COMPOSITIONS AND METHODS FOR CONTROLLING INSECT PESTS

• US 20170260522A1
• Filed: 07/28/2015
• Published: 09/14/2017
• Est. Priority Date: 07/29/2014
• Status: Active Grant

• Current Assignee: Monsanto Technology LLC (Bayer AG)
• Original Assignee: Monsanto Technology LLC (Bayer AG)
• Inventors: CRAWFORD, Michael John, EADS, Brian Donovan
• Granted Patent: US 10,378,012 B2
• Time in Patent Office: Days
• CPC Class Codes: A01N 57/16 containing heterocyclic radicals
• CPC Class Codes: A01N 63/60 Isolated nucleic acids
• CPC Class Codes: A01N 65/00 Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof containing compounds of determined constitution A01N27/00 - A01N59/00
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 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 Class Codes: C12N 15/8286 for insect resistance
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

Disclosed herein are polynucleotides, compositions, and methods for controlling insect pests, especially flea beetles, such as Phyllotreta spp. and Psylliodes spp., particularly in plants. More specifically, polynucleotides such as double-stranded RNA triggers and methods of use thereof for modifying the expression of genes in flea beetles.

Claims

1. A method of causing mortality or stunting in an insect, comprising providing in the diet of an insect at least one recombinant RNA comprising at least one silencing element essentially identical or essentially complementary to a fragment of a target gene sequence of said insect, wherein said target gene sequence is selected from the group consisting of SEQ ID NOs:

1-859, and wherein ingestion of said recombinant RNA by said insect results in mortality or stunting in said insect.

2. The method of claim 1, wherein said at least one silencing element comprises an RNA strand having a sequence of about 95% to about 100% identity or complementarity with a sequence selected from the group consisting of SEQ ID NOs:

860-1718 and 1722-1724.

3. The method of claim 1, wherein said insect is:

(a) a flea beetle;

(b) a species of a genus selected from the group consisting of the genera Altica, Anthobiodes, Aphthona, Aphthonaltica, Aphthonoides, Apteopeda, Argopistes, Argopus, Arrhenocoela, Batophila, Blepharida, Chaetocnema, Clitea, Crepidodera, Derocrepis, Dibolia, Disonycha, Epitrix, Hermipyxis, Hermaeophaga, Hespera, Hippuriphila, Horaia, Hyphasis, Lipromima, Liprus, Longitarsus, Luperomorpha, Lythraria, Manobia, Mantura, Meishania, Minota, Mniophila, Neicrepidodera, Nonarthra, Novofoudrasia, Ochrosis, Oedionychis, Oglobinia, Omeisphaera, Ophrida, Orestia, Paragopus, Pentamesa, Philopona, Phygasia, Phyllotreta, Podagrica, Podagricomela, Podontia, Pseudodera, Psylliodes, Sangariola, Sinaltica, Sphaeroderma, Systena, Trachyaphthona, Xuthea, and Zipangia;

(c) a species selected from the group consisting of Altica ambiens (alder flea beetle), Altica canadensis (prairie flea beetle), Altica chalybaea (grape flea beetle), Altica prasina (poplar flea beetle), Altica rosae (rose flea beetle), Altica sylvia (blueberry flea beetle), Altica ulmi (elm flea beetle), Chaetocnema pulicaria (corn flea beetle), Chaetocnema conofinis (sweet potato flea beetle), Epitrix cucumeris (potato flea beetle), Systena blanda (palestripped fleabeetle), and Systena frontalis (redheaded flea beetle);

(d) a species selected from the group consisting of Phyllotreta armoraciae (horseradish flea beetle), Phyllotreta cruciferae (canola flea beetle), Phyllotreta pusilla (western black flea beetle), Phyllotreta nemorum (striped turnip flea beetle), Phyllotreta atra (turnip flea beetle), Phyllotreta robusta (garden flea beetle), Phyllotreta striolata (striped flea beetle), Phyllotreta undulata, Psylliodes chrysocephala, and Psylliodes punctulata (hop flea beetle);

or

(e) an insect pest of a Brassica species.

4. The method of claim 1, wherein:

(a) said insect is a Phyllotreta species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-551;

(b) said insect is Phyllotreta atra (turnip flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-296;

(c) said insect is Phyllotreta cruciferae (canola flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

297-532;

(d) said insect is Phyllotreta striolata (striped flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

533-551;

(e) said insect is a Psylliodes species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859;

or

(f) said insect is Psylliodes chrysocephala and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859.

5. The method of claim 1, wherein said silencing element comprises:

(a) an RNA strand having at least one segment of 18 or more contiguous nucleotides with a sequence of 100% complementarity with a fragment of said target gene of said insect, wherein said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-859;

(b) an RNA strand having at least one segment of 18 or more contiguous nucleotides of an RNA sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724;

(c) an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724.

6. The method of claim 1, wherein:

(a) said insect is a Phyllotreta species and said silencing element comprises an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

860-1410;

(b) said insect is Phyllotreta atra (turnip flea beetle) and said silencing element comprises an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

860-1155;

said insect is Phyllotreta cruciferae (canola flea beetle) said silencing element comprises an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

1156-1391;

(d) said insect is Phyllotreta striolata (striped flea beetle) and said silencing element comprises an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

1392-1410;

(e) said insect is a Psylliodes species and said silencing element comprises an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

1411-1718;

or

(f) said insect is Psylliodes chrysocephala and said silencing element comprises an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

1411-1718.

7. The method of claim 1, wherein said recombinant RNA is provided in a microbial or plant cell that expresses said recombinant RNA, or in a microbial fermentation product, or is chemically synthesized.

8. The method of claim 1, wherein said silencing element comprises dsRNA.

10. The method of claim 1, wherein said at least one recombinant RNA is provided in a composition comprising said at least one recombinant RNA, wherein said composition is applied to a surface of said insect or to a surface of a seed or plant in need of protection from infestation by said insect.

11. The method of claim 10, wherein said composition:

(a) comprises a solid, liquid, powder, suspension, emulsion, spray, encapsulation, microbeads, carrier particulates, film, matrix, soil drench, or seed treatment;

(b) further comprises one or more components selected from the group consisting of a carrier agent, a surfactant, an organosilicone, a polynucleotide herbicidal molecule, a non-polynucleotide herbicidal molecule, a non-polynucleotide pesticide, a fungicide, a safener, a fertilizer, a micronutrient, an insect attractant, and an insect growth regulator;

(c) further comprises at least one pesticidal agent selected from the group consisting of a patatin, a plant lectin, a phytoecdysteroid, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, a bacterium that produces an insecticidal protein, an entomicidal bacterial species, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), Chromobacterium species, Chromobacterium subtsugae, Paenibacillus species, Paenibacillus lentimorbus, and Paenibacillus popilliae;

or

(d) is ingested by said insect.

9. The method of claim 9, wherein:

(a) said dsRNA is blunt-ended, or has an overhang at at least one terminus, or comprises at least one stem-loop;

(b) said dsRNA is chemically synthesized, produced by expression in a microorganism, produced by expression in a plant cell, or produced by microbial fermentation;

(c) said dsRNA is chemically modified;

or

(d) said dsRNA comprises at least one RNA strand having a sequence of about 95% to about 100% identity or complementarity with a sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724.

12. A method for controlling an insect infestation of a plant comprising contacting, with a dsRNA, an insect that infests a plant, wherein said dsRNA comprises at least one segment of 18 or more contiguous nucleotides with a sequence of about 95% to about 100% complementarity with a fragment of a target gene of said insect, and wherein said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs:

1-859.

13. The method of claim 12, wherein said dsRNA comprises:

(a) a sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724, or the complement thereof;

(b) at least one segment of 18 or more contiguous nucleotides with a sequence of 100% complementarity with a fragment of said target gene of said insect, wherein said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-859;

or

(c) at least one segment of 18 or more contiguous nucleotides of an RNA sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724.

14. The method of claim 12, wherein:

(a) said insect is a flea beetle;

(b) said insect is a species of a genus selected from the group consisting of the genera Altica, Anthobiodes, Aphthona, Aphthonaltica, Aphthonoides, Apteopeda, Argopistes, Argopus, Arrhenocoela, Batophila, Blepharida, Chaetocnema, Clitea, Crepidodera, Derocrepis, Dibolia, Disonycha, Epitrix, Hermipyxis, Hermaeophaga, Hespera, Hippuriphila, Horaia, Hyphasis, Lipromima, Liprus, Longitarsus, Luperomorpha, Lythraria, Manobia, Mantura, Meishania, Minota, Mniophila, Neicrepidodera, Nonarthra, Novofoudrasia, Ochrosis, Oedionychis, Oglobinia, Omeisphaera, Ophrida, Orestia, Paragopus, Pentamesa, Philopona, Phygasia, Phyllotreta, Podagrica, Podagricomela, Podontia, Pseudodera, Psylliodes, Sangariola, Sinaltica, Sphaeroderma, Systena, Trachyaphthona, Xuthea, and Zipangia;

(c) said insect is a species selected from the group consisting of Altica ambiens (alder flea beetle), Altica canadensis (prairie flea beetle), Altica chalybaea (grape flea beetle), Altica prasina (poplar flea beetle), Altica rosae (rose flea beetle), Altica sylvia (blueberry flea beetle), Altica ulmi (elm flea beetle), Chaetocnema pulicaria (corn flea beetle), Chaetocnema conofinis (sweet potato flea beetle), Epitrix cucumeris (potato flea beetle), Systena blanda (palestripped fleabeetle), and Systena frontalis (redheaded flea beetle);

(d) said insect is a species selected from the group consisting of Phyllotreta armoraciae (horseradish flea beetle), Phyllotreta cruciferae (canola flea beetle), Phyllotreta pusilla (western black flea beetle), Phyllotreta nemorum (striped turnip flea beetle), Phyllotreta atra (turnip flea beetle), Phyllotreta robusta (garden flea beetle), Phyllotreta striolata (striped flea beetle), Phyllotreta undulata, Psylliodes chrysocephala, and Psylliodes punctulata (hop flea beetle);

(e) said insect is an insect pest of a Brassica species;

(g) said insect is a Phyllotreta species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-551;

(h) said insect is Phyllotreta atra (turnip flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-296;

(i) said insect is Phyllotreta cruciferae (canola flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

297-532;

(j) said insect is Phyllotreta striolata (striped flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

533-551;

(k) said insect is a Psylliodes species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859;

(l) said insect is Psylliodes chrysocephala and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859;

(m) said insect is Phyllotreta atra (turnip flea beetle) and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

860-1155;

(n) said insect is Phyllotreta cruciferae (canola flea beetle) and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1156-1391;

(o) said insect is Phyllotreta striolata (striped flea beetle) and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1392-1410;

or

(p) said insect is Psylliodes chrysocephala and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1411-1718.

15. The method of claim 12, wherein said plant is:

(a) selected from an ornamental plant or a crop plant;

(b) a plant in the family Brassicaceae;

(c) a Brassica species selected from the group consisting of B. napus, B. juncea, B. carinata, B. rapa, B. oleracea, B. rupestris, B. septiceps, B. nigra, B. narinosa, B. perviridus, B. tournefortii, and B. fructiculosa;

(d) a Brassica plant selected from the group consisting canola, rapeseed, turnips, and field mustard or turnip rape;

(e) selected from the group consisting of Glycine max, Linum usitatissimum, Zea mays, Carthamus tinctorius, Helianthus annuus, Nicotiana tabacum, Arabidopsis thaliana, Betholettia excelsa, Ricinus communis, Cocos nucifera, Coriandrum sativum, Gossypium spp., Arachis hypogaea, Simmondsia chinensis, Solanum tuberosum, Elaeis guineensis, Olea europaea, Oryza sativa, Cucurbita maxim, Hordeum vulgare, and Triticum aestivum;

or

(f) a potato plant and said insect is Epitrix cucumeris (potato flea beetle).

16. The method of claim 12, wherein said contacting comprises:

(a) oral delivery to said insect, non-oral delivery to said insect, or a combination of oral and non-oral delivery to said insect;

(b) application of a composition comprising said dsRNA to a surface of said insect or to a surface of said plant infested by said insect;

(c) providing said dsRNA in a composition that further comprises one or more components selected from the group consisting of a carrier agent, a surfactant, an organosilicone, a polynucleotide herbicidal molecule, a non-polynucleotide herbicidal molecule, a non-polynucleotide pesticide, a fungicide, a safener, a fertilizer, a micronutrient, an insect attractant, and an insect growth regulator;

(d) providing said dsRNA in a composition that further comprises at least one pesticidal agent selected from the group consisting of a patatin, a plant lectin, a phytoecdysteroid, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, a bacterium that produces an insecticidal protein, an entomicidal bacterial species, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), Chromobacterium species, Chromobacterium subtsugae, Paenibacillus species, Paenibacillus lentimorbus, and Paenibacillus popilliae;

or

(e) providing said dsRNA in a composition that is ingested by said insect.

17. An insecticidal composition comprising an insecticidally effective amount of a recombinant RNA molecule, wherein said recombinant RNA molecule comprises at least one segment of 18 or more contiguous nucleotides with a sequence of about 95% to about 100% complementarity with a fragment of a target gene of an insect that infests a plant, and wherein said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs:

1-859.

18. The insecticidal composition of claim 17, wherein said recombinant RNA molecule:

(a) comprises at least one RNA strand having a sequence of about 95% to about 100% identity or complementarity with a sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724;

(b) comprises dsRNA;

(c) comprises dsRNA of at least 50 base pairs in length;

(d) comprises blunt-ended dsRNA;

(e) comprises dsRNA with an overhang at at least one terminus;

(f) comprises dsRNA comprising at least one stem-loop;

(g) comprises dsRNA that is chemically modified;

(h) is a dsRNA comprising an RNA strand having a sequence of about 95% to about 100% identity or complementarity with a sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724;

(i) is a dsRNA comprising an RNA strand having a sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724;

or

(j) is provided in a microbial or plant cell that expresses said recombinant RNA, or in a microbial fermentation product;

or is chemically synthesized.

19. The insecticidal composition of claim 17, wherein:

(a) said insect is a flea beetle;

(b) said insect is a species of a genus selected from the group consisting of the genera Altica, Anthobiodes, Aphthona, Aphthonaltica, Aphthonoides, Apteopeda, Argopistes, Argopus, Arrhenocoela, Batophila, Blepharida, Chaetocnema, Clitea, Crepidodera, Derocrepis, Dibolia, Disonycha, Epitrix, Hermipyxis, Hermaeophaga, Hespera, Hippuriphila, Horaia, Hyphasis, Lipromima, Liprus, Longitarsus, Luperomorpha, Lythraria, Manobia, Mantura, Meishania, Minota, Mniophila, Neicrepidodera, Nonarthra, Novofoudrasia, Ochrosis, Oedionychis, Oglobinia, Omeisphaera, Ophrida, Orestia, Paragopus, Pentamesa, Philopona, Phygasia, Phyllotreta, Podagrica, Podagricomela, Podontia, Pseudodera, Psylliodes, Sangariola, Sinaltica, Sphaeroderma, Systena, Trachyaphthona, Xuthea, and Zipangia;

(c) said insect is a species selected from the group consisting of Altica ambiens (alder flea beetle), Altica canadensis (prairie flea beetle), Altica chalybaea (grape flea beetle), Altica prasina (poplar flea beetle), Altica rosae (rose flea beetle), Altica sylvia (blueberry flea beetle), Altica ulmi (elm flea beetle), Chaetocnema pulicaria (corn flea beetle), Chaetocnema conofinis (sweet potato flea beetle), Epitrix cucumeris (potato flea beetle), Systena blanda (palestripped fleabeetle), and Systena frontalis (redheaded flea beetle);

(d) said insect is a species selected from the group consisting of Phyllotreta armoraciae (horseradish flea beetle), Phyllotreta cruciferae (canola flea beetle), Phyllotreta pusilla (western black flea beetle), Phyllotreta nemorum (striped turnip flea beetle), Phyllotreta atra (turnip flea beetle), Phyllotreta robusta (garden flea beetle), Phyllotreta striolata (striped flea beetle), Phyllotreta undulata, Psylliodes chrysocephala, and Psylliodes punctulata (hop flea beetle);

(e) said insect is an insect pest of a Brassica species;

(f) said insect is a Phyllotreta species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-551;

(g) said insect is Phyllotreta atra (turnip flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-296;

(h) said insect is Phyllotreta cruciferae (canola flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

297-532;

(i) said insect is Phyllotreta striolata (striped flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

533-551;

(j) said insect is a Psylliodes species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859;

(k) said insect is Psylliodes chrysocephala and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859;

(l) said insect is Phyllotreta atra (turnip flea beetle), said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

860-1155;

(m) said insect is Phyllotreta cruciferae (canola flea beetle), said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1156-1391;

(n) said insect is Phyllotreta striolata (striped flea beetle), said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1392-1410;

(o) said insect is Psylliodes chrysocephala, said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1411-1718;

(p) said plant is selected from an ornamental plant or a crop plant;

(q) said plant is a plant in the family Brassicaceae;

(r) said plant is a Brassica species selected from the group consisting of B. napus, B. juncea, B. carinata, B. rapa, B. oleracea, B. rupestris, B. septiceps, B. nigra, B. narinosa, B. perviridus, B. tournefortii, and B. fructiculosa;

(s) said plant is a Brassica plant selected from the group consisting canola, rapeseed, turnips, and field mustard or turnip rape;

(t) said plant is selected from the group consisting of Glycine max, Linum usitatissimum, Zea mays, Carthamus tinctorius, Helianthus annuus, Nicotiana tabacum, Arabidopsis thaliana, Betholettia excelsa, Ricinus communis, Cocos nucifera, Coriandrum sativum, Gossypium spp., Arachis hypogaea, Simmondsia chinensis, Solanum tuberosum, Elaeis guineensis, Olea europaea, Oryza sativa, Cucurbita maxim, Hordeum vulgare, and Triticum aestivum;

or

(u) said plant is a potato plant and said insect is Epitrix cucumeris (potato flea beetle).

20. The insecticidal composition of claim 17, wherein said insecticidal composition:

(a) further comprises one or more components selected from the group consisting of a carrier agent, a surfactant, an organosilicone, a polynucleotide herbicidal molecule, a non-polynucleotide herbicidal molecule, a non-polynucleotide pesticide, a fungicide, a safener, a fertilizer, a micronutrient, an insect attractant, and an insect growth regulator;

(b) further comprises at least one pesticidal agent selected from the group consisting of a patatin, a plant lectin, a phytoecdysteroid, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, a bacterium that produces an insecticidal protein, an entomicidal bacterial species, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), Chromobacterium species, Chromobacterium subtsugae, Paenibacillus species, Paenibacillus lentimorbus, and Paenibacillus popilliae;

or

(c) is in a form selected from the group consisting of a solid, liquid, powder, suspension, emulsion, spray, encapsulation, microbeads, carrier particulates, film, matrix, soil drench, insect diet or insect bait, and seed treatment.

21. A plant treated with the insecticidal composition of claim 17, or a plant grown from seed treated with the insecticidal composition of claim 17, wherein said plant exhibits improved resistance to said insect.

22. A method of providing a plant having improved resistance to an insect, comprising expressing in said plant a recombinant DNA construct, wherein said recombinant DNA construct comprises DNA encoding an RNA having a sequence essentially identical or essentially complementary to a fragment of at least one target gene of said insect, wherein said fragment comprises at least one segment of 18 or more contiguous nucleotides of said target gene, wherein said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs:

1-859, and wherein ingestion of said RNA by said insect results in mortality or stunting in said insect.

23. The method of claim 22, wherein said RNA:

(a) comprises a strand having a sequence of about 95% to about 100% identity or complementarity with a sequence selected from the group consisting of SEQ ID NOs;

860-1718 and 1722-1724;

or

(b) comprises dsRNA.

24. The method of claim 22, wherein:

(a) said insect is a flea beetle;

(b) said insect is a species of a genus selected from the group consisting of the genera Altica, Anthobiodes, Aphthona, Aphthonaltica, Aphthonoides, Apteopeda, Argopistes, Argopus, Arrhenocoela, Batophila, Blepharida, Chaetocnema, Clitea, Crepidodera, Derocrepis, Dibolia, Disonycha, Epitrix, Hermipyxis, Hermaeophaga, Hespera, Hippuriphila, Horaia, Hyphasis, Lipromima, Liprus, Longitarsus, Luperomorpha, Lythraria, Manobia, Mantura, Meishania, Minota, Mniophila, Neicrepidodera, Nonarthra, Novofoudrasia, Ochrosis, Oedionychis, Oglobinia, Omeisphaera, Ophrida, Orestia, Paragopus, Pentamesa, Philopona, Phygasia, Phyllotreta, Podagrica, Podagricomela, Podontia, Pseudodera, Psylliodes, Sangariola, Sinaltica, Sphaeroderma, Systena, Trachyaphthona, Xuthea, and Zipangia;

(c) said insect is a species selected from the group consisting of Altica ambiens (alder flea beetle), Altica canadensis (prairie flea beetle), Altica chalybaea (grape flea beetle), Altica prasina (poplar flea beetle), Altica rosae (rose flea beetle), Altica sylvia (blueberry flea beetle), Altica ulmi (elm flea beetle), Chaetocnema pulicaria (corn flea beetle), Chaetocnema conofinis (sweet potato flea beetle), Epitrix cucumeris (potato flea beetle), Systena blanda (palestripped fleabeetle), and Systena frontalis (redheaded flea beetle);

(d) said insect is a species selected from the group consisting of Phyllotreta armoraciae (horseradish flea beetle), Phyllotreta cruciferae (canola flea beetle), Phyllotreta pusilla (western black flea beetle), Phyllotreta nemorum (striped turnip flea beetle), Phyllotreta atra (turnip flea beetle), Phyllotreta robusta (garden flea beetle), Phyllotreta striolata (striped flea beetle), Phyllotreta undulata, Psylliodes chrysocephala, and Psylliodes punctulata (hop flea beetle);

(e) said insect is an insect pest of a Brassica species;

(f) said insect is a Phyllotreta species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-551;

(g) said insect is Phyllotreta atra (turnip flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

1-296;

(h) said insect is Phyllotreta cruciferae (canola flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

297-532;

(i) said insect is Phyllotreta striolata (striped flea beetle) and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

533-551;

(j) said insect is a Psylliodes species and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859;

(k) said insect is Psylliodes chrysocephala and said target gene has a DNA sequence selected from the group consisting of SEQ ID NOs;

552-859;

(l) said insect is Phyllotreta atra (turnip flea beetle), said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

860-1155;

(m) said insect is Phyllotreta cruciferae (canola flea beetle), said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1156-1391;

(n) said insect is Phyllotreta striolata (striped flea beetle), said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1392-1410;

(o) said insect is Psylliodes chrysocephala, said recombinant RNA molecule comprises dsRNA, and said dsRNA comprises at least one RNA sequence selected from the group consisting of SEQ ID NOs;

1411-1718;

(p) said plant is selected from an ornamental plant or a crop plant;

(q) said plant is a plant in the family Brassicaceae;

(r) said plant is a Brassica species selected from the group consisting of B. napus, B. juncea, B. carinata, B. rapa, B. oleracea, B. rupestris, B. septiceps, B. nigra, B. narinosa, B. perviridus, B. tournefortii, and B. fructiculosa;

(s) said plant is a Brassica plant selected from the group consisting canola, rapeseed, turnips, and field mustard or turnip rape;

(t) said plant is selected from the group consisting of Glycine max, Linum usitatissimum, Zea mays, Carthamus tinctorius, Helianthus annuus, Nicotiana tabacum, Arabidopsis thaliana, Betholettia excelsa, Ricinus communis, Cocos nucifera, Coriandrum sativum, Gossypium spp., Arachis hypogaea, Simmondsia chinensis, Solanum tuberosum, Elaeis guineensis, Olea europaea, Oryza sativa, Cucurbita maxim, Hordeum vulgare, and Triticum aestivum;

(u) said plant is a potato plant and said insect is Epitrix cucumeris (potato flea beetle);

(v) said method further comprises expression in said plant of at least one pesticidal agent selected from the group consisting of a patatin, a plant lectin, a phytoecdysteroid, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, a Bacillus sphaericus insecticidal protein, a bacterium that produces an insecticidal protein, an entomicidal bacterial species, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), Chromobacterium species, Chromobacterium subtsugae, Paenibacillus species, Paenibacillus lentimorbus, and Paenibacillus popilliae;

or

(w) said method further comprises expression in said plant of at least one protein conferring tolerance to an herbicide.

25. The plant having improved resistance to an insect, provided by the method of claim 22, or fruit, seed, or propagatable parts of said plant.

26. A recombinant DNA construct comprising a heterologous promoter operably linked to DNA encoding an RNA transcript comprising a sequence of about 95% to about 100% identity or complementarity with a sequence selected from the group consisting of SEQ ID NOs:

860-1718 and 1722-1724.

27. The recombinant DNA construct of claim 26, wherein said heterologous promoter is functional for expression of said RNA transcript in a bacterium or in a plant cell.

28. A recombinant vector, plant chromosome or plastid, or transgenic plant cell comprising the recombinant DNA construct of claim 26.