Yunnan Oryza meyeriana Baill MeXB3 gene and application thereof
The invention relates to a technology of wild rice verrucas and wild rice, which is applied in the field of molecular biology and can solve the problems such as public reports on the research of XB3 gene of wild rice verrucas, and achieve the effect of improving the resistance to bacterial blight of rice and enhancing the resistance.
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Embodiment 1
[0018] Example 1 Cloning and analysis of the gene MeXB3 related to resistance to bacterial blight of wild rice verruca in Yunnan
[0019] (1) Cultivation and processing of materials
[0020] Yunnan verrucous wild rice (Oryza meyeriana Baill) was grown in a greenhouse until flowering. Use NA medium to activate bacterial blight pathogen Y8, culture at 28°C for 2-3 days, wash the bacterial lawn with sterile distilled water, and prepare OD 600 0.6 bacteria solution. When the temperature in the greenhouse rose to 28°C, the leaves of O. yunnanensis were inoculated by the leaf-cut method. The control group was simulated inoculated with sterile water. After inoculation, samples were taken in equal quantities at 2h, 4h, 8h, 24h, 48h, and 72h. That is to say, the same amount of wild rice verruca yunnanensis was sampled at the same time, and the samples were quickly frozen in liquid nitrogen immediately after sampling, and stored in a -80°C refrigerator for later use.
[0021] (2) Ext...
Embodiment 2
[0033] Example 2 The application of the MeXB3 gene of Oryza verruculata to enhance the resistance to rice bacterial blight.
[0034] (1) Expression vector construction of MeXB3 gene
[0035] According to the full-length cDNA sequence of the bacterial blight resistance gene MeXB3 of wild rice verruca yunnanensis, primers containing the complete open reading frame were designed, and restriction endonuclease NcoI and BaglⅡ restriction enzyme cutting sites were added during the design to recombine the MeXB3 gene into the expression vector pCAMBIA2301. The base sequence of the upstream primer MeXB3-OF of the ORF sequence of MeXB3 is shown in SEQ ID NO.8 in the sequence listing, and the base sequence of the downstream primer MeXB3-OR of the ORF sequence of MeXB3 is shown in SEQ ID NO.9 in the sequence listing.
[0036] (2) Transformation of Agrobacterium
[0037] Take the competent LBA4404 Agrobacterium tumefaciens stored at -80°C and thaw on ice. Take 1 μL of the expression vect...
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