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
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[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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