| Non-heading Chinese cabbage(Brassica capmestris ssp.chinensis Makino) originated in China, is a subspecies of cruciferous Brassica species (Brassica capmestris L.). In recent years,Non-heading Chinese cabbage has been extensive introduced and cultivated not only in northern China, but also in Southeast Asia, Japan, the United States and some European countries and so on, gradually becoming a world vegetable. In this study,on the one hand, comparative of the botanical characteristics of Non-heading Chinese cabbage new germplasm line P70-203and maintainer,on the other hand, molecular marker techniques identify non-Chinese cabbage male sterile lines P70-203cytoplasmic type, and identify hybrid generation purity.This study speeds up the process of Non-Chinese cabbage breeding,supplies molecular basis for production applications.1. Breeding process of Non-heading Chinese cabbage cytoplasmic male sterile line P70-203is through the backcross breeding of the accidental discovery of male-sterile plants.In this study, observations of their botanical characteristics results show that:Non-heading Chinese cabbage CMS P70-203plants regular and robust, performance the green colour but not the yellow,and showing stable sterility reached100%sterility degree; The floral which is well-developed has normal nectary and perfect functional pistil.;Auther is not observed in stamen; Nectaries has enough nectar to attract insects and other insects to pollination. So the male sterile lines is with fine botanical characters. This study lays the foundation for preparation more first generations hybrids, promotes the range of CMS P70-203applications.2. In this research, we analyzed the type of cytoplasmic male sterile line of non-heading Chinese cabbage, P70-203, and its maintainer, P60-27-1, by PCR approaches based on the specific primer pairs designed from the mitochondrial genes of Polima CMS and the Ogura CMS, which would be close related to the cytoplasmic male sterile characters. The results showed that the specific750bp length of fragments from the tested male sterile line was amplified by using Ogura primer pair P1/P2, nor from the maintaining lines as well. And also the primers of Polima P3/P4, P5/P6was in available for amplifying in the both of the tested male sterile line and its maintainer. Blastn analysis NCBI online indicated that the amplified750bp length DNA shares99%high homology with that of the mitochondrial gene of Ogura cytoplasmic male sterile line in Broccoli (accession number:EU604643), and the same degree of homology are found between the750bp segment and the mitochondrial gene of the Ogura cytoplasmic male sterile line in radish (accession number: AB055438). These results could reveal that the cytoplasmic male sterile of P70-203, a New Germplasm of Non-heading Chinese Cabbage, might belong to the type of Ogura cytoplasmic sterile.3. Fingerpints identify non-Chinese cabbage hybrid generation08-1-61and its parents through RAPD, SRAP, SSR markers. Results showed that:one RAPD primer and one pairs of SRAP primers amplify complementary characteristics bands of both parents in hybrid generation amplification map.So they can effectively identify parents and hybrids F1; Two RAPD primers and one pairs of SRAP primers are for the partial female type; One RAPD and one pairs of SRAP primers are for the partial parent type; No selected SSR primers showed polymorphism,through primer screening of160RAPD primers,121pairs of SRAP primers,87pairs of SSR. Comparative analysis of three kinds of markers, RAPD markers amplified band patterns are more abundant, lower cost and easy to operate, but poor repro-ducibility; SSR markers possess best the repeatability. but the lowest the polymorphism the high cost of primer development;SRAP marker amplification map is the most abundant bands, more polymorphism, good reproducibility, was the better means to identify hybrid seed purity. |
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