A powdery mildew resistance-related protein, its coding gene and its application
An encoding gene, powdery mildew technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve problems such as plant yield reduction, withering, plant growth and development effects, etc., to achieve the effect of variety improvement and pathogen resistance improvement
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Embodiment 1
[0031] Example 1, Preparation of powdery mildew resistance-related gene AT5G05190
[0032] (1) Screening of mutant Arabidopsis thaliana with enhanced resistance to powdery mildew
[0033] The wild-type Arabidopsis Col-0 was mutated with 0.03% ethyl methanesulfonate (EMS), and the mutants with enhanced resistance to powdery mildew were screened from the mutagenized population. The specific screening method is as follows: under a growth environment of 8000 lux and a relative humidity of 70% in short light (9 hours of light / 15 hours of darkness), use powdery mildew (G.cichoracearum UCSC1) (Adam et al., 1999) on the growth Four weeks of wild type and mutants were inoculated, and the experimental results were observed ( figure 1 ). The experimental results showed that: 7 days after inoculation, unlike the wild type that produced a large number of powdery mildew conidia, the hyphae and conidiophores could not grow on the surface of the leaves of the mutants, and the leaves of the ...
Embodiment 2
[0049] Example 2. Functional verification of powdery mildew resistance-related gene AT5G05190
[0050] (1), reverse mutation experiment
[0051] Digest the PCR amplification product obtained in Example 1 with Xba1 and Sal1; digest pCAMBIAL1300 with Xba1 and Sal1, and collect the large fragment of the vector; connect with T4 ligase at 16°C, transform DH10b competent cells, and find no base mutation by sequencing After extracting the plasmid (Sambrook, 1989), the exogenous gene sequence inserted in the plasmid is shown in SEQ ID No.1, and then the freeze-thaw method is used to transform the Agrobacterium competent GV3101, and then the Agrobacterium is used for flower drop dipping (Clough and Bent, 1998) transformed the mutants screened in Example 1, and screened transgenic plants with hygromycin to obtain 25 positive plants. These 25 positive plants were inoculated with powdery mildew, and 22 transgenic plants reverted to wild-type phenotype ( figure 2 b).
[0052] (2), AT5G...
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