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Application of knockout or silencing odorant binding protein gene 1 of aethina tumida to prevention and treatment of aethina tumida

An odor-binding protein, honeycomb technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve non-target biological hazards, environmental pollution, drug resistance and other problems

Active Publication Date: 2021-12-07
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, its control 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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  • Application of knockout or silencing odorant binding protein gene 1 of aethina tumida to prevention and treatment of aethina tumida
  • Application of knockout or silencing odorant binding protein gene 1 of aethina tumida to prevention and treatment of aethina tumida

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1: Obtaining the sequence of the honeycomb beetle odor binding protein gene 1 (OBP1) and its dsRNA

[0015] 1. Acquisition of the sequence of Odor Binding Protein Gene 1 (OBP1) of the hive beetle

[0016] 1) Search of the hive beetle OBP1 gene sequence in the hive beetle transcriptome database

[0017] Based on the hive beetle transcriptome database, bioinformatics methods were used to search the sequence of the OBP1 gene of the hive beetle. After sequence analysis and comparison, a total of one OBP1 gene sequence of the hive beetle was obtained.

[0018] 2) Design of primers required for PCR amplification:

[0019] Based on the above gene fragments, the upstream primer GTAAAAATGAAAACATTCGT and the downstream primer CGGCTAAATGAGCAGCCGTCAT were designed by Primer Premier 5.0 software and synthesized by Shanghai Sangon Bioengineering Co., Ltd. Total RNA was extracted from the antennae of adults of the honeycomb beetle on the seventh day, and the extracted RNA...

Embodiment 2

[0029] Example 2: Injection of dsRNA of Odor Binding Protein Gene 1 (OBP1) achieves a significant decrease in the expression level of OBP1 in the head of the beetle beetle

[0030] 1. Specific dsRNA injection

[0031] 将1μL(1μg)由SEQ ID NO:2所示的dsRNA(dsOBP1,具体为ACTCACAAATGCCGTGTCCAGAGAATTTGTGGAAAATTTTATGAATAAACTACAGGAAGTTGGGGAAAAGTGCGCAGAGGAAACCAATGCAACTGACGACGACGTAGCTGCATTAATTGCCCATACGATGCCCGAATCCCACAATGGAAAATGTATGATTTTGTGTTTTAATGTTGCTTTTCATTTGCAAAAACCTGATGGCACCCCAGATAAAGAAG)用10μL规格微量注射器注射到蜂巢小甲虫第七日龄幼虫体内,共注射100只。 注射相同体积浓度的dsGFP(GTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCTCTTATGGTGTTCAATGCTTTTCAAGATACCCAGATCATATGAAACAGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTATAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACA...

Embodiment 3

[0034] Example 3: Inhibiting the expression of the hive beetle OBP1, significantly reducing the response behavior of the hive beetle to pollen volatiles

[0035] Both the control group (injected with dsGFP) and the treatment group (injected with dsOBP1) were reared in an artificial climate chamber. After the 7th day of adult development, they were placed in a Y-shaped glass tube (direct 0.5 cm, main arm length 8 cm, branch arm length 5 cm) Olfactory behavioral experiments. Place a filter paper piece (1cm×1cm) at the end of the branch arm, add 5 μL n-hexane (control) or 5 μL n-hexane diluted pollen volatiles including ethyl linolenate (1 mg / mL) or ethyl palmitate dropwise on the filter paper piece (0.1 mg / mL). The experiment was carried out at an indoor temperature of 25°C, and an air pump was used to allow the airflow to pass through the activated carbon, humidification bottle, flow meter and filter paper successively at a constant speed of 0.8L / min. Put a single hive beetle...

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Abstract

The invention discloses an application of a knockout or silencing odorant binding protein gene 1 of aethina tumida to prevention and treatment of the aethina tumida. The nucleotide sequence of the odorant binding protein gene 1 of the aethina tumida is as shown in SEQ ID NO. 1. A RNA interference technology is adopted, dsRNA is used for silencing the OBP1 gene of the aethina tumida, selection of the aethina tumida on food source pollen volatile matter can be affected, the protein gene 1 can be used for preventing and treating the aethina tumida, and plays an important role in the prevention and treatment process of the aethina tumida.

Description

technical field [0001] The invention belongs to the field of insect control, and in particular relates to the application of knocking out or silencing the honeycomb beetle odor-binding protein gene 1 in preventing and controlling the honeycomb beetle. Background technique [0002] The hive beetle is a pest that parasitizes bee colonies. It was originally distributed in southern Africa, and then spread to Australia, America, Asia and other states and countries, causing various degrees of bee colony damage and economic losses all over the world. Hive beetles cause serious harm to bees in the larval and adult stages. They feed on pollen, honey and bee larvae, which can lead to the death of bee larvae, honey fermentation, and damage to the comb, often causing the entire hive to collapse and bee colonies to abandon the nest fly away. At present, its control mainly relies on chemical control, but long-term application of chemical insecticides leads to a series of problems, such a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/89C12N15/12A01K67/033
CPCC12N15/89C07K14/43563A01K67/0339A01K2227/706
Inventor 赵红霞武丽仙刘芳李良斌
Owner INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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