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

A technology of locust and insecticidal composition, applied in the field of ATP synthase β subunit gene, dsRNA and its application in the control of migratory locusts in East Asia, which can solve the problem of long killing time, high cost of control and unstable insecticidal effect and other issues, to achieve the effect of strong species specificity, strong gene specificity, and rapid onset of action

Active Publication Date: 2018-05-01
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 β subunit gene and its dsRNA application in pest control
  • An East Asian locust atp synthase β subunit gene and its dsRNA application in pest control
  • An East Asian locust atp synthase β subunit gene and its dsRNA application in pest control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

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

[0027] Based on the EST database of migratory locusts, the ATP synthase β subunit genes of migratory locusts were searched by bioinformatics methods.

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

[0029] Primers were designed based on the obtained nucleotide sequence of LmATP5B.

[0030] 3) Acquisition of total RNA from migratory locusts

[0031] Select 5th instar nymphs of Migratory locust with equal size, male and female, in groups of 3, and freeze them in liquid nitrogen for RNA extraction. Refer to the instruction manual of TRIzol (Invitrogen) for the specific operation steps.

[0032] 4) Synthesis of first-strand cDNA

[0033] The first-strand cDNA was synthesized accor...

Embodiment 2

[0043] Example 2: Five-instar migratory locusts killed by dsRNA of migratory locust ATP synthase β subunit

[0044] 1. DsRNA injection of migratory locust ATP synthase β subunit

[0045] 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 (SEQID NO: 3), and use a 25 μl micro-syringe for injection. Do not use too much force when injecting, and follow the direction of blood flow , the junction of the 2nd to 3rd abdominal segment of 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.

[0046] 2. Observation of the phenotype of the fifth instar migratory locust afte...

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PUM

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Abstract

The present invention provides the conserved sequence of cDNA of ATP synthase β subunit of locust, constructs its RNA, DNA and other constructs, utilizes RNAi technology to silence ATP synthase β subunit in locusts, and makes the ATP synthase β subunit in vivo 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 beta 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. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A01N57/16A01P7/04
Inventor 夏玉先胡军
Owner CHONGQING UNIV
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