Identifying Cytoplasmic Male Sterile Types of Chinese Cabbage （Brassica campestris L. ） by Molecular Markers

摘要

Different sterile cytoplasm types of nine cabbage cytoplasmic male sterile materials were identified by molecular marker in the study,in order to better use molecular marker to conduct the assisted breeding in the future. Genomic DNA was isolated from Chinese cabbage by CTAB method. The design of two pairs of specific primers was performed on conserved flanking region of orf138 gene in the GenB ank. PCR was performed with genomic DNA of the nine Chinese cabbage materials. The bands were sequenced. The homologous comparison was conducted in NCBI,and finally,the type of sterile cytoplasm was determined. The results showed that the bands were amplified only in four Chinese cabbage male sterile materials with two pairs of specific primers PI/PII and PIII/PIV,while the other five materials did not obtain the relative bands. The result was consistent with the field sterility identification. And then four molecular markers of Chinese cabbage Ogura cytoplasmic male sterility( CMS) were obtained. After conducting a homologous comparative analysis with BLAST in GenB ank,it was found that the homologous degree was 100% in specific segments of the three sterility materials( L1-CI,L3-CI and L3-F1) and Ogu orf138 gene( GenB ank accession No. : HQ149728) of the reported broccoli Ogu CMS. The homologous degree of L1-F1 was 99% with a variation point. The type of cytoplasmic male sterility of the other five materials needed further research. Four materials of the nine were identified as the radish cytoplasmic male sterility materials and four molecular markers were obtained.