| Asparagus (Asparagus officinalis L.), known as "the king of vegetable", have very high nutrition and economic value. In our country, the germplasm resources of asparagus is so deficiency that always depends on import. But there have another question, the price of import seed is too high and most of them are more fitting the arid zone. In the south of our country, especially in the hot area, disease of asparagus is prevalent. Phoma asparagi Sacc, as a fatal disease in the asparagus industry, the explore of disease-resistant cultivar is imperative.This paper have collected24asparagus germ plasm resources from domestic and overseas, the Inter-simple sequence repeats (ISSR) markers are provided to the study of genetic diversity; highly resistance resources identified by the pathogen of Phoma asparagi Sacc used to the material of tissue culture. and obtain the following results:1. three methods have been taken to extraction the genomic DNA of asparagus and explored the influence of the DNA quality by these methods:the improved SDS method, the improved CTAB method and the kit method. The results show that band is clear and tidiness extracted by the improved CTAB method. Also, DNA purity and yield is better than the other kinds of methods.2.A set of PCR efficient response analysis system has established to the ISSR analysis of asparagus germplasm resources.3.From the100ISSR primers screened10polymorphic primers for ISSR analysis of24varieties of asparagus. Screening of10primers for PCR amplification of24varieties of asparagus statistics to get98Amplification loci and74polymorphisms (SNPs), accounting for75.61%of the all amplified loci. The average number of each primer loci is10. Finally results by using NTSYS2.1software analysis showed that:the genetic similarity coefficient between0.76-0.93from the similarity coefficient of24asparagus germplasm is closer relationship. When the24asparagus germplasm in the similarity coefficient is0.771points,24species can be divided into four groups, and when the similarity coefficient is0.899points, the Dutch varieties all get together; the results of principal component analysis are consistent with the genetic similarity coefficient analysis.4.Apollo, Purple Passion,Grande and Jesse Knight are highly resistant varieties. These four varieties of disease resistance is the strongest. So these four varietes could be the object for the future study.5.Jersey knight varietiy of asparagus stem tip, stem section, seed as different explant induction on callus, and concludes that the stem tip is the best callus induction explant.6.Sterilization of Asparagus spears.-rapid disinfection of the moment with75%alcohol, then use sterile water washing for4-5times, the effective treatment is that treating by0.1%HgCl2for13min, the contamination rate decrease and survival rate rise up to100%.The most suitable variety for inducing callus of asparagus shoot tip, Jersey Knight,culture medium is:MS+BA0.3mg/L+NAA1.5mg/L+25g/L sucrose+7g/L agar (pH5.8); The most suitable medium for callus proliferation and induce differentiation to adventitious bud is:MS+2,4-D1.5mg/L+BA0.1mg/L+30g/L sucrose+8g/L agar (pH5.8); The optimal rooting medium is:1/2MS+0.5mg/L IBA+1mg/LPP333+0.3mg/LBA+8g/Lagar (PH5.8)。... |
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