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An ecdysone receptor gene ecr dsRNA and its application in controlling aphid damage

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: 2017-01-11
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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  • An ecdysone receptor gene ecr dsRNA and its application in controlling aphid damage
  • An ecdysone receptor gene ecr dsRNA and its application in controlling aphid damage
  • An ecdysone receptor gene ecr dsRNA and its application in controlling aphid damage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Obtaining of growth and development-related gene EcR dsRNA

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

[0028] About 20 tube aphids were taken respectively, 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 EcR genes of the pea aphid and green peach aphid (genbank accession numbers are NM_001159360 and EF174334, respectively, and the submission time is August 31, 2012 and April 17, 2007), primer P1 was designed using PrimerPrimer5.0 software (Table 1 ), which was synthesized by Beijing Huada Gene Company, was amplified and sequenced by using the cDNA of Echinochloa sativa as a template to obtain the molting-related gene EcR (SEQ ID NO:...

Embodiment 2

[0044] Embodiment 2, the application of dsRNA in inhibiting the growth of aphids

[0045] 1. Preparation of artificial diet for aphids and preparation of feeder

[0046] 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 , GaoL 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 The artificial feed was filtered with a bacterial filter, distributed into 2.0mL sterilized centrifuge tubes and stored in a -20°C refrigerator to avoid repeated freezing and thawing.

[0047] 2. Application of dsRNA (EcR dsRNA) in inhibiting the growth of aphids

[0048] Aphid feeding methods refer to the following documents: Jiao Min, Liu Shusheng. The technology of raising aphids with ...

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Abstract

The invention discloses an application of dsRNA of an ecdysone receptor (EcR) related gene 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 EcR gene 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 EcR dsRNA and its application in controlling aphid damage. Background technique [0002] Wheat aphids (especially aphids) are one of the main pests that harm wheat production in China. According to statistics, the area of ​​wheat aphid damage 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% reduction in production- 30%, 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-resista...

Claims

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Application Information

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