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The lethal gene csu-mir-9b of Chilo suppressalis and its application

A csu-mir-9b, 1.csu-mir-9b technology, applied in the field of agricultural biology, can solve environmental pollution and other problems, achieve the effects of low synthesis cost, resistance to degradation by RNase, and convenient use in a large number of experiments

Active Publication Date: 2018-06-29
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In my country, the continuous single long-term use of chemical agents has led to the resistance of Chilo borer to various pesticides to varying degrees. It is necessary to continuously increase the amount of pesticides used to achieve satisfactory control effects, resulting in more serious environmental pollution. vicious circle

Method used

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  • The lethal gene csu-mir-9b of Chilo suppressalis and its application
  • The lethal gene csu-mir-9b of Chilo suppressalis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) Chemically synthesize the lethal gene csu-miR-9b of Chipotle borer, the sequence is 5'-UCUUUGGUUAUCUAGCUGUAUGA-3';

[0026] (2) Purify using high performance liquid chromatography (HPLC);

[0027] (3) Use a spectrophotometer to accurately quantify, and freeze-dry for storage.

Embodiment 2

[0029] A lethal gene csu-miR-9b injection method of Chilo suppressalis comprises the following steps:

[0030] (1) diluting the csu-miR-9b described in claim 1 with nuclease-free water to a concentration of 200pmol / μL;

[0031] (2) Select the larvae of Chilo suppressalis for an appropriate number of days, and inject 0.5 μL / head of the csu-miR-9b solution on the upper back or side with a micro-syringe;

[0032] (3) Pick up the worm body with a needle, wait for a period of time, when the csu-miR-9b has mixed into the blood cavity of the larvae, gently pull out the needle;

[0033] (4) Place the injected larvae in a 12-well plate, add an appropriate amount of rice seedlings, cover the lid, tie it up with a rubber band, and observe the phenotype every 6 hours. The results are as follows: figure 1 As shown, it can be seen that the injection of csu-miR-9b has an obvious lethal effect on C. suppressalis, indicating that the miRNA gene can be used as an effective interferor of import...

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Abstract

The invention discloses the lethal gene csu-miR-9b of Chilo suppressalis and its application. The lethal gene csu-miR-9b of C. suppressalis borer, the sequence is 5'-UCUUUGGUUAUCUAGCUGUAUGA-3'. The application of the csu-miR-9b of the present invention in the preparation of an insecticidal agent for Chilo suppressalis. The present invention uses complete miRNA bioinformatics analysis to search for an effective miRNA—csu-miR-9b from a large amount of small RNA library data. Injection of C. suppressalis with csu-miR-9b synthesized in vitro has obvious lethal effect on C. suppressalis, indicating that the miRNA gene can be used as an effective interferer of important genes in C. suppressalis.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and relates to the lethal gene csu-miR-9b of Chilo suppressalis and its application. Background technique [0002] Chilo suppressalis (Walker), which belongs to Lepidoptera and Mothidae, is one of the most serious frequent pests of rice in my country. In my country, the continuous single long-term use of chemical agents has led to the resistance of Chilo borer to various pesticides to varying degrees. It is necessary to continuously increase the amount of pesticides used to achieve satisfactory control effects, resulting in more serious environmental pollution. vicious circle. Therefore, in the practice of agricultural production, there is an urgent need for alternative control methods other than chemical pesticides. [0003] microRNAs (miRNAs) are small, siRNA-like molecules that are encoded by the genomes of higher eukaryotes. miRNAs degrade mRNAs or hinder their translation by base-p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A01N57/16A01P7/04
Inventor 李飞葛畅王书平肖花美孙杨
Owner NANJING AGRICULTURAL UNIVERSITY
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