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Application of Locusta migratoria ATP synthase beta subunit gene and its dsRNA in pest control

A technology of constructs and nucleic acid molecules, applied in the direction of DNA / RNA fragments, applications, recombinant DNA technology, etc., can solve the problems of long killing time, high cost of control, unstable insecticidal effect, etc., and achieve strong species specificity , strong gene specificity, fast onset effect

Active Publication Date: 2015-03-11
CHONGQING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Pests seriously endanger agricultural production. Due to the long-term use of chemical pesticides such as organochlorine and organophosphorus, a series of problems have been caused: pesticide residues have caused serious environmental pollution; toxic pesticides have endangered human health; insects have become resistant to pesticides, and the cost of control has increased, etc.
At present, biopesticides have been applied to pest control, but the killing time is long and the insecticidal effect is unstable, so there is an urgent need for new pest control methods

Method used

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  • Application of Locusta migratoria ATP synthase beta subunit gene and its dsRNA in pest control
  • Application of Locusta migratoria ATP synthase beta subunit gene and its dsRNA in pest control
  • Application of Locusta migratoria ATP synthase beta subunit gene and its dsRNA in pest control

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1: Obtaining of migratory locust ATP synthase β subunit gene and its dsRNA

[0025] i. Acquisition of ATP synthase β subunit gene of migratory locust

[0026] 1) Searching of migratory locust ATP synthase β subunit gene in migratory locust EST database

[0027] Based on the EST database of migratory locusts, a bioinformatics method was used to search the migratory locust ATP synthase β subunit gene. After sequence splicing and alignment, a total of 1 migratory locust ATP synthase β subunit gene was obtained.

[0028] 2) Design of primers for migratory locust ATP synthase β subunit gene

[0029] Based on the obtained base sequence of LmATP5B, primers are designed.

[0030] 3) Obtaining total RNA of migratory locust

[0031] Select 5th instar nymphs of the same size, male and female half migratory locust, 3 heads in a group, frozen in liquid nitrogen, to be extracted RNA, the specific operation steps refer to TRIzol (Invitrogen) instructions.

[0032] 4) Synthesis of the first...

Embodiment 2

[0043] Example 2: Migratory locust ATP synthase β subunit dsRNA kills the fifth instar locust

[0044] 1. Injection of migraine ATP synthase β subunit dsRNA

[0045] Select nymphs of the third day of fifth instar, uniform size, and consistent health status to inject the above-mentioned synthetic dsRNA (SEQ ID NO: 3), and 25μl micro syringe for injection. Do not use excessive force during injection and follow the blood flow. In the direction, the junction of the second to third abdominal segments of the flanks is used as the injection point, avoiding the valve. The amount of dsRNA injected was 100ng, and a control group of dsGFP (100ng) was set up with 20 worms in each group, 3 biological replicates, and a total of 60. After the injection, the test insects were raised in plastic cages (light: dark time 14h:10h, temperature 30±2°C), and fresh wheat seedlings were given.

[0046] 2. Observation of the phenotype of the fifth instar migratory locust after injection of dsRNA

[0047] Afte...

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Abstract

The invention provides a conserved sequence of locust's ATP synthase beta subunit cDNA, and constructs the RNA, DNA and other constructs thereof. The RNAi technology is utilized to silence the ATP synthase beta subunit in locusts, so that expression of in vivo ATP synthase beta subunit can be significantly inhibited, thus resulting in the lethal effect of locusts. The Locusta migratoria ATP synthase beta subunit gene and its dsRNA can be applied to the RNAi technology for control of Locusta migratoria pests.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to an ATP synthase beta subunit gene, dsRNA and their application in the prevention and control of migratory locusts in Asia. Background technique [0002] Pests seriously harm agricultural production. The long-term application of chemical pesticides such as organochlorine and organophosphorus has caused a series of problems: pesticide residues cause serious environmental pollution; toxic pesticides endanger human health; insects become resistant to pesticides, and control costs increase. At present, biological insecticides have been applied to pest control, but the insecticidal time is long and the insecticidal effect is unstable. Therefore, a new type of pest control method is urgently needed. [0003] RNA interference (RNAi) is a specific post-transcriptional gene silencing usually caused by double-stranded RNA molecules. It won the Nobel Prize in 2006. This discovery not only provides a m...

Claims

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Application Information

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IPC IPC(8): C12N15/113A01N57/16A01P7/04
Inventor 夏玉先胡军
Owner CHONGQING UNIV
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