a DNA construct of  claim 29 . 
     
     
         31 . The expression cassette of  claim 30 , wherein the polynucleotide is operably linked to a heterologous promoter. 
     
     
         32 . The expression cassette of  claim 30 , wherein the polynucleotide is flanked by a first operably linked convergent promoter at one terminus of the polynucleotide and a second operably linked convergent promoter at the opposing terminus of the polynucleotide, wherein the first and the second convergent promoters are capable of driving expression of the silencing element. 
     
     
         33 . A host cell comprising the DNA construct of  claim 29 . 
     
     
         34 . The host cell of  claim 33 , wherein the host cell is a bacterial cell. 
     
     
         35 . The host cell of  claim 34 , wherein the bacterial cell is an inactivated bacterial cell. 
     
     
         36 . A host cell comprising the expression construct of  claim 30 . 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . A composition comprising the ribonucleic acid construct of  claim 1 . 
     
     
         40 . The composition of  claim 39 , further comprising an agriculturally acceptable carrier. 
     
     
         41 . The composition of  claim 39 , further comprising a herbicide compound, an insecticide, a fungicide, a nematocide, an agriculturally-acceptable carrier, and/or a bacteria, or combinations thereof. 
     
     
         42 . The composition of  claim 39 , wherein the composition is in liquid form, solid form, or gel form. 
     
     
         43 . (canceled) 
     
     
         44 . The composition of  claim 42 , wherein the solid form is a pellet, a powder, an aggregate, or a molded article. 
     
     
         45 . A plant cell having stably incorporated into its genome a heterologous polynucleotide encoding a silencing element, wherein the polynucleotide comprises:
 a. the nucleotide sequence comprising any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof;   b. the nucleotide sequence comprising at least 90% sequence identity to any one of nucleotides SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof; or   c. the nucleotide sequence comprising at least 19 consecutive nucleotides of any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof;   wherein the silencing element decreases the fertility of an insect plant pest.   
     
     
         46 . (canceled) 
     
     
         47 . The plant cell of  claim 45 , wherein the insect plant pest is a Coleoptera plant pest. 
     
     
         48 . The plant cell of  claim 47 , wherein the Coleoptera plant pest is a  Diabrotica  plant pest. 
     
     
         49 . The plant cell of  claim 48 , wherein the  Diabrotica  plant pest comprises  D. virgifera virgifera, D. virgifera zeae, D. speciosa, D. barberi, D. virgifera zeae , or  D. undecimpunctata howardi.    
     
     
         50 .- 63 . (canceled) 
     
     
         64 . The plant cell of  claim 45 , wherein the plant cell further comprises an expression cassette, wherein the expression cassette comprises the heterologous polynucleotide encoding a silencing element. 
     
     
         65 . The plant cell of  claim 45 , wherein the silencing element expresses a double stranded RNA. 
     
     
         66 . The plant cell of  claim 45 , wherein the silencing element expresses a hairpin RNA. 
     
     
         67 . The plant cell of  claim 45 , wherein the polynucleotide is operably linked to a heterologous promoter. 
     
     
         68 . The plant cell of  claim 45 , wherein the plant cell is from a monocot. 
     
     
         69 . The plant cell of  claim 68 , wherein the monocot is maize, barley, millet, wheat or rice. 
     
     
         70 . The plant cell of  claim 45 , wherein the plant cell is from a dicot. 
     
     
         71 . The plant cell of  claim 70 , wherein the dicot is kale, cauliflower, broccoli, mustard plant, cabbage, pea, clover, alfalfa, broad bean, tomato, cassava, soybean, canola, alfalfa, sunflower, safflower, tobacco,  Arabidopsis , or cotton. 
     
     
         72 . A plant or plant part comprising the plant cell of  claim 45 . 
     
     
         73 . A transgenic seed from the plant of  claim 72 . 
     
     
         74 . A method for controlling a plant insect pest comprising feeding to a plant insect pest a composition comprising a silencing element, wherein the silencing element controls the plant pest, wherein the silencing element comprises a sequence complementary to:
 a. the nucleotide sequence comprising any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof;   b. the nucleotide sequence comprising at least 90% sequence identity to any one of nucleotides SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof; or   c. the nucleotide sequence comprising at least 19 consecutive nucleotides of any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments thereof, and complements thereof; or   wherein the silencing element has insect sterilization activity against the plant pest.   
     
     
         75 . The method of  claim 74 , wherein the composition comprises a plant or plant part having stably incorporated into its genome a polynucleotide encoding the silencing element. 
     
     
         76 . The method of  claim 74 , wherein the silencing element comprises a double stranded RNA. 
     
     
         77 . The method of  claim 74 , wherein the silencing element comprises a hairpin RNA. 
     
     
         78 . The method of  claim 75 , wherein the polynucleotide encoding silencing element is operably linked to a heterologous promoter. 
     
     
         79 . (canceled) 
     
     
         80 . The method of  claim 74 , wherein the silencing element comprises, a first segment, a second segment, and a third segment, wherein
 a. the first segment comprises at least about 19 nucleotides having at least 90% sequence complementarity to a sequence set forth in any one of SEQ ID NOS: 1-53 or 107-253; or variants and fragments, and complements thereof,   b. the second segment comprises a loop of sufficient length to allow the silencing element to be transcribed as a hairpin RNA; and,   c. the third segment comprises at least about 19 nucleotides having at least 85% complementarity to the first segment;   wherein the second segment that is not complementary a sequence set forth in any one of SEQ ID NOS: 1-53 or 107-253; or to the other segments of the construct; and   wherein the first and third segments form at least a partially double-stranded region.   
     
     
         81 - 84 . (canceled) 
     
     
         85 . The method of  claim 80 , wherein the first segment is complementary to a Coleoptera insect species; and wherein the third segment is complementary to a different Coleoptera insect species. 
     
     
         86 . (canceled) 
     
     
         87 . (canceled) 
     
     
         88 . The method of  claim 74 , wherein the plant is a monocot. 
     
     
         89 . The method of  claim 88 , wherein the monocot is maize, barley, millet, wheat or rice. 
     
     
         90 . The method of  claim 74 , wherein the plant is a dicot. 
     
     
         91 . The method of  claim 90 , wherein the dicot is kale, cauliflower, broccoli, mustard plant, cabbage, pea, clover, alfalfa, broad bean, tomato, cassava, soybean, canola, alfalfa, sunflower, safflower, tobacco,  Arabidopsis , or cotton. 
     
     
         92 - 110 . (canceled)