Application of oshox12 protein and its coding gene in improving rice resistance to sheath blight
The technology of encoding gene and transgenic rice is applied in the application field of OsHOX12 protein and its encoding gene in improving the resistance of rice to sheath blight, which can solve problems such as changes, and achieve the effect of improving resistance and reducing disease incidence.
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Embodiment 1
[0021] Example 1: Detection of relative expression levels of OsHOX12 at different time points after inoculation with sheath blight
[0022] Inoculate the sheath blight bacteria strain stored at 4°C—Rhizoctonia solani AG1-IA strain (the strain was provided by the laboratory of Wei Songhong, Shenyang Agricultural University) on the PDA medium in the ultra-clean workbench, and place it at 26°C The incubator was continuously cultured for 2-3 days, and the bacteria cake was taken from the edge of the colony with a 0.7cm sterile puncher for the inoculation experiment.
[0023] Put two sterilized filter papers soaked in sterile water into a sterilized petri dish, select healthy wild-type rice leaves of the same leaf age without damage by diseases and insect pests, and cut them into seven parts of equal size, with the back of the leaves facing the place neatly on the filter paper, use an inoculation needle or a toothpick to poke a small eye in the middle of each leaf, inoculate the fu...
Embodiment 2
[0032] Example 2: Obtaining of CRISPR / Cas9 Gene Editing Transgenic Rice Plant OsHOX12CR-KO
[0033] 1) Acquisition of transgenic rice
[0034] The OsHOX12 gene sequence (SEQ ID No.1) was provided to Baige Gene Technology Co., Ltd., and the company carried out target site sequence design, primer design, vector construction and transformation, and transformed rice variety Zhonghua 11. Finally, the T0 generation transgenic rice was obtained.
[0035] 2) Identification of transgenic rice
[0036] The obtained transgenic plants were further verified by sequencing analysis, and the results were as follows: figure 1 shown. Through the sequencing analysis of the positive plants, it was found that the base T was inserted after the 566th base of the OsHOX12 coding gene sequence shown in its sequence 1, so that the OsHOX12 gene function was lost ( figure 2 ), and named this gene-edited plant OsHOX12CR-KO.
Embodiment 3
[0037] Example 3: Phenotype observation of OsHOX12CR-KO transgenic rice after inoculation with sheath blight
[0038] Resistance identification was carried out by inoculating detached leaves of rice with sheath blight. The wild-type control and OsHOX12CR-KO transgenic rice leaves were inoculated with Rhizoctonia solani.
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