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

A rice streak virus, rice blast resistance technology, applied in the application, plant products, genetic engineering and other directions, can solve the problems of increasing the use of pesticides, the pressure on the ecological environment and human health, etc., to enrich the rice blast resistance gene resources, The effect of increasing resistance

Active Publication Date: 2014-01-08
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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

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Experimental program
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Embodiment Construction

[0037] The rice variety transferred in the present invention is Nipponbare.

[0038] 1. Acquisition of Recombinant Agrobacterium

[0039] (1) p3 cloning and vector construction

[0040] 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:

[0041] P3 (+): 5’-T TCTAGA ATGAACGTGTTCACATCGTCTGT-3’

[0042] P3 (-): 5’-G GGATCC CTACAGCACAGCTGGAGAGCT-3'.

[0043] (2) Transformation of Agrobacterium

[0044] 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 s...

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Abstract

The invention relates to application of a p3 gene of a rice stripe virus (RSV) in the preparation of a transgenic rice-blast-resistant frond. The application is characterized in that the nucleotide sequence of the gene is presented by SEQ ID NO:1; the gene is derived from the rice stripe virus and is produced by amplifying a sequence on a diseased rice strain collected from a farm by comparing and designing a primer through a GenBank sequence. Preferably, the gene is used for preparing rice-blast-resistant rice. The application has the advantages that (1) currently, most rice-blast-resistant genes are rice endogenous genes, and being a viral protein, the p3 gene can improve the bacterial blight resistance of the rice and enrich the rice-blast-resistant gene resources; (2) as the p3 gene is not a rice endogenous resistance gene, the resistance mechanism of the p3 gene is different from the resistance mechanisms of the other resistance genes and can help people to understand rice-blast-resistant bacteria mechanisms and rice blast bacteria pathogenesis of the rice.

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 , Application field of RSV) P3 gene in plant blast 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 these work...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 燕飞吴根土王教瑜王艳丽孙国昌陈剑平
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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