Rna polymerase ii33 nucleic acid molecules to control insect pests

Patent metadata

Field	Value
Publication number	US-2016355841-A1
Application number	US-201615069689-A
Country	US
Kind code	A1
Filing date	Mar 14, 2016
Priority date	Mar 13, 2015
Publication date	Dec 8, 2016
Grant date	—

Abstract

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

First claim

What may be claimed is:

1 . An isolated nucleic acid comprising at least one polynucleotide operably linked to a heterologous promoter, wherein the polynucleotide is selected from the group consisting of:
SEQ ID NO:1; the complement of SEQ ID NO:1; a fragment of at least 15 contiguous nucleotides of SEQ ID NO:1; the complement of a fragment of at least 15 contiguous nucleotides of SEQ ID NO:1; a native coding sequence of a Diabrotica organism comprising SEQ ID NO:5; the complement of a native coding sequence of a Diabrotica organism comprising SEQ ID NO:5; a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Diabrotica organism comprising SEQ ID NO:5; the complement of a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Diabrotica organism comprising SEQ ID NO:5; SEQ ID NO:3; the complement of SEQ ID NO:3; a fragment of at least 15 contiguous nucleotides of SEQ ID NO:3; the complement of a fragment of at least 15 contiguous nucleotides of SEQ ID NO:3; a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:6-8; the complement of a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:6-8; a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:6-8; the complement of a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:6-8; SEQ ID NO:76; the complement of SEQ ID NO:76; a fragment of at least 15 contiguous nucleotides of SEQ ID NO:76; the complement of a fragment of at least 15 contiguous nucleotides of SEQ ID NO:76; a native coding sequence of a Euschistus organism comprising SEQ ID NO:80 or SEQ ID NO:81; the complement of a native coding sequence of a Euschistus organism comprising SEQ ID NO:80 or SEQ ID NO:81; a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Euschistus organism comprising SEQ ID NO:80 or SEQ ID NO:81; the complement of a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Euschistus organism comprising SEQ ID NO:80 or SEQ ID NO:81; SEQ ID NO:78; the complement of SEQ ID NO:78; a fragment of at least 15 contiguous nucleotides of SEQ ID NO:78; the complement of a fragment of at least 15 contiguous nucleotides of SEQ ID NO:78; a native coding sequence of a Euschistus organism comprising SEQ ID NO:82; the complement of a native coding sequence of a Euschistus organism comprising SEQ ID NO:82; a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Euschistus organism comprising SEQ ID NO:82; and the complement of a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Euschistus organism comprising SEQ ID NO:82.

2 . The polynucleotide of claim 1 , wherein the polynucleotide is selected from the group consisting of SEQ ID NO:1; the complement of SEQ ID NO:1; SEQ ID NO:3; the complement of SEQ ID NO:3; a fragment of at least 15 contiguous nucleotides of SEQ ID NO:1; the complement of a fragment of at least 15 contiguous nucleotides of SEQ ID NO:1; a fragment of at least 15 contiguous nucleotides of SEQ ID NO:3; the complement of a fragment of at least 15 contiguous nucleotides of SEQ ID NO:3; a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:5-8; the complement of a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:5-8; a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:5-8; and the complement of a fragment of at least 15 contiguous nucleotides of a native coding sequence of a Diabrotica organism comprising any of SEQ ID NOs:5-8.

3 . The polynucleotide of claim 1 , wherein the polynucleotide is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and the complements of any of the foregoing.

4 . The polynucleotide of claim 3 , wherein the organism is selected from the group consisting of D. v. virgifera LeConte; D. barberi Smith and Lawrence; D. u. howardi; D. v. zeae; D. balteata LeConte; D. u. tenella; D. speciosa Germar; and D. u. undecimpunctata Mannerheim.

5 . A plant transformation vector comprising the polynucleotide of claim 1 .

6 . A ribonucleic acid (RNA) molecule transcribed from the polynucleotide of claim 1 .

7 . A double-stranded RNA molecule produced from the expression of the polynucleotide of claim 1 .

8 . The double-stranded ribonucleic acid molecule of claim 7 , wherein contacting the polynucleotide sequence with a coleopteran or hemipteran insect inhibits the expression of an endogenous nucleotide sequence specifically complementary to the polynucleotide.

9 . The double-stranded ribonucleic acid molecule of claim 8 , wherein contacting said ribonucleotide molecule with a coleopteran or hemipteran insect kills or inhibits the growth, reproduction, and/or feeding of the insect.

10 . The double stranded RNA of claim 7 , comprising a first, a second and a third RNA segment, wherein the first RNA segment comprises the polynucleotide, wherein the third RNA segment is linked to the first RNA segment by the second polynucleotide sequence, and wherein the third RNA segment is substantially the reverse complement of the first RNA segment, such that the first and the third RNA segments hybridize when transcribed into a ribonucleic acid to form the double-stranded RNA.

11 . The RNA of claim 6 , selected from the group consisting of a double-stranded ribonucleic acid molecule and a single-stranded ribonucleic acid molecule of between about 15 and about 30 nucleotides in length.

12 . A plant transformation vector comprising the polynucleotide of claim 1 , wherein the heterologous promoter is functional in a plant cell.

13 . A cell transformed with the polynucleotide of claim 1 .

14 . The cell of claim 13 , wherein the cell is a prokaryotic cell.

15 . The cell of claim 13 , wherein the cell is a eukaryotic cell.

16 . The cell of claim 15 , wherein the cell is a plant cell.

17 . A plant transformed with the polynucleotide of claim 1 .

18 . A seed of the plant of claim 17 , wherein the seed comprises the polynucleotide.

19 . A commodity product produced from the plant of claim 17 , wherein the commodity product comprises a detectable amount of the polynucleotide.

20 . The plant of claim 17 , wherein the at least one polynucleotide is expressed in the plant as a double-stranded ribonucleic acid molecule.

21 . The cell of claim 16 , wherein the cell is a corn, soybean, or cotton cell.

22 . The plant of claim 17 , wherein the plant is corn, soybean, or cotton.

23 . The plant of claim 17 , wherein the at least one polynucleotide is expressed in the plant as a ribonucleic acid molecule, and the ribonucleic acid molecule inhibits the expression of an endogenous polynucleotide that is specifically complementary to the at least one polynucleotide when a coleopteran or hemipteran insect ingests a part of the plant.

24 . The polynucleotide of claim 1 , further comprising at least one additional polynucleotide that encodes a RNA molecule that inhibits the expression of an endogenous insect gene.

25 . A plant transformation vector comprising the polynucleotide of claim 24

Assignees

• Dow Agrosciences Llc
• Fraunhofer Ges Forschung

Inventors

• Narva Kenneth E
• Worden Sarah
• Frey Meghan
• Rangasamy Murugesan
• Gandra Premchand
• Veeramani Balaji
• Lo Wendy
• Fishilevich Elane
• Vilcinskas Andreas
• Knorr Eileen

Classifications

• C12N2310/531

  Stem-loop; Hairpin · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · 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