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Oriental migratory locust ATP synthase alpha subunit gene and application of dsRNA of oriental migratory locust ATP synthase alpha subunit gene in pest control

A technology of constructs and nucleic acid molecules, 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., and achieve strong gene specificity, species It is a highly specific and fast-acting effect

Active Publication Date: 2015-03-25
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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  • Oriental migratory locust ATP synthase alpha subunit gene and application of dsRNA of oriental migratory locust ATP synthase alpha subunit gene in pest control
  • Oriental migratory locust ATP synthase alpha subunit gene and application of dsRNA of oriental migratory locust ATP synthase alpha subunit gene in pest control
  • Oriental migratory locust ATP synthase alpha subunit gene and application of dsRNA of oriental migratory locust ATP synthase alpha subunit gene in pest control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

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

[0026] Based on the EST database of migratory locust, the ATP synthase α subunit gene of migratory locust was searched by bioinformatics method. After sequence splicing and comparison, a total of 1 ATP synthase α subunit gene of migratory locust was obtained.

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

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

[0029] 3) Acquisition of total RNA from migratory locusts

[0030] Select 5th instar nymphs of Migratory locust with equal size, male and female, in groups of 3, and freeze them in liquid nitrog...

Embodiment 2

[0044] Example 2: Five-instar migratory locusts killed by dsRNA of migratory locust ATP synthase α subunit

[0045] 1. dsRNA injection of migratory locust ATP synthase α subunit

[0046] Select nymphs on the third day of the fifth instar, uniform in size, and in the same health status to inject the above-mentioned synthetic dsRNA (SEQ ID NO: 3), and use a 25 μl micro-syringe for injection. Do not use too much force when injecting, and follow the blood flow. Direction, the junction of the 2nd to 3rd abdominal segment in the flank was used as the injection point, avoiding the valve. The amount of dsRNA injected was 100ng, and a dsGFP (100ng) control group was set up, with 20 worms in each group and 3 biological repetitions, a total of 60 worms. 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 ...

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Abstract

The invention relates to an oriental migratory locust ATP synthase alpha subunit gene and application of dsRNA of the oriental migratory locust ATP synthase alpha subunit gene in pest control. The oriental migratory locust ATP synthase alpha subunit gene disclosed by the invention provides the conserved sequence of ATP synthetase alpha subunit cDNA of a locust, can be used for constructing the constructs, namely RNA, DNA and the like, and obviously inhibits the expression of the ATP synthetase alpha subunit inside a locust body by silencing the ATP synthetase alpha subunit inside the locust body by utilizing an RNAi technology to cause the lethal effect on the locust, thus being applied to the RNAi technology to prevent and control oriental migratory locust pests.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an ATP synthase α subunit gene, dsRNA and its application in the control of East Asian migratory locusts. Background technique [0002] Pests seriously endanger agricultural production. Due to the long-term application of organochlorine, organophosphorus and other chemical insecticides, 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 insecticides, and the cost of control has increased. At present, biopesticides have been applied to pest control, but the killing time is long and the insecticidal effect is unstable. Therefore, new pest control methods are urgently needed. [0003] RNA interference (RNAi), a phenomenon of specific post-transcriptional gene silencing usually caused by double-stranded RNA molecules, was awarded the Nobel Prize in 2006. Th...

Claims

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Application Information

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