Transcription factor derived from arabidopsis thaliana and related to gray mold resistance and application thereof
A technology of Arabidopsis and Botrytis, applied in the fields of application, peptide source, genetic engineering, etc.
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Embodiment example 1
[0012] Implementation Case 1 Cloning of Arabidopsis C3H23 Gene and Acquisition of Overexpressed Plants
[0013] 1) Download the cDNA sequence of Arabidopsis C3H23 on the tair website, and design primers to amplify the fragment in the total cDNA of Arabidopsis after analyzing its specificity;
[0014] 2) After recovering and purifying the amplified fragment, construct it on an overexpression vector, extract the plasmid and transfer it into Agrobacterium after successful sequencing;
[0015] 3) Use Agrobacterium-mediated infection method of Arabidopsis inflorescence, and screen the transgenic positive plants on the resistance plate, extract transgenic plant RNA and reverse transcription, use qRT-PCR to identify transgenic plants with higher expression ;
Embodiment example 2
[0016] Implementation Case 2 Obtaining of Arabidopsis c3h23 Mutant Plants
[0017] After the purchased c3h23 mutants SALK-024606 and SALK-112158 were grown for four weeks, the DNA of the plants was extracted, PCR amplified and identified by agarose gel electrophoresis to obtain pure and mutants.
Embodiment example 3C3
[0018] Implementation case 3C3H23 gene in the pathogenicity of Botrytis cinerea
[0019] 1) Take the B. cinerea strain and culture it on the PDA medium, collect mature spores after 10 days of growth.
[0020] 2) After suspending the spores with liquid PDA, adjust the spore concentration to 5×10 5 CFU / mL, activate at 100rpm on a shaker at 25°C for 2h, select Arabidopsis wild-type, mutant and overexpression plants that have grown well for 4 weeks, and drop 6 μL of spore suspension onto three leaves at the same position of each plant, and cultivated in a humid environment.
[0021] 3) After 2 days, observe the phenotype and count the lesion area. The result is as figure 1 and figure 2 As shown, compared with the wild type, the c3h23 mutants SALK-024606 and SALK-112158 both showed larger lesion area after inoculation, while the lesion area of the overexpression plants was significantly reduced, indicating that the gene C3H23 has an increased effect on Botrytis cinerea. abi...
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