Molecular cloning and characterization of the genie male sterility related gene CYP86MF in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee)

摘要

The difference was compared using mRNA differential display PCR (DD-PCR) technology in the flower buds of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee) between the genie male-sterile line (A line) and the maintainer line (B line). Eighteen differential fragments were obtained in a total of 120 selective amplifications. A differential fragment named B0302-4 was confirmed by Northern hybridization. The fragment B0302-4 was differentially associated with a maintainer line, and expressed specially in flower buds. The gene sequences of 3 min end and 5 min end were identified by 3 min RACE and 5 min end PCR amplification. The full cDNA named as CYP86MF is composed of 1667 base pairs including a 1578 bp-long open reading frame, and has 85.3 percent nucleotide similarity to a gene CYP86C4 encoding cytochrome P450 from Arabidopsis, and has 84.9 percent amino acid similarity to CYP86C4. Phylogenetic analysis based on amino acid sequences alignment reveals that CYP86MF were very close to the CFP86C4 gene indicating that CYP86MF might belong to the genes of the CFP86 subfamily.