An East Asian locust atp synthase alpha subunit gene and its application in pest control

A technology for locusts and migratory locusts, applied in the direction of DNA/RNA fragments, applications, genetic engineering, etc., can solve the problems of long killing time, high cost of control, unstable insecticidal effect, etc., to achieve strong species specificity, gene Strong specificity and fast onset of effect

Active Publication Date: 2017-07-11
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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  • An East Asian locust atp synthase alpha subunit gene and its application in pest control
  • An East Asian locust atp synthase alpha subunit gene and its application in pest control
  • An East Asian locust atp synthase alpha subunit gene and its application in pest control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Obtaining of migratory locust ATP synthase alpha subunit gene and its dsRNA

[0024] i. Acquisition of migratory locust ATP synthase alpha subunit gene

[0025] 1) Searching of migratory locust ATP synthase α subunit gene in migratory locust EST database

[0026] Based on the migratory locust EST database, a bioinformatics method was used to search the migratory locust ATP synthase alpha subunit gene. After sequence splicing and alignment, a total of 1 migratory locust ATP synthase alpha subunit gene was obtained.

[0027] 2) Design of primers for migratory locust ATP synthase α subunit gene

[0028] Based on the obtained base sequence of LmATPSB, primer premier5.0 software was used to design primers. All primers were synthesized by Nanjing Jinsirui Biological Co., Ltd.

[0029] 3) Obtaining total RNA of migratory locust

[0030] Select 5th instar nymphs of the same size male and female half migratory locusts, 3 heads in a group, frozen in liquid nitrogen, to be extract...

Embodiment 2

[0044] Example 2: Migratory locust ATP synthase alpha subunit dsRNA killed the fifth instar locust

[0045] 1. Injection of migratory locust ATP synthase alpha subunit dsRNA

[0046] Select the nymphs of the third day of fifth instar, uniform size, and consistent health status to inject the above-mentioned synthetic dsRNA (SEQID NO: 3), and 25μl micro syringe for injection. Do not use excessive force during injection and follow the direction of blood flow. , 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. Each group had 20 worms, 3 biological replicates, and a total of 60. After the injection, the test insects were reared in plastic cages (light:dark time 14h:10h, temperature 30±2°C), and fresh wheat seedlings were given.

[0047] 2. Observation of the phenotype of the fifth instar migratory locust after injection of dsRNA...

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PUM

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Abstract

The present invention provides the conserved sequence of cDNA of the ATP synthase α subunit of locust, constructs its RNA, DNA and other constructs, utilizes RNAi technology to silence the ATP synthase α subunit in the locust body, and makes the ATP synthase α subunit in the body The expression is obviously inhibited, leading to lethal effect on locusts, and can be applied to the control of East Asian migratory locust pests by RNAi technology.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to an ATP synthase alpha subunit gene, dsRNA and their application in the prevention and control of migratory locusts in Asia. Background technique [0002] Pests seriously endanger 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 increase control costs. 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 phenomenon of specific post-transcriptional gene silencing usually caused by double-stranded RNA molecules. It won the Nobel Prize in 2006. This discovery not...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/55C12N15/11C12N15/63A01N57/16A01P7/04
Inventor 夏玉先胡军
Owner CHONGQING UNIV
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