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Artificially designed and synthesized miRNA and pest control action thereof on aphids

A kind of aphid and molecular technology, applied in DNA/RNA fragments, recombinant DNA technology, applications, etc., can solve the problem of lack of anti-aphid genes, and achieve the effect of high research value and broad application prospects.

Active Publication Date: 2017-07-14
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, due to the lack of effective aphid resistance genes in the existing germplasm resources, it is urgent for researchers to develop new target genes for the control of wheat pests

Method used

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  • Artificially designed and synthesized miRNA and pest control action thereof on aphids
  • Artificially designed and synthesized miRNA and pest control action thereof on aphids
  • Artificially designed and synthesized miRNA and pest control action thereof on aphids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Design and synthesis of miRNA avi-miR-316 agomir and miRNA avi-miR-316 antagomir

[0026] The corresponding miRNA avi-miR-316agomir (miRNA agonist) and miRNA avi-miR-316 antagomir (miRNA antagonist) were designed based on the mature sequence of aphid miRNA avi-miR-316. The miRNAavi-miR-316 agomir has a double-stranded structure, and the nucleotide sequences of the two strands are shown in SEQ ID NO: 1 in the sequence listing and SEQ ID NO: 2 in the sequence listing; miRNA avi- miR-316 antogimir has a single-stranded structure, and its nucleotide sequence is shown in SEQ ID NO: 3 in the sequence listing.

[0027] In order to better characterize the effects of miRNA avi-miR-316 agomir and miRNA avi-miR-316antagomir provided by the present invention, the present invention also designed negative controls agomir NC and antagomir NC, wherein the nucleosides of the two chains of agomir NC The acid sequences are respectively shown in SEQ ID NO: 4 in the sequence lis...

Embodiment 2

[0033] Example 2: Using miRNA avi-miR-316 agomir and miRNA avi-miR-316 antagomir to feed aphids and the lethal effect

[0034] 1. Preparation method of miRNA avi-miR-316 agomir, miRNA avi-miR-316 antagomi, agomir NC and antagomir NC:

[0035] (1) Use ultrapure water to prepare a sucrose solution with a mass fraction of 30%, then filter and sterilize it with a 0.2 μm microporous membrane, divide the obtained filtrate into 1.5ml sterilized centrifuge tubes, and store in a -20°C refrigerator spare;

[0036] (2) Dilute the synthesized and modified miRNA avi-miR-316agomir, miRNA avi-miR-316 antagomir, agomir NC and antagomir NC to a concentration of 300nM with a 30% sucrose solution to obtain miRNA avi-miR -316 agomir (+ sucrose solution), miRNA avi-miR-316 antagomir (+ sucrose solution), agomir NC (+ sucrose solution) and antagomir NC (+ sucrose solution), set aside.

[0037] 2. miRNA avi-miR-316 agomir, miRNA avi-miR-316 antagomi, agomir NC, antagomir NC and NC feeding aphids ...

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Abstract

The invention relates to a miRNA molecule which is of a double chain structure. The miRNA molecule comprises a positive-sense strand and a negative-sense strand, wherein the nucleotide sequence of the positive-sense strand is as shown in SEQ ID NO: 1 in a sequence table, and the nucleotide sequence of the negative-sense strand is as shown in SEQ ID NO: 2 in a sequence table; or the miRNA molecule is of a single-strand structure, the nucleotide sequence of which is as shown in SEQ ID NO: 3. The designed and synthesized miRNA avi-miR-316 agomir and miRNA avi-miR-316 antagomir can make aphids fatal, thereby providing a novel effective product for prevention and control of aphids. A target gene acted thereby can be used as an ideal candidate gene resource for creating a novel anti-aphid crop medicated by RNAi, thereby providing a novel research concept for pest control. The miRNA molecule has very high research value and a wide application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an artificially designed and synthesized miRNA and its effect on controlling aphids. Background technique [0002] Wheat aphids are a worldwide agricultural pest, mainly including Sitobion avenae (Fabricius), Schizaphis graminum (Rondani), Rhopalosiphum padi (Linnaeus) Acyrthosiphon dirhodum (Walker) and others. Wheat aphids have the characteristics of wide distribution, large numbers, strong fecundity, and serious damage. They can migrate over long distances and spread viruses during the migration process. The honeydew excreted by them can also breed a variety of molds and affect wheat plants. photosynthesis and respiration, leading to crop yield reduction, causing huge losses to agricultural production, which has become one of the hot spots in the research of pest control in the agricultural field. At present, the control methods for aphids in China mainly use chemical pesticides...

Claims

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Application Information

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IPC IPC(8): C12N15/113A01N57/16A01P7/04
Inventor 李祥瑞朱勋张云慧魏长平张方梅程登发
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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