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Application of the kit containing miRNA molecules in the preparation of products for controlling aphids

A kit and molecular technology, applied in DNA/RNA fragments, applications, recombinant DNA technology, etc., can solve the problems of lack of aphid resistance genes, etc., and achieve high research value and broad application prospects

Active Publication Date: 2020-02-14
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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  • Application of the kit containing miRNA molecules in the preparation of products for controlling aphids
  • Application of the kit containing miRNA molecules in the preparation of products for controlling aphids
  • Application of the kit containing miRNA molecules in the preparation of products for controlling aphids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Design and synthesis of miRNA avi-miR-184-3p agomir and miRNA avi-miR-184-3p antagomir

[0026] The corresponding miRNA avi-miR-184-3p agomir (miRNA agonist) and miRNA avi-miR-184-3p antagomir (miRNA antagonist) were designed based on the mature sequence of aphid miRNA avi-miR-184-3p. The miRNA avi-miR-184-3p agomir has a double-stranded structure, and the nucleotide sequences of the two strands are respectively shown in SEQ ID NO: 1 in the sequence listing and SEQ ID NO: 2 in the sequence listing; the miRNA avi-miR-184-3p 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-184-3p agomir and miRNA avi-miR-184-3p antagomir provided by the present invention, the present invention also designed negative controls agomir NC and antagomir NC, wherein the two agomir NCs The nucleotide sequence of the single chain is shown in SEQ...

Embodiment 2

[0033] Embodiment 2: Using miRNA avi-miR-184-3p agomir and miRNA avi-miR-184-3pantagomir to feed aphids and the lethal effect caused

[0034] 1. Preparation method of miRNA avi-miR-184-3p agomir, miRNA avi-miR-184-3p 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 place in a -20°C refrigerator Save for later use;

[0036] (2) The synthesized and modified miRNA avi-miR-184-3pagomir, miRNA avi-miR-184-3p antagomir, agomir NC and antagomir NC were diluted to a concentration of 300nM using a 30% sucrose solution to obtain miRNA avi-miR-184-3p agomir (+ sucrose solution), miRNA avi-miR-184-3pantagomir (+ sucrose solution), agomir NC (+ sucrose solution) and antagomir NC (+ sucrose solution), set aside.

[0037] 2. miRNA avi-miR-184-3p agomir, miRNA avi-miR-184-3p ...

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Abstract

The invention relates to a miRNA molecule. The miRNA molecule is of a two-strand structure containing a sense strand and an antisense strand, wherein the nucleotide sequence of the sense strand is shown as SEQ ID NO: 1 in a sequence table, and the nucleotide sequence of the antisense strand is shown as SEQ ID NO: 2 in the sequence table; or the miRNA molecule is of a single-strand structure of which the nucleotide sequence is shown as SEQ ID NO: 3 in the sequence table. According to the artificially designed and synthesized miRNA avi-miR-184-3p agomir and miRNA avi-miR-184-3p antagomir have lethality for aphid, a novel effective product is provided to prevent and control aphid; the target gene under the action of the product can be used as an ideal candidate gene resource for developing RNAi-mediated novel pest resisting crops. Therefore, a novel research ideal on pest prevention and control is provided; the miRNA molecule is high in research value and has a good application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application and method of a kit containing miRNA molecules in the preparation of aphid control products. 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A01N57/16A01P7/04
CPCA01N57/16C12N15/113C12N2310/141
Inventor 李祥瑞张云慧魏长平张方梅朱勋程登发
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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