Set of paddy rice anti-brown-planthopper genes, coded protein thereof, and application thereof
A technology against brown planthoppers and rice, applied in the field of genetic engineering, can solve the problems that the effect has not been reported yet
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Embodiment 1
[0042] Example 1. Discovery of rice resistance to brown planthopper-related proteins and their coding genes
[0043] 1. Resistance performance of rice variety Rathu Heenati to brown planthopper
[0044] Rice variety Rathu Heenati is highly resistant to brown planthopper. After being inoculated with N. lugens nymphs, Rathu Heenati showed normal plant growth and no obvious adverse symptoms. And the susceptible control 02428 showed curled leaves, and the plants wilted until dead (see figure 1 ).
[0045] 2. Anti-BPH gene mapping
[0046] 1. Preliminary mapping of the brown planthopper resistance gene
[0047] Detection of brown planthopper resistance genes using Rathu Heenati / 02428 hybrid F2 population. A main gene for resistance to BPH was detected in the interval of two SSR markers RM8213 and RM5953 on the short arm of chromosome 4, and named Bph17.
[0048] 2. Fine mapping of brown planthopper resistance genes
[0049] According to the results of preliminary mapping, u...
Embodiment 2
[0062] Embodiment 2, the acquisition and identification of transgenic plants
[0063] 1. Agrobacterium-mediated transformation
[0064] Using Agrobacterium strain EHA105 (purchased from Handsome Company, USA) as the mediator, the four gene recombination vectors constructed above were introduced into the susceptible species Kitaake.
[0065] (1) Cultivate Agrobacterium containing recombinant Bph17 for 16 hr at 28°C, collect the cells, and dilute them into N6 liquid medium containing 100 μmol / L to a concentration of OD 600 ≈0.5, to obtain the bacterial liquid;
[0066] (2) Mix and infect the mature embryogenic callus of rice that has been cultivated for one month with the above-mentioned bacterial solution for 30 minutes, blot the bacterial solution with filter paper, and then transfer to the co-cultivation medium (N6 solid co-cultivation medium, purchased by Sigma Company) medium, 24°C for 3 days;
[0067] (3) Inoculate the above callus on N6 solid selection medium containin...
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