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RU2017113005A - Композиции, содержащие рекомбинантные клетки bacillus и инсектицид - Google Patents

Композиции, содержащие рекомбинантные клетки bacillus и инсектицид Download PDF

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RU2017113005A
RU2017113005A RU2017113005A RU2017113005A RU2017113005A RU 2017113005 A RU2017113005 A RU 2017113005A RU 2017113005 A RU2017113005 A RU 2017113005A RU 2017113005 A RU2017113005 A RU 2017113005A RU 2017113005 A RU2017113005 A RU 2017113005A
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composition
seq
targeting sequence
bacillus
protein
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RU2017113005A
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RU2736562C2 (ru
RU2017113005A3 (ru
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Деймиан КЁРТИС
Брайан ТОМПСОН
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Байер Кропсайенс Лп
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    • A01N63/22Bacillus
    • A01N63/23B. thuringiensis
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Claims (51)

1. Композиция, содержащая:
а) рекомбинантные продуцирующие экзоспорий клетки Bacillus, которые экспрессируют слитый белок, содержащий:
(I) по меньшей мере один белок или пептид, выбранный из группы, включающей стимулирующий рост растений белок или пептид и белок или пептид с инсектицидной активностью; и
(II) нацеливающую последовательность, белок экзоспория, или фрагмент белка экзоспория; и
б) по меньшей мере один инсектицид, выбранный из группы, включающей ацетамиприд, алдикарб, амитраз, бета-цифлутрин, карбарил, клотианидин, цифлутрин, циперметрин, дельтаметрин, эндосульфан, этион, этипрол, этопрофос, фенамифос, фенобукарб, фентион, фипронил, флубендиамид, флуопирам, флупирадифурон, форметанат, гептанофос, имидаклоприд, метамидофос, метиокарб, метомил, никлозамид, оксидеметон-метил, фосалон, силафлуофен, спиродиклофен, спиромезифен, спиротетрамат, тиаклоприд, тиодикарб, тралометрин, триазофос, трифлумурон, вамидотион, 1-{2-фтор-4-метил-5-[(R)-(2,2,2-трифторэтил)сульфинил]фенил}-3-(трифторметил)-1Н-1,2,4-триазол-5-амин, и 1-(3-хлорпиридин-2-ил)-N-[4-циано-2-метил-6-(метилкарбамоил)фенил]-3-{[5-(трифторметил)-2Н-тетразол-2-ил]метил}-1Н-пиразол-5-карбоксамид в синергетически эффективном количестве.
2. Композиция по п. 1, где по меньшей мере один белок или пептид представляет собой стимулирующий рост растений белок или пептид, выбранный из группы, включающей фермент, вовлеченный в продукцию или активацию соединения, стимулирующего рост растений, и фермент, который разлагает или модифицирует бактериальный, грибковый или растительный источник питания.
3. Композиция по п. 1, где продуцирующие экзоспорий клетки Bacillus представляют собой клетки представителя семейства Bacillus cereus.
4. Композиция в соответствии с пунктом 3, где представитель семейства Bacillus cereus выбирают из группы, включающей Bacillus anthracis, Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus pseudomycoides, Bacillus samanii, Bacillus gaemokensis, Bacillus weihenstephensis, Bacillus toyoiensis и их комбинации.
5. Композиция в соответствии с любым из пп. 1-4, где нацеливающая последовательность или белок экзоспория включает:
аминокислотную последовательность, имеющую по меньшей мере приблизительно 43% идентичность с аминокислотами 20-35 из SEQ ID NO: 1, где идентичность с аминокислотами 25-35 составляет по меньшей мере приблизительно 54%;
нацеливающую последовательность, содержащую аминокислоты 1-35 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 20-35 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 22-31 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 22-33 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 20-31 из SEQ ID NO: 1;
нацеливающая последовательность, содержащая SEQ ID NO: 1; или
белок экзоспория, содержащий аминокислотную последовательность, имеющую по меньшей мере 85% идентичность с SEQ ID NO: 2.
6. Композиция в соответствии с любым из пп. 2-5, где фермент, вовлеченный в продукцию или активацию соединения, стимулирующего рост растений, выбирают из группы, включающей ацетоин-редуктазу, индол-3-ацетамид-гидролазу, триптофан-монооксигеназу, ацетолактат-синтетазу, α-ацетолактат-декарбоксилазу, пируват-декарбоксилазу, диацетил-редуктазу, бутандиол-дегидрогеназу, аминотрансферазу, триптофан-декарбоксилазу, аминоксидазу, индол-3-пируват-декарбоксилазу, индол-3-ацетальдегид дегидрогеназу, оксидазу боковой цепи триптофана, нитрил-гидролазу, нитрилазу, пептидазу, протеазу, аденозинфосфат-изопентенил-трансферазу, фосфатазу, аденозин-киназу, аденин-фосфорибозилтрансферазу, CYP735A, 5'-рибонуклеотид-фосфогидролазу, аденозин-нуклеозидазу, зеатин цис-транс-изомеразу, зеатин О-гликозилтрансферазу, β-глюкозидазу, цис-гидроксилазу, CK цис-гидроксилазу, CK N-гликозилтрансферазу, 2,5-рибонуклеотид фосфогидролазу, аденозин-нуклеозидазу, пуриннуклеозид фосфорилазу, зеатин редуктазу, гидроксиламин редуктазу, 2-оксоглутарат диоксигеназу, гиббереллиновую 2В/3В гидролазу, гиббереллин 3-оксидазу, гиббереллин 20-оксидазу, хитозаназу, хитиназу, β-1,3- глюканазу, β-1,4-глюканазу, β-1,6-глюканазу, дезаминазу аминоциклопропан-1-карбоновой кислоты, и фермент, вовлеченный в продукцию nod-фактора.
7. Композиция по п. 6, где фермент, вовлеченный в продукцию или активацию соединения, стимулирующего рост растений, представляет собой хитозаназу.
8. Композиция по п. 7, где слитый белок включает SEQ ID NO: 109.
9. Композиция в соответствии с любым из пп. 2-5, где фермент, который разлагает или модифицирует бактериальный, грибковый или растительный источник питания, выбирают из группы, включающей целлюлазу, липазу, лигнин-оксидазу, протеазу, гликозид гидролазу, фосфатазу, нитрогеназу, нуклеазу, амидазу, нитрат-редуктазу, нитрит-редуктазу, амилазу, аммоний-оксидазу, лигниназу, глюкозидазу, фосфолипазу, фитазу, пектиназу, глюканазу, сульфатазу, уреазу, ксиланазу, и сидерофор.
10. Композиция по п. 9, где фермент представляет собой целлюлазу, выбранную из группы, включающей эндоцеллюлазу, экзоцеллюлазу и β-глюкозидазу.
11. Композиция по п. 10, где слитый белок включает эндоглюканазу Bacillus subtilis.
12. Композиция по п. 11, где слитый белок включает SEQ ID NO: 107.
13. Композиция по п. 12, где рекомбинантные клетки Bacillus имеют происхождение из Bacillus thuringiensis ВТ013А.
14. Композиция по п. 9, где фермент представляет собой фосфолипазу.
15. Композиция по п. 14, где слитый белок включает SEQ ID NO: 108.
16. Композиция в соответствии с любым из пп. 1-15, где слитый белок экспрессируется под контролем промотора спорообразования, нативного для нацеливающей последовательности, белка экзоспория, или фрагмента белка экзоспория слитого белка.
17. Композиция в соответствии с любым из пп. 1-16, где слитый белок экспрессируется под контролем высокоэкспрессируемого промотора спорообразования.
18. Композиция по п. 17, где высокоэкспрессируемый промотор спорообразования включает промоторную последовательность сигма-K полимеразы, специфической для спорообразования.
19. Композиция в соответствии с любым из пп. 16-18, где промотор спорообразования включает нуклеотидную последовательность, имеющую по меньшей мере 80% идентичность с нуклеотидной последовательностью с любой из SEQ ID NOS: 85-103.
20. Композиция в соответствии с любым из пп. 1-19, где по меньшей мере один инсектицид выбирают из группы, включающей клотианидин, циперметрин, этипрол, фипронил, флупирадифурон, и метиокарб.
21. Композиция по п. 20, где инсектицид представляет собой флупирадифурон.
22. Композиция в соответствии с любым из пп. 1-19, где инсектицид представляет собой клотианидин.
23. Семя, обработанное композицией в соответствии с любым из пп. 1-22.
24. Применение композиции в соответствии с любым из пп. 1-22 для усиления роста растения и/или способствования жизнеспособности растения.
25. Применение по п. 24 для обработки обычных или трансгенных растений или их семян.
26. Способ обработки растения, части растений, или локуса, окружающего растение, для усиления роста растения и/или способствования жизнеспособности растения, включающий стадию одновременного или последовательного использования:
а) рекомбинантных продуцирующих экзоспорий клеток Bacillus, которые экспрессируют слитый белок, содержащий:
(I) по меньшей мере один стимулирующий рост растений белок или пептид; и
(II) нацеливающую последовательность, белок экзоспория, или фрагмент белка экзоспория; и
б) по меньшей мере одного инсектицида, выбранного из группы, включающей ацетамиприд, алдикарб, амитраз, бета-цифлутрин, карбарил, клотианидин, цифлутрин, циперметрин, дельтаметрин, эндосульфан, этион, этипрол, этопрофос, фенамифос, фенобукарб, фентион, фипронил, флубендиамид, флуопирам, флупирадифурон, форметанат, гептанофос, имидаклоприд, метамидофос, метиокарб, метомил, никлозамид, оксидеметон-метил, фосалон, силафлуофен, спиродиклофен, спиромезифен, спиротетрамат, тиаклоприд, тиодикарб, тралометрин, триазофос, трифлумурон, вамидотион, 1-{2-фтор-4-метил-5-[(R)-(2,2,2-трифторэтил)сульфинил]фенил}-3-(трифторметил)-1Н-1,2,4-триазол-5-амин, и 1-(3-хлорпиридин-2-ил)-N-[4-циано-2-метил-6-(метилкарбамоил)фенил]-3-{[5-(трифторметил)-2Н-тетразол-2-ил]метил}-1Н-пиразол-5-карбоксамид в синергетически эффективном количестве.
27. Способ по п. 26, где нацеливающая последовательность или белок экзоспория включает:
аминокислотную последовательность, имеющую по меньшей мере приблизительно 43% идентичность с аминокислотами 20-35 из SEQ ID NO: 1, где идентичность с аминокислотами 25-35 составляет по меньшей мере приблизительно 54%;
нацеливающую последовательность, содержащую аминокислоты 1-35 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 20-35 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 22-31 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 22-33 из SEQ ID NO: 1;
нацеливающую последовательность, содержащую аминокислоты 20-31 из SEQ ID NO: 1;
нацеливающая последовательность, содержащая SEQ ID NO: 1; или
белок экзоспория, содержащий аминокислотную последовательность, имеющую по меньшей мере 85% идентичность с SEQ ID NO: 2.
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