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Chilo suppressalis lethal gene csu-miR-260 and application thereof

A technology of csu-mir-260 and diploid borer, which is applied in the field of agricultural biology, can solve problems such as environmental pollution, and achieves the effects of low synthesis cost, resistance to RNase degradation, and convenient use in a large number of experiments

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-260 and application thereof
  • Chilo suppressalis lethal gene csu-miR-260 and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0026] (1) Chemically synthesize the lethal gene csu-miR-260 of Chilo suppressalis, the sequence of which is 5'-UUUUGGAUGACUGGCCCAUGUCGGCGU-3' (SEQ ID NO.1);

[0027] (2) Purify using high performance liquid chromatography (HPLC);

[0028] (3) Use a spectrophotometer to accurately quantify, and freeze-dry for storage.

Embodiment 2

[0030] A lethal gene csu-miR-260 injection method of Chilo suppressalis comprises the following steps:

[0031] (1) Dilute the csu-miR-260 to a concentration of 200 pmol / μL with nuclease-free water;

[0032] (2) Select the larvae of Chilo suppressalis for a suitable number of days, and inject 0.5 μL / head of csu-miR-260 solution on the upper back or side with a micro-syringe;

[0033] (3) Pick up the worm body with a needle, wait for a period of time, until the csu-miR-260 has mixed into the blood cavity of the larvae, and gently pull out the needle;

[0034] (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-260 has an obvious lethal effect on C. suppressalis, indicating that the miRNA gene can be used as an effective interferer of important genes in C. supp...

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Abstract

The invention discloses a chilo suppressalis lethal gene csu-miR-260 and application thereof. The sequence of the chilo suppressalis lethal gene csu-miR-260 is shown as 5'-UUUUGGAUGACUGGCCCAUGUCGGCGU-3'. The invention discloses application of the csu-miR-260 in preparing a chilo suppressalis insecticidal reagent. On the basis of complete miRNA bioinformatics analysis, an effective miRNA, namely csu-miR-260, is searched from mass data in a small RNA library. The csu-miR-260 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 lethal gene csu-miR-260 of Chilo borer and its application. Background technique [0002] Chilosuppressalis (Walker) belongs to Lepidoptera, Mothidae, and is one of the most serious frequent pests on 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-pairing with t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A01N57/16A01P7/04
Inventor 李飞王书平葛畅孙杨肖花美
Owner NANJING AGRICULTURAL UNIVERSITY
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