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Chilo suppressalis lethal gene csu-miR-9b and application thereof

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

Active Publication Date: 2016-01-13
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Chilo suppressalis lethal gene csu-miR-9b and application thereof
  • Chilo suppressalis lethal gene csu-miR-9b and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) Chemical synthesis of the Chilo suppressalis lethal gene csu-miR-9b, the sequence is 5’-UCUUUGGUUAUCUAGCUGUAUGA-3’;

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

[0027] (3) Use a spectrophotometer for accurate quantification, freeze-dry storage.

Embodiment 2

[0029] A method for injecting the lethal gene csu-miR-9b of Chilo suppressalis includes the following steps:

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

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

[0032] (3) Use a needle to pick up the worm body, wait for a while, when csu-miR-9b has mixed into the blood cavity of the larva, gently pull out the needle;

[0033] (4) Place the injected larvae in a 12-well plate, add an appropriate amount of rice seedlings, cover 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 csu-miR-9b injection of Chilo suppressalis has a significant lethal effect on Chilo suppressalis, indicating that the miRNA gene can be used as an effective interference of important genes of Chilo suppressalis.

[0034]

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Abstract

The invention discloses a chilo suppressalis lethal gene csu-miR-9b and application thereof. The sequence of the chilo suppressalis lethal gene csu-miR-9b is shown as 5'-UCUUUGGUUAUCUAGCUGUAUGA-3'. The invention discloses application of the csu-miR-9b in preparing a chilo suppressalis insecticidal reagent. On the basis of complete miRNA bioinformatics analysis, an effective miRNA, namely csu-miR-9b, is searched from mass data in a small RNA library. The csu-miR-9b synthesized in vitro, injected to chilo suppressalis, has a significant lethal effect on the chilo suppressalis, showing that the miRNA can serve as an effective interferent for important genes of the chilo suppressalis.

Description

Technical field [0001] The invention belongs to the field of agricultural biotechnology, and relates to the chilo suppressalis lethal gene csu-miR-9b and its application. Background technique [0002] Chilosuppressalis (Walker) belongs to the order Lepidoptera and Mothidae. It is one of the most serious common pests on rice in my country. In my country, the continuous single and long-term use of chemical agents has caused the Chilo suppressalis to have varying degrees of resistance to a variety of pesticides. It is necessary to continuously increase the use of pesticides to achieve satisfactory control effects, resulting in more serious environmental pollution. Vicious circle. Therefore, in agricultural production practice, 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. The miRNA guides the silencing complex (RISC) to degrade mR...

Claims

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Application Information

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Patent Type & Authority Applications(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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