红树植物海漆ISSR条件的优化  

Optimization of ISSR Analysis for Excoecaria agollocha L.

作　　者：张志红[1] 谈凤笑[1] 何航航[1] 吴令辉[1] 钟才荣 施苏华[1] 

机构地区：[1]中山大学生命科学学院,广东广州510275 [2]东寨港国家级自然保护区,海南海口571129

出　　处：《中山大学学报（自然科学版）》2004年第2期63-66,共4页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家自然科学基金资助项目(30070053;30230030);广东省自然科学基金资助项目(001223);教育部博士点基金资助项目(20010558013)

摘　　要：在利用ISSR分析对海漆Excoecariaagallocha的遗传多样性进行了研究。为获得清晰、重复性的ISSR扩增结果,对影响ISSR＿PCR的多个因素,包括DNA的提取、模板浓度、TAQ酶的选择与用量、Mg2+和dNTP浓度等进行了比较、优化,确定了海漆ISSR＿PCR分析最适宜的扩增条件:10μLPCR反应体积中,1×Taq酶配套缓冲液(10mmol LTris·HCl,pH9 0,50mmol LKCl,0 1%TritonX-100),0 6～0 7UTaqDNA聚合酶(上海申能博彩生物科技有限公司),0 2μmol L引物,0 2mmol LdNTP,1 5～2 0mmol LMgCl2,10ng模板DNA。Excoecaria agallocha L.is a medical mangrove plant of Euphorbiaceae. The factors which affect the ISSR analysis in the study of the genetic diversity of\$ E\^agallocha\$, such as genomic DNA extraction, concentrations of template DNA, Taq DNA polymerase, Mg^(2+) and dNTP, etc were examined. The optimal ISSR_PCR conditions in our experiments were as the following: 1×Taq buffer (10 mmol/L Tris·HCl, pH 9.0, 50 mmol/L KCl, 0.1% Triton X_100), 10 ng template DNA, 0.6～0.7 U Taq DNA(produced by Shenneny Bocai Company), 0.2 μmol/L primer, 1.5～2.0 mmol/L MgCl2, 0.2mmol/L dNTP. The length of amplification gave clear and reproducible banding patterns. The length of bands ranged from 200 to 2 500 bp and exhibited a high level of polymorphism.

关 键 词：海漆Excoecaria agallocha ISSR 优化 红树植物 

分 类 号：Q503[生物学—生物化学] Q946-33