Arabidopsis thaliana BVD1 gene and its use
An Arabidopsis, gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem of not being able to accurately control the absorption amount and location of exogenous hormones
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Embodiment example 1
[0022] 1. Arabidopsis mutant bushy and dwarf 1 (bud1), the original wild-type material is Col-0 ecotype.
[0023] 2. Plant cultivation
[0024] Arabidopsis was cultivated with vermiculite poured with 1 / 3 B5 culture solution, the growth condition was continuous light at 23°C, and the intensity was 80-120μmol·m -2 sec -1 . The aseptic culture of Arabidopsis thaliana adopts surface sterilization. After 70% alcohol surface sterilization for 3-5 minutes, soak and sterilize with 10% Bleach for 10-15 minutes, and wash with sterile water for 3-4 times. Treat at 4°C for 4 days to promote growth after germination on 1 / 2 MS medium under the same conditions as vermiculite culture.
[0025] 3. Co-segregation experiment of bud1 and T-DNA
[0026] Genomic total DNA was extracted from 10 wild-type plants and 70 mutant plants from the F2 population of bud1 homozygous and wild-type Arabidopsis hybrids. Plant genomic DNA was extracted using the CTAB method created by Lijiayang in 1995. The...
Embodiment example 2
[0038] Agrobacterium tumefaciens-mediated transformation of Arabidopsis
[0039] The construction of the transformation plasmid overexpressing BUD1 was obtained by ligating the genomic PCR product of BUD1 between the BamH I and Sac I sites of pBI121, and the primers used were 5'-GGATCCTCTCTCTTCTATTTCCATGGC-3' and 5'-GAGCTCACAAGCAGTCGGATAAAG-3'. Transform the constructed transformation plasmid into the Agrobacterium GV3101 (pMP90) strain by electric shock method. After colony PCR identification, transform the Arabidopsis Col-0 wild type by the vacuum method established by Bechtold N1993. On the 1 / 2 MS medium of prime (50ng / l), when the T1 generation plants grow to 4-6 rosette leaves, move them into the soil, and if necessary, take a rosette leaf to extract genomic DNA for detection. After the single plant of the T1 generation was harvested, some seeds were screened with kanamycin to observe the segregation of the T2 generation, and some were sown in vermiculite to obtain stable...
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