Migratory locust mesenchymal epidermal protein gene 6 and application thereof in locust control

A technology of epidermal proteins and internodes, applied in the fields of application, genetic engineering, and plant genetic improvement, can solve problems such as non-target biological hazards, environmental pollution, and drug resistance, and achieve the goal of reducing the number of eggs laid by pests and the number of offspring populations Effect

Inactive Publication Date: 2018-05-04
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the control of locust plague mainly relies on chemical control, but long-term application of chemical insecticides leads to a series of problems, such as environmental pollution, drug resistance and harm to non-target organisms, etc.

Method used

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  • Migratory locust mesenchymal epidermal protein gene 6 and application thereof in locust control
  • Migratory locust mesenchymal epidermal protein gene 6 and application thereof in locust control
  • Migratory locust mesenchymal epidermal protein gene 6 and application thereof in locust control

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Experimental program
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Embodiment 1

[0015] 1. Acquisition of the full-length sequence of the epidermal protein gene of the migratory locust intersegmental membrane

[0016] Based on the transcriptome database of migratory locusts, the epidermal protein gene of migratory locusts was searched using bioinformatics methods, and after sequence analysis combined with genome sequence splicing and comparison of migratory locusts, an intersegmental membrane epidermal protein gene of migratory locusts (LmAbd6 ) sequence, using primer premier5.0 software to design upstream and downstream primer sequences and send them to Sangon Bioengineering (Shanghai) Co., Ltd. for synthesis. Healthy adult migratory locusts of the same size were selected, and the intersegmental membrane tissue was dissected and frozen in liquid nitrogen. The first 6 biological replicates, 4 biological replicates were set up, and the total RNA was extracted according to the TaKaRa Trizol kit. Using M-MLV reverse transcriptase, the extracted total RNA was...

Embodiment 2

[0024] Embodiment 2: Experiment of killing migratory locusts by dsRNA of intersegmental membrane epidermal protein gene of migratory locusts

[0025] 1. Injection of dsRNA gene of migratory locust intersegmental membrane epidermal protein

[0026] Thirty female nymphs of the 5th instar and the 2nd day of age were selected for the experiment. 5 μl (10 μg) of dsRNA of SEQ ID NO: 3 and SEQ ID NO: 4 were gently injected with a 25 μl micro-syringe along the blood flow between the second and third abdominal segments of the lateral abdomen of the nymph. At the same time, 30 female nymphs were selected as the control group. Inject the same volume and concentration of dsGFP into the control group. Pick equal amounts of males that grow healthy and have the same size and mix with the injected female migratory locusts respectively and place them in a constant temperature biochemical incubator at 30°C to raise (light: dark time = 14h:10h, temperature 30±2°C, humidity 60% ), fed fresh wh...

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Abstract

The invention provides a migratory locust mesenchymal epidermal protein gene 6 and application thereof in locust control. The method specifically comprises the following steps: acquiring a migratory locust mesenchymal epidermal protein gene from a migratory locust transcriptome by virtue of a bioinformatics method, and further obtaining full-length genes having a sequence of SEQ ID NO:1 by virtueof a migratory locust genomic sequence. dsRNA of the gene is designed and synthesized according to the SEQ ID NO:1, the dsRNA is injected into a body cavity of the migratory locust, so that target genes can be specifically silenced, ecdysis of the body is difficult, the intersegmental membrane cannot be closed, and finally abdomens of injurious insects cannot deep into soil to lay eggs so as to greatly decrease the oocyst number and ovum number. Multiple experiments show that injection of the dsRNA causes reduction of the ovum number of offspring of the migratory locust by 70% or higher. Due to specificity and high efficiency of the gene disclosed by the invention, the gene has an obvious effect of reducing the number of laid eggs of the injurious insects and the offspring population quantity, and a novel pathway can be provided for pest control.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an application of migratory locust intersegmental membrane epidermal protein gene (LmAbd6) and its dsRNA in pest control. Background technique [0002] Migratory locust is an important agricultural pest in my country. Because it has wings and can migrate and fly between lands for a long distance, it has greatly improved its efficiency in foraging, courtship, avoiding enemies, and expanding distribution, causing serious harm to agricultural production. The body length of adult migratory locusts is 32.4-48.1 mm for males and 38.6-52.8 mm for females. Under normal circumstances, adults can mate about 7 days after eclosion, and females can mate many times throughout their lives, and can lay eggs under the soil 4 to 7 days after mating, at a depth of about 4 to 6 cm. When the eggs are laid, mucus is secreted to glue the eggs to form oocysts. Each female can generally lay 4 to 5 oocysts,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/113A01P7/04
CPCC07K14/43563C12N15/113C12N2310/14
Inventor 赵小明贾盼张建珍马恩波
Owner SHANXI UNIV
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