Studying On Cloning And Function Of BolSDG8 Gene In Brassica Oleracea L

Flowering is an important step on the plant from vegetative growth to reproductive growth transformation, it paly an important role in plant all life. Arabidopsis SDG8 specifically regulates two and three methylation of H3K36 on the flowering key gene FLC, to promote its transcriptional expression and inhibit early flowering in plants. Brassica napus is allotetraploid fromed Brassica oleracea hybrid with Brassica campestris L, there are extensive cultivation in the Yangtze River region. Depth study to the mechanism of flowering in Brassica napus as an important role of rapeseed production. As compared with arabidopsis thaliana SDG8, we found a high similarity fragments with SDG8 in whole-genome sequence of Brassica oleracea. Brassica BolSDG8 gene were studied by bioinformatics, genetic engineering and molecular biology technical. We using four-step to explanation.1. As the Arabidopsis, Brassica oleracea is belong to Brassicaceae. Brassica oleracea BolSDG8 have high similarity with Arabidopsis SDG8. SDG8 and BolSDG8 are homologous gene, they all have an AWS Domain and a SET Domain. AWS domain is associated with SET domain, and its function is unclear. Post SET domain are abundant on histone lysine methyltransferase enzymes, which can provide an group of aromatic hydrophobic channel on histone C-terminal residues, and participation component activity sites. SET domain is evolutionarily conserved domains consist of 130 amino acids, which exists in almost all of the methyl transferase enzyme.2. By blasting BolSDG8 and SDG8, we chose a conserved sequence of 359bp named BolSDG8-RNA1. Via BolSDG8-RNA1 construction of RNA interference vector and genetic transformation into wild-type Columbia Arabidopsis plants, and obtained one transgenic plant with stable inheritance.3. Cloned BolSDG8 full-length gene into a expression vector, by genetic transformation into sdg8 Arabidopsis plants, and obtained one transgenic plant with stable inheritance.4. Through phenotype observation of two type transgenic plants in T3, and analysis of days to bolting and rosette leaves number, we found that BolSDG8 RNAi transgenic plants in Arabidopsis showed similar biological phenotypes to sdg8 mutants, weak and significantly early flowering for five days. The pFGC5941-BolSDG8-1 vector transformed into Arabidopsis sdg8 mutants, can conversion sdg8 mutant flowering becomes normal, further hint SDG8 and BolSDG8 has same biological function, laying a foundation for further study of biological function of BolSDG8.