伯乐树ISSR分子标记体系的建立  

Establishment of ISSR-PCR Reaction System for Bretschneidera sinensis

作　　者：谢正成 楼雄珍[2] 姚理武 黄华宏[2] 林二培[2] 骆文坚 陈春梅 

机构地区：[1]浙江省庆元县林业局,浙江庆元323800 [2]浙江农林大学,浙江临安311300 [3]浙江省林业种苗管理总站,浙江杭州310020

出　　处：《浙江林业科技》2011年第1期11-15,共5页Journal of Zhejiang Forestry Science and Technology

基　　金：浙江省重大科技专项"浙江省珍稀濒危林木种质资源收集保存与利用关键技术研究及基因库建设"(2006C12059-4);浙江省科技重点项目"浙江省珍稀濒危树种现代保育关键技术研究"(2006C22064)

摘　　要：从浙江省庆元县等地采集23份野生伯乐树单株叶片为试验材料,采用单因素对比试验和正交优化试验研究了模板DNA、Taq DNA聚合酶的用量,以及Mg^(2+)、dNTP、引物浓度和退火温度对PCR扩增的影响,建立伯乐树适宜的ISSR-PCR扩增体系,即20μL体系中含90ng模板DNA,1×PCR buffer,1.0mmol/L Mg^(2+),0.15mmol/LdNTP,0.5μmol/L引物,0.75U Taq DNA聚合酶,退火温度为50℃。The aim of this study was to establish and optimize the amplification system of ISSR-PCR for Bretschneidera sinensis. 23 samples of B. sinensis leaf were collected from Qingyuan of Zhejiang province for the test. The influencing factors of ISSR-PCR, such as DNA template, Taq DNA polymerase, Mg2＋, dNTPs, primer and annealing temperature were optimized by test of single factor and orthogonal design. The results indicated that suitable ISSR-PCR system for B. sinensis was established, namely 20μL reaction system containing 90 ng template DNA, 1 x PCR buffer, 1.0 mmol/L Mg2＋, 0.15mmol/L dNTPs, 0.5 μmol/L primers, 0.75U Taq DNA polymerase. The optimal annealing temperature was 50℃ .

关 键 词：伯乐树 ISSR 分子标记 

分 类 号：S718.4[农业科学—林学]