油茶SRAP-PCR反应体系的优化

Camellia oleifera is one of the important oil tree species in south China,and C.oleifera industry is quickly developed with the support of the national policies in recent years.The disorder of C.oleifera varieties is one of the key issues restricting the development of C.oleifera industry.Because of high polymorphism,good repeatability,less use of DNA and so on,SRAP as a new marker was used in identification of cultivars,analysis of genetic resources and genetic diversity in recent years.In this paper,the orthogonal design was used to optimize SRAP-PCR system for C.oleifera by 5 factors(Mg2+,dNTPs,primer,Taq polymerase,DNA template) and 4 levels,respectively.The data were analyzed by software SPSS V13.0.A suitable SRAP-PCR system(20 μL) was established as: 75 ng DNA template,1.5 mmol·L-1 Mg2+,0.15 mmol·L-1 dNTPs,1U Taq DNA polymerase,0.4 μmol·L-1 primer,1×PCR buffer.The result of optimal SRAP-PCR system will provide a foundation for the identification of C.oleifera cultivars.