Application of p3 gene of rice stripe virus in preparation of transgenic bacterial-blight-resistant frond

A rice streak virus, anti-bacterial blight technology, applied in the application, plant regeneration, plant products and other directions, can solve the problems of ecological environment and human health, increase the use of pesticides, etc., and achieve rich anti-bacterial blight Disease genetic resources, the effect of improving resistance

Active Publication Date: 2014-01-08
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
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Problems solved by technology

The traditional control method is to control rice blast by constantly changing pesticide varieties and increasing the use of pesticides, which puts pressure on the ecological environment and human health

Method used

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  • Application of p3 gene of rice stripe virus in preparation of transgenic bacterial-blight-resistant frond
  • Application of p3 gene of rice stripe virus in preparation of transgenic bacterial-blight-resistant frond
  • Application of p3 gene of rice stripe virus in preparation of transgenic bacterial-blight-resistant frond

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0035] The rice variety transferred in the present invention is Nipponbare.

[0036] 1. Acquisition of Recombinant Agrobacterium

[0037] (1) p3 cloning and vector construction

[0038] According to the analysis of RSV p3 sequence in GenBank, the present invention designs the following primers for amplifying the p3 gene sequence from RSV-infected rice diseased plants collected in the field. Restriction sites have been added to the primer design to recombine the p3 gene into the binary expression vector pCV1300. The primer sequences are as follows:

[0039] P3 (+): 5’-T TCTAGA ATGAACGTGTTCACATCGTCTGT-3’

[0040] P3 (-): 5’-G GGATCC CTACAGCACA GCTGGAGAGCT -3'

[0041] (2) Transformation of Agrobacterium

[0042]Take the competent Agrobacterium EHA105 stored at -70 °C and thaw on ice. Take 1 μl of the extracted pure pCV-P3-C plasmid and add it to 100 μl of the competent medium, mix well, and add it to the treated electric shock cup. Set the voltage of 2200V, electric...

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Abstract

The invention relates to application of a p3 gene of a rice stripe virus in the preparation of a transgenic bacterial-blight-resistant frond. According to the application, the p3 gene of the rice stripe virus is utilized for constructing a binary expression vector to successfully transform rice so as to obtain a bacterial-blight-resistant transgenic plant. An experimental result shows that the p3 gene expresses in a regenerated frond and proves that a regenerated plant has very strong bacterial blight resistance. Currently, most bacterial-blight-resistant genes are rice endogenous genes; being a viral protein, the p3 gene improves the bacterial blight resistance of the rice, enriches the gene resources of the bacterial blight and has important theoretical and practical significances on cultivation of the rice with high bacterial blight resistance.

Description

technical field [0001] The invention relates to the technical field of genetic engineering and the field of plant disease control, in particular to rice stripe virus ( Rice stripe virus , RSV) P3 gene application field in plant bacterial blight resistance. Background technique [0002] Rice stripe virus ( Rice Stripe virus , RSV) is an important rice virus transmitted by the small planthopper. The rice stripe blight caused by this virus continues to occur in my country, causing serious economic losses. In recent years, Chinese scientists have carried out a series of meticulous and in-depth work on the biology and molecular biology of RSV: the whole genome sequence of the virus has been determined, the virus silencing suppressor protein and motor protein have been identified, and the rice interaction protein has been isolated. Screening work, establishment of SBPH feeding poison, detection technology, comprehensive control technology and other work, the progress made in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/34C12N15/84C12N1/21A01H4/00A01H5/00
Inventor 燕飞吴根土杨勇严成其陈剑平
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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