Coding gene OBP11 for OBPs (odorant binding proteins) of brown planthopper and application
A brown planthopper and brown planthopper-resistant technology is applied in the fields of application, genetic engineering, plant gene improvement, etc., to achieve the effect of reducing the amount of pesticides and manpower input
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Embodiment 1
[0021] The coding gene OBP11 of a brown planthopper odor binding protein is obtained by the following method:
[0022] Using the brown planthopper cDNA as a template, design primers O1: 5'-ATGAAGTCGTTCATCGTTTG-3'; O2: -TCACGGCCGTCCTCCTTCTTA-3' for PCR amplification. The PCR conditions are as follows:
[0023] 3min, 95℃; 30s 94℃; and 60s 57℃, 1min 72℃35 cycles; 10min, 72℃; 1min, 25℃.
[0024] Finally, the OBP11 gene was obtained, its sequence is shown in SEQ ID NO.1, and the corresponding amino acid sequence is shown in SEQ ID NO.2.
Embodiment 2
[0026] The application of OBP11 gene in the control of brown planthopper to breed new resistant rice varieties is as follows:
[0027] Using the rice 35S promoter and DS1301 as the interference vector, construct a recombinant vector capable of producing double-stranded RNA hairpin structure, and transform the constructed recombinant vector into Agrobacterium tumefaciens strain (EHA105). The PCR primers during the vector construction process are as follows : Upstream primer: 5'-CCGGAATTCATGAAGTCGTTC-3'; Downstream primer: -CCGCTCGAGTCACGGCCGTC-3', using the commonly used Agrobacterium-mediated genetic transformation method in this field, the embryogenic callus induced by Japonica rice Zhonghua 11 seeds was used as the recipient Soak and infect the callus with the Agrobacterium suspension medium containing the final carrier of the target gene, and select positive plants on the selection medium containing hygromycin. The specific method refers to the highly efficient genetic tran...
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