RU2021123470A - Композиции и способы борьбы с насекомыми-вредителями - Google Patents

Композиции и способы борьбы с насекомыми-вредителями Download PDF

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RU2021123470A
RU2021123470A RU2021123470A RU2021123470A RU2021123470A RU 2021123470 A RU2021123470 A RU 2021123470A RU 2021123470 A RU2021123470 A RU 2021123470A RU 2021123470 A RU2021123470 A RU 2021123470A RU 2021123470 A RU2021123470 A RU 2021123470A
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insect
plant
dsrna
phyllotreta spp
phyllotreta
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Майкл Джон КРОУФОРД
Брайан Донован ИДС
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Монсанто Текнолоджи Ллс
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    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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    • A01N57/10Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds
    • A01N57/16Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds containing heterocyclic radicals
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    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • C12N15/8286Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for insect resistance
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Claims (67)

1. Способ умерщвления насекомого Phyllotreta spp., включающий предоставление в рацион насекомого Phyllotreta spp. по меньшей мере одной рекомбинантной двухцепочечной РНК (дцРНК), содержащей по меньшей мере один подавляющий экспрессию элемент, содержащий цепь РНК, идентичную или комплементарную 21 или более смежным нуклеотидам в SEQ ID NO: 335 и ее комплемент, и, где поглощение указанной рекомбинантной дцРНК указанным насекомым Phyllotreta spp. приводит к гибели указанного насекомого Phyllotreta spp..
2. Способ задержки развития насекомого Phyllotreta spp., включающий предоставление в рацион насекомого Phyllotreta spp. по меньшей мере одной рекомбинантной дцРНК, содержащей по меньшей мере один подавляющий экспрессию элемент, содержащий цепь РНК, идентичную или комплементарную 21 или более смежным нуклеотидам в SEQ ID NO: 335 и ее комплемент, и, где поглощение указанной рекомбинантной дцРНК указанным насекомым Phyllotreta spp. приводит к задержке развития указанного насекомого Phyllotreta spp..
3. Способ по п. 1 или 2, отличающийся тем, что указанный по меньшей мере один подавляющий экспрессию элемент содержит цепь РНК, имеющую последовательность, идентичную или комплементарную 21 или более смежным нуклеотидам в SEQ ID NO: 1193.
4. Способ по п. 1 или 2, отличающийся тем, что указанное насекомое Phyllotreta spp. представляет собой Phyllotreta cruciferae (блошки канолы).
5. Способ по п. 1 или 2, отличающийся тем, что указанный подавляющий экспрессию элемент содержит цепь РНК, обладающую последовательностью SEQ ID NO: 1193.
6. Способ по п. 1 или 2, отличающийся тем, что указанное насекомое Phyllotreta spp. представляет собой Phyllotreta cruciferae (блошку канолы), и указанный подавляющий экспрессию элемент содержит цепь РНК, имеющую последовательность SEQ ID NO: 1193.
7. Способ по п. 1 или 2, отличающийся тем, что указанная рекомбинантная дцРНК представлена в микробной или растительной клетке, которая экспрессирует указанную рекомбинантную РНК, или в микробном продукте ферментации, или является химически синтезированной.
8. Способ по п. 1 или 2, отличающийся тем, что:
(a) указанная дцРНК является тупоконечной или имеет "липкие" концы по меньшей мере на одном конце, или содержит по меньшей мере один стебель-петлю;
(b) указанная дцРНК является химически синтезированной, полученной путем экспрессии в микроорганизме, полученной путем экспрессии в растительной клетке, или полученной путем микробной ферментации; или
(c) указанная дцРНК является химически синтезированной.
9. Способ по п. 1 или 2, отличающийся тем, что указанная по меньшей мере одна рекомбинантная дцРНК представлена в композиции, содержащей указанную по меньшей мере одну рекомбинантную дцРНК, причем указанную композицию наносят на поверхность указанного насекомого Phyllotreta spp. или на поверхность семян или растений, нуждающихся в защите от заражения указанным насекомым Phyllotreta spp..
10. Способ по п. 9, отличающийся тем, что указанная композиция:
(a) содержит твердое вещество, жидкость, порошок, суспензию, эмульсию, спрей, инкапсуляцию, микрогранулы, частицы носителей, пленки, матрицы, пропитку почвы или обработку семян;
(b) дополнительно содержит один или более компонентов, выбранных из группы, состоящей из агента-переносчика, поверхностно-активного вещества, кремнийорганики, полинуклеотидной гербицидной молекулы, неполинуклеотидной гербицидной молекулы, неполинуклеотидного пестицида, фунгицида, антидота, удобрения, микроэлемента, аттрактанта насекомых и регулятора роста насекомых;
(c) дополнительно содержит по меньшей мере один пестицидный агент, выбранный из группы, состоящей из пататина, растительного лектина, фитоэкдистероида, инсектицидного белка Bacillus thuringiensis, инсектицидного белка Xenorhabdus, инсектицидного белка Photorhabdus, инсектицидного белка Bacillus laterosporous, инсектицидного белка Bacillus sphaericus, бактерии, которая производит инсектицидный белок, энтомицидных видов бактерий, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), видов Chromobacterium, Chromobacterium subtsugae, видов Paenibacillus, Paenibacillus lentimorbus и Paenibacillus popilliae; или
(d) поглощается указанным насекомым Phyllotreta spp..
11. Способ борьбы с насекомым Phyllotreta spp., заражающим растение, включающий контакт дцРНК с насекомым Phyllotreta spp., которое заражает растение, причем указанная дцРНК содержит цепь РНК, идентичную или комплементарную 21 или более смежным нуклеотидам последовательности гена-мишени, которая имеет SEQ ID NO: 335 и ее комплемент.
12. Способ по п. 11, отличающийся тем, что указанная дцРНК содержит:
(a) последовательность SEQ ID NO: 1193, или ее комплемент; или
(b) цепь РНК, идентичную или комплементарную 21 или более смежным нуклеотидам в SEQ ID NO:1193.
13. Способ по п. 11, отличающийся тем, что указанное насекомое Phyllotreta spp. представляет собой Phyllotreta cruciferae (блошку канолы.
14. Способ по п. 11, отличающийся тем, что указанное растение представляет собой:
(a) выбранное из декоративного растения или сельскохозяйственного растения;
(b) растение из семейства Brassicaceae;
(c) вид рода Brassica, выбранный из группы, состоящей из B. napus, B. juncea, B. carinata, B. rapa, B. oleracea, B. rupestris, B. septiceps, B. nigra, B. narinosa, B. perviridus, B. tournefortii и B. fructiculosa;
(d) растение Brassica, выбранное из группы, включающей канолу, рапс, репы и горчицу полевую или репу масличную; или
(e) выбранное из группы, состоящей из 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 maxima, Hordeum vulgare и Triticum aestivum.
15. Способ по п. 11, отличающийся тем, что стадия, в которой осуществляют контакт, включает:
(a) пероральное введение указанному насекомому Phyllotreta spp., непероральное введение указанному насекомому Phyllotreta spp., или комбинацию перорального и неперорального введения указанному насекомому Phyllotreta spp.;
(b) нанесение композиции, содержащей указанную дцРНК, на поверхность указанного насекомого Phyllotreta spp. или на поверхность указанного растения, заражаемого указанным насекомым Phyllotreta spp.;
(c) предоставление указанной дцРНК в композиции, которая дополнительно содержит один или более компонентов, выбранных из группы, состоящей из агента-переносчика, поверхностно-активного вещества, кремнийорганики, полинуклеотидной гербицидной молекулы, неполинуклеотидной гербицидной молекулы, неполинуклеотидного пестицида, фунгицида, антидота, удобрения, микроэлемента, аттрактанта насекомых и регулятора роста насекомых;
(d) предоставление указанной дцРНК в композиции, которая дополнительно содержит по меньшей мере один пестицидный агент, выбранный из группы, состоящей из пататина, растительного лектина, фитоэкдистероида, инсектицидного белка Bacillus thuringiensis, инсектицидного белка Xenorhabdus, инсектицидного белка Photorhabdus, инсектицидного белка Bacillus laterosporous, инсектицидного белка Bacillus sphaericus, бактерии, которая производит инсектицидный белок, энтомицидных видов бактерий, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), видов Chromobacterium, Chromobacterium subtsugae, видов Paenibacillus, Paenibacillus lentimorbus и Paenibacillus popilliae; или
(e) предоставление указанной дцРНК в композиции, которая поглощается указанным насекомым Phyllotreta spp..
16. Инсектицидная композиция, обладающая инсектицидной активностью против Phyllotreta spp., содержащая инсектицидно-эффективное количество молекулы рекомбинантной дцРНК, причем указанная молекула рекомбинантной дцРНК содержит цепь РНК, идентичную или комплементарную 21 или более смежным нуклеотидам гена-мишени насекомого Phyllotreta spp., которое заражает растение, причем указанный ген-мишень имеет последовательность ДНК SEQ ID NO: 335.
17. Инсектицидная композиция по п. 16, отличающаяся тем, что указанная молекула рекомбинантной дцРНК:
(a) содержит по меньшей мере одну цепь РНК, идентичную или комплементарную 21 или более смежным нуклеотидам в SEQ ID NO: 1193;
(b) содержит дцРНК из по меньшей мере 50 пар оснований в длину;
(c) содержит тупоконечную дцРНК;
(d) содержит дцРНК с "липким" концом на по меньшей мере одном конце;
(e) содержит дцРНК, содержащую по меньшей мере один стебель-петлю;
(f) содержит дцРНК, которая является химически синтезированной;
(g) представляет собой дцРНК, содержащую цепь РНК, идентичную или комплементарную 21 или более смежным нуклеотидам в SEQ ID NO: 1193;
(h) представляет собой дцРНК, содержащую цепь РНК, имеющую последовательность SEQ ID NO: 1193; или
(i) представлена в микробной или растительной клетке, которая экспрессирует указанную рекомбинантную дцРНК, или в микробном продукте ферментации, или является химически синтезированной.
18. Инсектицидная композиция по п. 16, отличающаяся тем, что указанное насекомое Phyllotreta spp. представляет собой Phyllotreta cruciferae (блошку канолы).
19. Инсектицидная композиция по п. 16, отличающаяся тем, что указанная инсектицидная композиция:
(a) дополнительно содержит один или более компонентов, выбранных из группы, состоящей из агента-переносчика, поверхностно-активного вещества, кремнийорганики, полинуклеотидной гербицидной молекулы, неполинуклеотидной гербицидной молекулы, неполинуклеотидного пестицида, фунгицида, антидота, удобрения, микроэлемента, аттрактанта насекомых и регулятора роста насекомых;
(b) дополнительно содержит по меньшей мере один пестицидный агент, выбранный из группы, состоящей из пататина, растительного лектина, фитоэкдистероида, инсектицидного белка Bacillus thuringiensis, инсектицидного белка Xenorhabdus, инсектицидного белка Photorhabdus, инсектицидного белка Bacillus laterosporous, инсектицидного белка Bacillus sphaericus, бактерии, которая производит инсектицидный белок, энтомицидных видов бактерий, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), видов Chromobacterium, Chromobacterium subtsugae, видов Paenibacillus, Paenibacillus lentimorbus и Paenibacillus popilliae; или
(c) находится в форме, выбранной из группы, состоящей из твердого вещества, жидкости, порошка, суспензии, эмульсии, спрея, инкапсуляции, микрогранул, частиц носителей, пленки, матрицы, пропитку почвы, рациона насекомых или приманки насекомых и обработки семян.
20. Растение, обработанное инсектицидной композицией по п. 16, где указанная молекула рекомбинантной дцРНК проникает в клетки указанного растения, и, где указанное растение проявляет повышенную устойчивость к указанному насекомому Phyllotreta spp. по сравнению с растением, не обработанным инсектицидной композицией по п. 16.
21. Способ, обеспечивающий растение, имеющее повышенную стойкость к насекомому Phyllotreta spp., включающий (a) подготовку рекомбинантного ДНК конструкта, содержащего ДНК, кодирующую дцРНК, имеющую последовательность, идентичную или комплементарную 21 или более смежным нуклеотидам SEQ ID NO: 335, и (b) экспрессию указанного рекомбинантного ДНК конструкта в указанном растении для продуцирования указанной дцРНК, где поглощение указанной дцРНК указанным насекомым Phyllotreta spp. приводит к гибели указанного насекомого Phyllotreta spp., таким образом, обеспечивая указанное растение повышенной стойкостью к указанному насекомому Phyllotreta spp. по сравнению с растением, не экспрессирующим указанный рекомбинантный ДНК конструкт.
22. Способ, обеспечивающий растение, имеющее повышенную стойкость к насекомому Phyllotreta spp., включающий (a) подготовку рекомбинантного ДНК конструкта, содержащего ДНК, кодирующую дцРНК, имеющую последовательность идентичную или комплементарную 21 или более смежным нуклеотидам SEQ ID NO: 335, и (b) экспрессию указанного рекомбинантного ДНК конструкта в указанном растении для продуцирования указанной дцРНК, где поглощение указанной дцРНК указанным насекомым Phyllotreta spp. приводит к задержке развития указанного насекомого Phyllotreta spp., таким образом, обеспечивая указанное растение повышенной стойкостью к указанному насекомому Phyllotreta spp. по сравнению с растением, не экспрессирующим указанный рекомбинантный ДНК конструкт.
23. Способ по п. 21 или 22, отличающийся тем, что указанная дцРНК содержит цепь, имеющую последовательность идентичную или комплементарную 21 или более смежным нуклеотидам в SEQ ID NO: 1193.
24. Способ по п. 21 или 22, отличающийся тем, что:
(a) указанное насекомое Phyllotreta spp. представляет собой Phyllotreta cruciferae (блошку канолы)
(b) указанное растение является выбранным из декоративного растения или сельскохозяйственного растения;
(c) указанное растение представляет собой растение из семейства Brassicaceae;
(d) указанное растение представляет собой вид Brassica, выбранный из группы, состоящей из B. napus, B. juncea, B. carinata, B. rapa, B. oleracea, B. rupestris, B. septiceps, B. nigra, B. narinosa, B. perviridus, B. tournefortii, и B. fructiculosa;
(e) указанное растение представляет собой растение Brassica, выбранное из группы, содержащей канолу, рапс, репу и горчицу полевую или репу масличную;
(f) указанное растение является выбранным из группы, состоящей из 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 maxima, Hordeum vulgare и Triticum aestivum;
(g) указанный способ дополнительно включает экспрессию в указанном растении по меньшей мере одного пестицидного агента, выбранного из группы, состоящей из пататина, растительного лектина, фитоэкдистероида, инсектицидного белка Bacillus thuringiensis , инсектицидного белка Xenorhabdus, инсектицидного белка Photorhabdus, инсектицидного белка Bacillus laterosporous, инсектицидного белка Bacillus sphaericus, бактерии, которая производит инсектицидный белок, энтомицидных видов бактерий, Lysinibacillus sphaericus (Bacillus sphaericus), Brevibacillus laterosporus (Bacillus laterosporus), видов Chromobacterium, Chromobacterium subtsugae, видов Paenibacillus, Paenibacillus lentimorbus и Paenibacillus popilliae; или
(h) указанный способ дополнительно включает экспрессию в указанном растении по меньшей мере одного белка, придающего устойчивость к гербициду.
25. Растение, имеющее повышенную устойчивость к насекомому Phyllotreta spp., обеспеченное способом по п. 21 или 22, где указанное растение экспрессирует рекомбинантный ДНК конструкт, содержащий ДНК, кодирующую дцРНК, имеющую последовательность, идентичную или комплементарную 21 или более смежным нуклеотидам SEQ ID NO: 335, и, где указанное растение обладает повышенной устойчивостью к указанному насекомому Phyllotreta spp. по сравнению с растением, не экспрессирующим указанный рекомбинантный ДНК конструкт.
26. Рекомбинантный ДНК конструкт для борьбы с насекомым Phyllotreta spp., заражающим растение, содержащий гетерологичный промотор функционально связанный с ДНК, кодирующей РНК транскрипт, содержащий последовательность, идентичную или комплементарную 21 смежным нуклеотидам в SEQ ID NO: 1193.
27. Рекомбинантный ДНК конструкт по п. 26, отличающийся тем, что указанный гетерологичный промотор является функционирующим для экспрессирования указанного РНК транскрипта в бактерии или в растительной клетке.
28. Трансгенная растительная клетка для обеспечения повышенной устойчивости к насекомому Phyllotreta spp., где указанная трансгенная клетка содержит рекомбинантный ДНК конструкт по п. 26.
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