Parental rnai suppression of kruppel gene to control hemipteran pests

Patent metadata

Field	Value
Publication number	US-2016369296-A1
Application number	US-201514971550-A
Country	US
Kind code	A1
Filing date	Dec 16, 2015
Priority date	Dec 16, 2014
Publication date	Dec 22, 2016
Grant date	—

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.

First claim

What may be claimed is:

1 . An isolated nucleic acid comprising at least one polynucleotide 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 hemipteran organism comprising SEQ ID NO:1; the complement of a native coding sequence of a hemipteran organism comprising SEQ ID NO:1; a native non-coding sequence of a hemipteran organism that is transcribed into a native RNA molecule comprising SEQ ID NO:1; the complement of a native non-coding sequence of a hemipteran organism that is transcribed into a native RNA molecule comprising SEQ ID NO:1; a fragment of at least 15 contiguous nucleotides of a native coding sequence of a hemipteran organism comprising SEQ ID NO:1; the complement of a fragment of at least 15 contiguous nucleotides of a native coding sequence of a hemipteran organism comprising SEQ ID NO:1; a fragment of at least 15 contiguous nucleotides of a native non-coding sequence of a hemipteran organism that is transcribed into a native RNA molecule comprising SEQ ID NO:1; and the complement of a fragment of at least 15 contiguous nucleotides of a native non-coding sequence of a hemipteran organism that is transcribed into a native RNA molecule comprising SEQ ID NO:1, wherein the polynucleotide is operably linked to a heterologous promoter.

2 . The polynucleotide of claim 1 , wherein the polynucleotide is selected from the group consisting of SEQ ID NO:1; SEQ ID NO:3; and SEQ ID NO:4.

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

4 . The polynucleotide of claim 1 , wherein the organism is selected from the group consisting of Euschistus heros (Fabr.) (Neotropical Brown Stink Bug), Nezara viridula (L.) (Southern Green Stink Bug), Piezodorus guildinii (Westwood) (Red-banded Stink Bug), Halyomorpha halys (Stat) (Brown Marmorated Stink Bug), Chinavia hilare (Say) (Green Stink Bug), Euschistus serous (Say) (Brown Stink Bug), Dichelops melacanthus (Dallas), Dichelops furcatus (F.), Edessa meditabunda (F.), Thyanta perditor (F.) (Neotropical Red Shouldered Stink Bug), Chinavia marginatum (Palisot de Beauvois), Horcias nobilellus (Berg) (Cotton Bug), Taedia stigmosa (Berg), Dysdercus peruvianus (Guérin-Méneville), Neomegalotomus parvus (Westwood), Leptoglossus zonatus (Dallas), Niesthrea sidae (F.), Lygus hesperus (Knight) (Western Tarnished Plant Bug), and Lygus lineolaris (Palisot de Beauvois).

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

6 . A double-stranded ribonucleic acid molecule produced from the expression of the polynucleotide of claim 1 .

7 . The double-stranded ribonucleic acid molecule of claim 6 , wherein contacting the polynucleotide sequence with a hemipteran pest inhibits the expression of an endogenous nucleotide sequence specifically complementary to the polynucleotide.

8 . The double-stranded ribonucleic acid molecule of claim 7 , wherein contacting said ribonucleotide molecule with a hemipteran pest kills or inhibits the growth, reproduction, and/or feeding of the pest.

9 . The double stranded RNA of claim 6 , 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.

10 . The RNA of claim 5 , 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.

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

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

13 . The cell of claim 12 , wherein the cell is a prokaryotic cell.

14 . The cell of claim 12 , wherein the cell is a eukaryotic cell.

15 . The cell of claim 14 , wherein the cell is a plant cell.

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

17 . A seed of the plant of claim 16 , wherein the seed comprises the polynucleotide.

18 . A commodity product produced from the plant of claim 16 , wherein the commodity product comprises a detectable amount of the polynucleotide.

19 . The plant of claim 16 , wherein the at least one polynucleotide is expressed in the plant as a double-stranded ribonucleic acid molecule.

20 . The cell of claim 15 , wherein the cell is a Zea mays cell, a Glycine max cell, or a cell from a Gossypium sp.

21 . The plant of claim 16 , wherein the plant is maize, soybean, or cotton.

22 . The plant of claim 16 , 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 hemipteran pest ingests a part of the plant.

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

24 . The polynucleotide of claim 23 , wherein the additional polynucleotide encodes an iRNA molecule that results in a parental RNAi phenotype.

25 . The polynucleotide of claim 24 , wherein the additional polynucleotide encodes an iRNA molecule that inhibits the expression of a Brahma or hunchback gene.

26 . The polynucleotide of claim 23 , wherein the additional polynucleotide encodes an iRNA molecule that results in decreased growth and/or development and/or mortality in a hemipteran pest that contacts the iRNA molecule (lethal RNAi).

27 . A plant transformation vector comprising the polynucleotide of claim 23 , wherein the additional polynucleotide(s) are each operably linked to a heterologous promoter functional in a plant cell.

28 . 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:15-17; the complement of any of SEQ ID NOs:15-17; a fragment of at least 15 contiguous nucleotides of any of SEQ ID NOs:15-17; the complement of a fragment of at least 15 contiguous nucleotides of any of SEQ ID NOs:15-17; a transcript of SEQ ID NO:1; the complement of a transcript of SEQ ID NO:1; a fragment of at least 15 contiguous nucleotides of a transcript of SEQ ID NO:1; and the complement of a fragment of at least 15 contiguous nucleotides of a transcript of SEQ ID NO:1.

29 . The method according to claim 28 , wherein the agent is a double-stranded RNA molecule.

30 . A method for controlling a hemipteran pest population, the method comprising:
introducing i

Assignees

• Univ Nebraska
• Dow Agrosciences Llc

Inventors

• Siegfried Blair D
• Narva Kenneth E
• Arora Kanika
• Worden Sarah E
• Khajuria Chitvan
• Fishilevich Elane
• Storer Nicholas P
• Frey Meghan
• Hamm Ronda L
• Velez Ana

Classifications

• 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

• C12N15/8218 Primary

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

• C12N15/8286 Primary

  for insect resistance · CPC title

• A01N57/16

  containing heterocyclic radicals · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title