Molecular Markers Analysis Sterility Gene Of CMS451 And RGms03-632 In Cabbage

In this research, analysing sterile gene cytoplasmic male sterile of Oguar cabbage CMS451 and maintainer line B451, recessive male sterile line RGms03-632 by RAPD and SRAP ,obtained the reaction systems of RAPD and SRAP which were suitable to cabbage and three specific bands which linked to sterile gene in cabbage.The main results are as follow:1. The genomic DNA from the variety of cabbage was extracted with CTAB,SDS Urea,respectively.Results showed SDS was optimal genomic DNA extraction method.2. RAPD reaction system of cabbage was optimized,total volume was 25μL, including 2.0mmol/LMgCl2,0.2mmol/LdNTPs,0.56μmol/Lprimer,0.5uTaq,DNA20ng,2.5μL10×buffer the rest was supplied with ddH2O.3 SRAP reaction system of cabbage was optimized,total volume was25μL, including 2.4mmol/LMgCl2,0.3 mmol/LdNTPs, 0.1μmol/L primer, 1.5 uTaq, DNA30ng, 2.5μL 10×buffer,2.5μL10×buffer,the rest was supplied with ddH2O.4. Ogura CMS cabbage was studied By RAPD. Results showed that S20-495 was believed to be the specific band in sterile pool.Though NCBI ,S20-495 sequence has higher alignment score with the SRK-46,SP11-46,SLG-46 gene sequence up to78%.Recessive male sterile line RGms03-632 was studied By RAPD. Results showed that S1301-639 was believed to be the specific band in sterile pool. The Genetic distance was 8.854cM identified by Joinmap ver3.0 software.Though NCBI ,S1301-639 sequence has higher alignment score with with Brassica self-incompatibility line S-15, SP11A gene sequence up to 78%.5. Recessive male sterile line RGms03-632 was studied By SRAP. Results showed that a pair of primer Me6Em8 was believed to be the specific band in sterile pool,the band was between 250-500bp. The Genetic distance was 22.548cM identified by Joinmap ver3.0 software.