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A kind of biological pesticide and insect control method

A biopesticide and lepidopteran insect technology, applied in botany equipment and methods, insecticides, biocides, etc., can solve the problem of slow effect of engineering strains expressing dsRNA, food safety problems of transgenic plants, high cost of dsRNA synthesis, etc. problem, to achieve the effect of short cycle, enhanced stability and effect, and simple method

Active Publication Date: 2014-10-22
GUANGZHOU RIBOBIO
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AI Technical Summary

Problems solved by technology

At present, the major commonality and key technologies that restrict the use of RNAi technology to produce new biopesticides are: 1. The existing technical systems and strategies that use transgenic technology to transfer crops to control pests have food safety problems for transgenic plants; 2. Engineering strains expressing dsRNA The effect is slow, the control effect is low, and the environmental adaptability is difficult to evaluate; 3. The cost of in vitro transcription and synthesis of dsRNA is high, and the stability is not good, etc.
These problems greatly limit the wide application of this technology in the field of pest control.

Method used

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  • A kind of biological pesticide and insect control method
  • A kind of biological pesticide and insect control method
  • A kind of biological pesticide and insect control method

Examples

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Effect test

Embodiment 1

[0034] This example specifically provides the siRNA of the mitochondrial complex III Fe-S subunit gene of Plutella xylostella, which has a fatal effect on Lepidoptera insects such as Plutella xylostella after naturally feeding the siRNA of the target gene.

[0035] This example is a method for controlling the lepidopteran insect diamondback moth (Plutella xylostella). By feeding siRNA containing the silencing mitochondrial complex III Fe-S subunit gene (GeneBank accession number EU815629), the insect mitochondrial electron transfer is blocked, and the formation of insect ATP is inhibited, thereby achieving effective control of insects.

[0036] Mitochondrial complex III, also known as ubiquinol-cytochrome c reductase, consists of 9-11 subunits, including two cytochrome b (b562 and b566), one cytochrome C1 and one Fe- S protein. Among them, the Fe-S protein subunit is responsible for transferring an electron from reduced coenzyme Q to cytochrome C1, and releasing a proton to t...

Embodiment 2

[0068] This example specifically provides siRNA of the mitochondrial complex III Fe-S subunit gene of Plutella xylostella, by naturally feeding the target gene Si-UQCR_001 (SEQ ID NO 5 and 6) siRNA with cationic polymer polyethyleneimine (PEI ), chitosan, polylysine and gelatin have caused fatal effects on lepidopteran insects such as diamondback moth.

[0069]Si-UQCR_001 (SEQ ID NO 1 and 2) siRNA and the above cationic polymers were prepared into biopesticides, and evenly coated on cabbage leaves and fed to the second instar larvae of Plutella xylostella. In groups 1-4, N in biopesticides :P is 1:1, and in groups 5-6, N:P in biopesticides is 20:1. The leaf concentration of siRNA was 3.0 μg / cm 2 . 30 worms were treated with each pair of siRNA, and three replicates were set up, and DEPC water was used as a negative control group. After that, the number of dead insects was investigated respectively at 12h, 24h, 36h, 48h, 60h, and 72h, and the mortality rate was calculated. Th...

Embodiment 3

[0075] This example specifically provides siRNA of the acetylcholinesterase gene of diamondback moth, which has caused fatal effects on lepidopteran insects such as diamondback moth after natural feeding of the siRNA of the target gene.

[0076] This example is a method for controlling the lepidopteran insect diamondback moth (Plutella xylostella). By feeding siRNA containing silencing acetylcholinesterase genes 1 and 2 (GeneBank accession numbers: AY970293 and AY061975), the normal nerve impulse transmission of insects is affected, and the insects continue to be in a state of excitement and die, thereby realizing the effective treatment of insects. governance.

[0077] Acetylcholinesterase is a serine hydrolase whose main function is to stop the transmission of nerve impulses by rapidly hydrolyzing the neurotransmitter acetylcholine (ACh) at cholinergic synapses (Taylor T, Radie Z, 1994. Thecholinesterases: from genes to proteins.Annu Rev Pharmaco1.Toxico1., 34:281-320). Th...

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Abstract

The invention provides an insect control method, wherein siRNA specific to lepidopteron specific genes is matched with cation polymer to feed lepidopteron, siRNA is double-chain RNA of 17-50nt formed by in-nitro chemical synthesis, and cation polymer is polyethyleneimine (PEI), chitosan, polylysine and gelatin. The invention also relates to a biopesticide which takes siRNA specific to lepidopteron specific genes as the effective component. According to the invention, by restraining genetic expression of siRNA containing specific insect genes such as insect mitochondria interfering compound II Fe-S subunit gene, acetylcholine esterase gene, gamma-aminobutyric acid receptor gene and the like on lepidopteron, the normal growth and development of insect are interfered, so as to realize effective control on insects; and further shown by result of a pot culture test, the biopesticide is an efficient biopesticide.

Description

[0001] Original application date: 2010-3-5 [0002] Original application number: 201010122016.1 [0003] Title of original application for invention: A biological pesticide and insect control method technical field [0004] The invention relates to the field of chemically synthesized and modified small molecule interference RNA, in particular to a biological pesticide and insect control method. Background technique [0005] RNA interference (RNA interference, RNAi) refers to the phenomenon that exogenous or endogenous double-stranded RNA (dsRNA) specifically induces gene expression silencing. It is a gene regulation mechanism that is highly conserved in evolution and ubiquitous in the biological world. . First discovered in 1998 in the nematode Caenorhabditis elegans, the phenomenon has since been found in fungi, plants, insects and animals. The mechanism of RNAi is: the long-chain dsRNA introduced in vitro or generated by endogenous transcription is cut into 21-25nt (bas...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N57/16A01N61/00A01N43/44A01N43/16A01N37/46A01N43/90A01P7/04C12N15/113
Inventor 张必良龚亮胡美英
Owner GUANGZHOU RIBOBIO
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