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Biopesticide and insect preventing and controlling 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. Problems, achieve the effect of enhancing stability and efficacy, short cycle, enhancing stability and effect

Active Publication Date: 2010-12-22
GUANGZHOU RIBOBIO
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  • Summary
  • Abstract
  • Description
  • Claims
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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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  • Biopesticide and insect preventing and controlling method
  • Biopesticide and insect preventing and controlling method
  • Biopesticide and insect preventing and controlling method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] This example specifically provides the siRNA of the gene of the mitochondrial complex III Fe-S subunit of diamondback moth, which has a fatal effect on lepidopteran insects such as diamondback moth after natural feeding of the target gene siRNA.

[0030] This example is a method for controlling the lepidopteran insect diamondback moth (Plutella xylostella). By feeding siRNA containing silencing mitochondrial complex IIIFe-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.

[0031] 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 the membr...

Embodiment 2

[0064] This example particularly provides the siRNA of the mitochondrial complex IIIFe-S subunit gene of diamondback moth, by naturally feeding the target gene Si-UQCR_001 (SEQ ID NO 5 and 6) siRNA with the cationic polymer polyethyleneimine (PEI ), chitosan, polylysine and gelatin have caused fatal effects on lepidopteran insects such as diamondback moth.

[0065] Si-UQCR_001 (SEQ ID NO 1 and 2) siRNA and the above cationic polymer 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...

Embodiment 3

[0070] This example particularly provides the siRNA of the acetylcholinesterase gene of diamondback moth, which has a lethal effect on Lepidoptera insects such as diamondback moth after natural feeding of the siRNA of the target gene.

[0071] 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.

[0072] Acetylcholinesterase is a serine hydrolase whose main function is to interrupt 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 Pharmacol. Toxicol., 34:281-320)....

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Abstract

The invention provides an insect preventing and controlling method, which is to feed lepidopterous insets with a mixture of siRNA for the specific gene of the lepidopterous insets and a cationic polymer, wherein the siRNA is 17 to 50 nt double-chain RNA chemically synthesized in vitro; and the cationic polymer consists of polyethyleneimine (PEI), chitosan, polylysine and gelatin. The invention also relates to a biopesticide, which uses the siRNA for the specific gene of the lepidopterous insets as an active ingredient. In the invention, the gene expression of the lepidopterous insets is inhibited by feeding the lepidopterous insets with the siRNA which contains the specific gene of the lepidopterous insets, such as an interfering insect mitochondrion composite IIIFe-S subunit gene, a cholinesterase gene, a gamma-amino butyric acid receptor gene and the like, to interfere the normal growth and development of the insects, so the effective control of the insects is realized; and pot culture experiment result shows that the biopesticide is very effective.

Description

technical field [0001] 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 [0002] 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 (base) siRNA by Dicer family RNase-III, and the siRNA is further combined with Argonaute protein to form RISC (RNA-induced silencing complex), and finally RISC-mediated complementary combination of siRNA an...

Claims

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Application Information

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Patent Type & Authority Applications(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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