油茶SRAP-PCR反应体系的优化.pdfVIP

林业科学研究 2010 ， 23(2) :302 307 Forest Research 文章编号:1∞1-1498 (2010 )02 -0302-06 油茶 SRAP-PCR 反应体系的优化 郑婷婷1 气林萍2 ， 王开良2 ，姚小华2 ， 杨水平1* (1.商南大学资源环境学院，重庆 4阳716; 2. 中国林业科学研究院亚热带林业研究所，浙江富阳 3114∞) 关键词:油茶;5RAP; 正交设计;体系优化 中图分类号:5794.4 文献标识码:A Optimization of SRAP-PCR System for Camellia oleifera 2 ZHENG Ting-tingl ,2 , UN Pini , WANG Kai-liang2 , YAO Xiao峭hua ， YANG Shui-pingl (1. College of Resources and Environmenl of Southw耐 China University , Chon阴ing 4∞716 ， China; 2 , Research lnstilute of Subtropical Fores町， Chinese Academy of For田町， Fuyang 3114∞， Zhejian毡， China) Abstract: Camellia oleifera is one of the important oil tree species in south China , and C. oleifera indus盯 is quickly developed with the support of the national policies in 陀cent years. The disorder of ι oleifera varieties is one of the key issues restricting the development of C. ole拚ra industry. Because of high polymorphism , good repeatability , less use of DNA and so on , SRAP as a new marker was used in identification of cultivaI毡， analysis of genetic resources and genetic diversity in recent ye创8. In this paper , the orthogonal design was used to optimize 2 SRAP-PCR system for C. oleifera by 5 factors (Mg + , dNTPs , primer , Taq polymerase , DNA template) 四d4 levels , respectively. The data were analyzed by so仇W盯e SPSS V13. O. A suitable SRAP-PCR system (20 J.LL) was established as: 75 ng DNA template , 1. 5 mmol • L -1 Mg2 + , 0.15 mmol • L -1 dNTPs , 1U Taq DNA pol严ner幽e ， 0.4μmol • L -1 primer , 1 x PCR buffer. The result of optimal SRAP-PCR system wiII provide