DsRNA of ecdysone receptor (EcR) gene USP and application thereof to control of damage caused by aphids

A technology of aphids and genes, applied in the direction of DNA/RNA fragments, applications, recombinant DNA technology, etc., can solve problems such as economic losses, lack of aphid-resistant germplasm resources of wheat, and difficulty in conventional breeding

Active Publication Date: 2014-10-22
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

[0004] There is a lack of aphid-resistant germplasm resources in wheat, and the resistance mechanism is still unclear. Conventional breeding is difficult to work, and it causes huge economic losses to agricultural production every year.

Method used

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  • DsRNA of ecdysone receptor (EcR) gene USP and application thereof to control of damage caused by aphids
  • DsRNA of ecdysone receptor (EcR) gene USP and application thereof to control of damage caused by aphids
  • DsRNA of ecdysone receptor (EcR) gene USP and application thereof to control of damage caused by aphids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1, the acquisition of growth and development-related gene USP dsRNA

[0027] 1. Extraction of total RNA and synthesis of cDNA

[0028] About 20 tube aphids were taken, and total RNA was extracted according to the instructions of the Trizol kit of Beijing Quanshijin Company, purified with RNA Cleaning Kit, and the first strand of cDNA was synthesized by reverse transcription. The operation steps refer to the instructions of the kit.

[0029] 2. Primer design and gene cloning

[0030] According to the conserved sequences of the USP genes of Aphid pisum and Myzus persica (genbank accession numbers are NM_001161668 and EF174335 respectively, and the submission time is March 10, 2011 and April 17, 2007), primer P1 was designed using Primer Primer5.0 software (Table 1), synthesized by Beijing Huada Genomics Co., Ltd., amplified and sequenced by using the cDNA of Aphid aphidica as a template to obtain the molt-related gene USP. The green fluorescent protein gene (GFP...

Embodiment 2

[0045] Embodiment 2, the application of dsRNA in inhibiting the growth of aphids

[0046] 1. Preparation of artificial diet for aphids and preparation of feeder

[0047] References for the preparation of artificial feed and feeder (Li Caixia, Gao Lifeng, Gao Lingling, Li Runzhi. Research on feeding aphids in pure artificial nutrient solution. Journal of Shanxi Agricultural University, 1997, 17(3): 225-228. Li C X , Gao L F, Gao L L, Li R Z. Study on the rearing of aphids on an artificially solidic diets. Journal of Shanxi Agricultural University, 1997,17(3):225-228.(in Chinese)), with 0.2μm Filter the artificial feed with a bacterial filter, dispense it into 2.0mL sterilized centrifuge tubes and store in a -20°C refrigerator to avoid repeated freezing and thawing.

[0048] 2. Application of dsRNA (USP dsRNA) in inhibiting the growth of aphids

[0049] Aphid feeding methods refer to the following documents: Jiao Min, Liu Shusheng. The technology of raising aphids with artific...

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Abstract

The invention discloses an application of dsRNA of an ecdysone receptor (EcR) related gene USP to control of damage caused by aphids. DsRNA provided by the invention is double-stranded RNA formed by a nucleotide shown in a sequence 2 in a sequence table and a nucleotide shown in an inverse complementary sequence of the sequence 2. Experiments prove that dsRNA of cDNA of the EcR related gene of sitobion avenae is obtained; expression of the gene USP in sitobion avenae can be inhibited, impaired growth and development of wheat aphids can be caused and lethal effects can be generated by adopting the method of feeding dsRNA in vitro.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an ecdysone receptor gene USP dsRNA and its application in controlling aphid damage. Background technique [0002] Wheat aphids are one of the main pests that harm wheat production in China. According to statistics, the area of ​​damage caused by wheat aphids in China can reach as high as 17 million hectares every year, accounting for 62% of the total wheat planting area, resulting in a 15%-30% reduction in production, and even up to 50% in severe cases. %. In recent years, due to factors such as global warming and changes in farming systems, the reproductive ability and adaptability of aphids have been significantly enhanced, and their harm has become increasingly serious. At present, aphid control is mainly based on spraying pesticides, but a large amount of pesticides are not only harmful to humans and animals, but also cause serious environmental pollution. Breeding aphid-resis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/11A01N57/16A01P7/04
Inventor 夏兰琴陈红梅孙永伟
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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