Compositions and methods to control insect pests

Patent metadata

Field	Value
Publication number	US-2017290338-A1
Application number	US-201715597295-A
Country	US
Kind code	A1
Filing date	May 17, 2017
Priority date	Aug 28, 2009
Publication date	Oct 12, 2017
Grant date	—

Abstract

Methods and compositions are provided which employ a silencing element that, when ingested by a pest, such as a Coleopteran plant pest or a Diabrotica plant pest, decrease the expression of a target sequence in the pest. In specific embodiments, the decrease in expression of the target sequence controls the pest and thereby the methods and compositions are capable of limiting damage to a plant. The present invention provides various target polynucleotides set forth in any one of SEQ ID NOS: 1-236 or active variants and fragments thereof, wherein a decrease in expression of one or more the sequences in the target pest controls the pest (i.e., has insecticidal activity). Further provided are silencing elements which when ingested by the pest decrease the level of the target polypeptide and thereby control the pest. In specific embodiment, the pest is D. virgifera virgifera, D. barberi, D. speciosa , or D. undecimpunctata howardi . Plants, plant part, bacteria and other host cells comprising the silencing elements or an active variant or fragment thereof of the invention are also provided.

First claim

That which is claimed:

1 . An isolated polynucleotide comprising a nucleotide sequence comprising:
(a) the nucleotide sequence comprising any one of nucleotides 1-380 of SEQ ID NO: 45; SEQ ID NO: 45; SEQ ID NO: 13; SEQ ID NO: 15; nucleotides 1-266 of SEQ ID NO:50; SEQ ID NO:50; SEQ ID NO: 18; SEQ ID NO: 29; nucleotides 1-675 of SEQ ID NO:37, SEQ ID NO: 37; SEQ ID NO:16; SEQ ID NO:32; SEQ ID NO:25; nucleotides 1-132 of SEQ ID NO: 40; or any one of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 14, 17, 19, 20, 21, 22, 23, 24, 26, 27, 28, 30, 31, 33, 34, 35, 36, 38, 39, 41, 42, 43, 44, 46, 47, 48, 49, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, or a complement thereof; (b) the nucleotide sequence comprising at least 90% sequence identity to any one of nucleotides 1-380 of SEQ ID NO: 45; SEQ ID NO: 45; SEQ ID NO: 13; SEQ ID NO: 15; nucleotides 1-266 of SEQ ID NO:50; SEQ ID NO:50; SEQ ID NO: 18; SEQ ID NO: 29; nucleotides 1-675 of SEQ ID NO:37, SEQ ID NO: 37; SEQ ID NO:16; SEQ ID NO:32; SEQ ID NO:25; nucleotides 1-132 of SEQ ID NO: 40; or any one of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 14, 17, 19, 20, 21, 22, 23, 24, 26, 27, 28, 30, 31, 33, 34, 35, 36, 38, 39, 41, 42, 43, 44, 46, 47, 48, 49, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, or a complement thereof; wherein said polynucleotide encodes a silencing element having insecticidal activity against a Coleopteran plant pest; or (c) the nucleotide sequence comprising at least 19 consecutive nucleotides of any one of nucleotides 1-380 of SEQ ID NO: 45; SEQ ID NO: 45; SEQ ID NO: 13; SEQ ID NO: 15; nucleotides 1-266 of SEQ ID NO:50; SEQ ID NO:50; SEQ ID NO: 18; SEQ ID NO: 29; nucleotides 1-675 of SEQ ID NO:37, SEQ ID NO: 37; SEQ ID NO:16; SEQ ID NO:32; SEQ ID NO:25; nucleotides 1-132 of SEQ ID NO: 40; or any one of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 14, 17, 19, 20, 21, 22, 23, 24, 26, 27, 28, 30, 31, 33, 34, 35, 36, 38, 39, 41, 42, 43, 44, 46, 47, 48, 49, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, or a complement thereof; wherein said polynucleotide encodes a silencing element having insecticidal activity against a Coleopteran plant pest.

2 . The isolated polynucleotide of claim 1 , wherein said Coleopteran plant pest is a Diabrotica plant pest.

3 . An expression cassette comprising the polynucleotide of claim 1 .

4 . The expression cassette of claim 3 , wherein said polynucleotide is operably linked to a heterologous promoter.

5 . The expression cassette of claim 3 , wherein said polynucleotide is expressed as a double stranded RNA.

6 . The expression cassette of claim 3 , wherein said polynucleotide comprise a silencing element which is expressed as a hairpin RNA.

7 . The expression cassette of claim 6 , wherein the silencing element comprises, in the following order, a first segment, a second segment, and a third segment, wherein
a) said first segment comprises at least about 19 nucleotides having at least 90% sequence complementarity to a target sequence set forth in any one of nucleotides 1-380 of SEQ ID NO: 45; SEQ ID NO: 45; SEQ ID NO: 13; SEQ ID NO: 15; nucleotides 1-266 of SEQ ID NO:50; SEQ ID NO:50; SEQ ID NO: 18; SEQ ID NO: 29; nucleotides 1-675 of SEQ ID NO:37, SEQ ID NO: 37; SEQ ID NO:16; SEQ ID NO:32; SEQ ID NO:25; nucleotides 1-132 of SEQ ID NO: 40; or any one of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 14, 17, 19, 20, 21, 22, 23, 24, 26, 27, 28, 30, 31, 33, 34, 35, 36, 38, 39, 41, 42, 43, 44, 46, 47, 48, 49, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, or 236; b) said second segment comprises a loop of sufficient length to allow the silencing element to be transcribed as a hairpin RNA; and, c) said third segment comprises at least about 19 nucleotides having at least 85% complementarity to the first segment.

8 . The expression cassette of claim 3 , wherein said 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 polynucleotide.

9 . A host cell comprising a heterologous expression cassette of claim 3 .

10 . A plant cell having stably incorporated into its genome a heterologous polynucleotide comprising a silencing element, wherein said silencing element comprises
a) a fragment of at least 19 consecutive nu

Assignees

• Du Pont

Inventors

• Broglie Karen E
• Kriss Kevin
• Lu Albert
• Muthalagi Mani
• Presnail James Kevin

Classifications

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• C12N2310/531

  Stem-loop; Hairpin · CPC title

• C12N2310/11

  Antisense · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• A01N65/20

  Fabaceae or Leguminosae [Pea or Legume family], e.g. pea, lentil, soybean, clover, acacia, honey locust, derris or millettia · CPC title