Application of rice osmpkk10-2 gene in improving rice disease resistance and drought resistance
A gene, rice technology, applied in the field of plant genetic engineering, can solve the problems of food security situation impact, yield and quality decline, etc.
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Embodiment 1
[0023] Example 1: Analysis of the expression pattern of the OsMPKK10-2 gene after biotic and abiotic stress treatments
[0024] It has been reported that some mitogen-activated protein kinase genes in plants are involved in the regulation of plant disease resistance and / or drought resistance (Ning et al., 2010; Meng and Zhang, 2013). Benzothiadiazol (BTH) is a functional analog of the plant hormone salicylic acid (SA), which can activate the plant's disease resistance signaling pathway when exogenously applied to enhance plant disease resistance ( et al., 1996). ABA plays an important role in plant resistance to drought, high salinity and other stress responses (Zhu, 2002).
[0025] In order to verify whether the OsMPKK10-2 gene is involved in the regulation of rice resistance to disease and drought, the researchers of the present invention sprayed benzothiadiazole (BTH) or abscisic acid (ABA) on four-leaf stage japonica rice variety Zhonghua 11, and inoculated thin strips. ...
Embodiment 2
[0026] The test results showed that the expression of OsMPKK10-2 gene was rapidly induced after treatment with BTH and inoculation with S. finestripes; and the expression of OsMPKK10-2 gene was also rapidly induced after ABA treatment and drought stress ( figure 2 ). This result suggested that the OsMPKK10-2 gene may be involved in the regulation of rice resistance to thin stripe disease and drought stress, and enhancing the expression of this gene may improve the disease resistance and drought resistance of rice. Example 2: Determining the sequence and structure of the OsMPKK10-2 gene in rice variety Zhonghua 11
[0027] 1. Cloning of the sequence of the OsMPKK10-2 gene in the rice variety Zhonghua 11
[0028] According to the literature (Hamel et al., 2006), the LOC_Os03g12390 site in the rice genome sequence database (http: / / rice.plantbiology.msu.edu / ) contains the sequence information of the OsMPKK10-2 gene in the rice variety Nipponbare, Its corresponding cDNA sequence...
Embodiment 3
[0036] Example 3: Functional verification of OsMPKK10-2 gene
[0037] The invention adopts the genetic transformation method mediated by Agrobacterium to further verify the function of the gene by overexpressing the OsMPKK10-2 gene in the rice variety Zhonghua 11.
[0038] 1. Construction of genetic transformation vector
[0039] The starting vector used in the present invention is the commonly used vector pU1301 ( Figure 5 , Qiu et al., 2007). It is an Agrobacterium-mediated genetic transformation vector carrying a maize ubiquitin promoter with constitutive and overexpression characteristics.
[0040] Using the T-A cloning vector containing the full-length cDNA sequence of the OsMPKK10-2 gene in Example 2-2 as a template, use the PCR primer OsMPKK10-2-oe-F2 (5'-GG GGTACC TCCAACCGATGGCTCTC-3′) (the underline is the KpnI restriction site) and OsMPKK10-2-oe-R2 (5′-CG GGATCC TAGATCGCCGCTCTGCT-3') (the underline is the BamHI restriction site) to amplify the entire coding re...
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