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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (13): 241-245.

所属专题： 园艺

• 农学 农业基础科学 • 上一篇    下一篇

橡胶树SAM-SSR体系的优化

于 飞,,冯素萍,,童和林,,武耀廷   

  • 收稿日期:2009-04-16 修回日期:2009-05-14 出版日期:2009-07-05 发布日期:2009-07-05

Optimization of SAM-SSR System in Rubber Tree

Yu Fei,, Feng Suping,, Tong Helin,, Wu Yaoting   

  • Received:2009-04-16 Revised:2009-05-14 Online:2009-07-05 Published:2009-07-05

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摘要： 以橡胶树品种热研88-13为试材,研究了橡胶树SAM-SSR法中各个过程（酶切、连接、抑制性和预扩增）中各因素对SAM法结果的影响。实验结果表明：5U MseⅠ和5U PstⅠ双酶切1h为最合适酶切浓度、方式和时间; 100~300ng为适宜的DNA模板浓度； 合适的连接温度和时间为16℃连接过夜；连接产物和抑制性PCR产物分别稀释20倍，为抑制性PCR和预扩增PCR的适宜模板浓度。 SAM扩增产物用8%的非变性聚丙烯酰胺凝胶检测，结果显示，扩增条带分布在 200~750bp之间，能够满足建立橡胶树SSR基因组文库的需要。

关键词: 棉花, 棉花, 红腐病菌, 酶活性, 抗病性

Abstract: Reyan88-13, a superior clone of rubber tree, was used as material to study the affects of SAM-SSR (digestion, linkage, suppressed amplification and pre-amplification PCR). The results showed that the optimal digest concentration, method and time of enzyme were 5U MseⅠand 5U PstⅠ,double digest for 1h; the optimal concentration of DNA template was 100~300 ng/ul; the optimal temperature and time of linkage were 16℃ overnight; And the suppressed amplification and pre-amplification PCR were set up based the productions of linkage and suppressed amplification diluted by 20 times respectively. The productions of SAM amplification were detected by 8% Polyacrylamide Gel Electrophoresis and the results showed, the amplified productions ranged from 200bp to 750bp and could be used to construct the SSR genomic library for rubber tree.