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Micromolecular RNA and application to pest control thereof

A pest control and small molecule technology, applied in the field of genetic engineering, can solve problems such as environmental pollution, food safety, and pest resistance

Active Publication Date: 2015-03-04
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the control of pests is still dominated by chemical pesticides, but a large number of long-term use not only caused the resistance of pests in various places, but also increased the use of pesticides, which in turn caused environmental pollution and affected food safety.

Method used

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  • Micromolecular RNA and application to pest control thereof
  • Micromolecular RNA and application to pest control thereof
  • Micromolecular RNA and application to pest control thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Five-instar first-day Spodoptera litura larvae with the same size and health status were selected, and the weight of each larva was weighed. They were randomly divided into miR-2771 mimics treatment group and NC mimics control group, with 40 larvae in each group, and repeated 3 times. Following the direction of blood circulation, miR-2771 mimics and NC mimics were injected from the flanks of Spodoptera litura larvae with a WPI microinjector, with an injection volume of 4 μg per larva, and then fed with mustard greens. On the 1st, 2nd, and 3rd day after injection, observe the morphological changes of the worms and take pictures to collect evidence, count the changes in the weight of the worms every day, and randomly select 3 worms to extract RNA and detect them by fluorescent quantitative PCR slgste1 mRNA expression.

[0022] figure 1 Show the body weight statistics of Spodoptera litura larvae after miRNAs injection, figure 2 It shows the appearance observation resu...

Embodiment 2

[0032] 1) Construct improved detection vectors IE1-pGL3-UTR and IE1-pGL3:

[0033] a) Insert the promoter IE1 upstream of the reporter luciferase gene in the pGL3 vector, and insert it downstream slgste1The sequence of the 3' non-coding region, the recombinant plasmid was transformed into DH5α bacteria, and the detection vector IE1-pGL3-UTR was constructed (such as Figure 4 shown in the middle left picture);

[0034] b) The promoter IE1 was inserted upstream of the reporter luciferase gene in the pGL3 vector, but not downstream slgste1 The 3' non-coding region sequence, the recombinant plasmid was transformed into DH5α bacteria, and the detection vector IE1-pGL3 was constructed;

[0035] 2) The day before transfection, transfer the Spli-221 cells in good growth condition into a twelve-well plate, add 1 mL of 10% FBS Grace's Insect medium and culture in a constant temperature incubator at 26°C overnight, when the cell density reaches 80% ~95%, carry out transfection opera...

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PUM

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Abstract

The invention discloses a small molecule RNA and its application in pest control. The present invention provides a small molecule RNA Sli-miR-2771. It is confirmed by in vivo and in vitro tests that Sli-miR-2771 can inhibit the expression of slgste1 by binding to the 3' non-coding region of detoxification enzyme glutathione transferase slgste1, so that Spodoptera litura reduces the ability to detoxify toxic plant secondary substances, and then affects the eating plants and growth status of Spodoptera litura, so as to achieve the purpose of pest control. The invention provides a new idea and a new approach for pest control, and will have broad application prospects in the field of pest control.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a small molecular RNA and its application in pest control. Background technique [0002] MicroRNA (miRNA) is a kind of non-coding small molecule RNA with a length of about 21-24 nucleotides, which widely exists in animals and plants. A series of experiments have shown that miRNA may be a class of molecules that are evolutionarily conserved and play important regulatory roles in organisms. [0003] miRNA is derived from the gene encoding miRNA in the nucleus and transcribed into pri-microRNA, which is then cleaved by Drosha enzyme into a hairpin-shaped pre-miRNA with a length of about 70 bp, and then transported from the nucleus to the cytoplasm by nucleoplasm / cytoplasm transport proteins Afterwards, it is cleaved by Dicer enzyme to grow into a 18-26bp miRNA double helix complex. One of them unwinds and incorporates into the RNA-induced gene silencing complex, formin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A01N57/16A01P7/04
Inventor 邹晓鹏郑思春冯启理邹海望徐智斌
Owner SOUTH CHINA NORMAL UNIVERSITY
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