刨花润楠SRAP-PCR体系建立与优化  

Establishment and Optimization of SRAP-PCR System for Machilus pauhoi

作　　者：周鹏[1,2] 林玮[2] 周祥斌[2] 陈晓阳[2] 

机构地区：[1]广东生态工程职业学院,广东广州510520 [2]华南农业大学林学与风景园林学院/广东省森林植物种质创新与利用重点实验室,广东广州510642

出　　处：《林业与环境科学》2017年第4期29-33,共5页Forestry and Environmental Science

基　　金："十二五"国家科技支撑项目"刨花润楠和黄樟良种选育研究"(2012BAD01B04);广东省林业科技创新项目"楝科;樟科优质速生树种良种选育和高效栽培技术研究与示范"(2011KJCX002)

摘　　要：以刨花润楠(Machilus pauhoi)1.5 a生小苗幼嫩叶片为试材,对影响刨花润楠SRAP-PCR扩增的模板DNA量、引物、dNTP和Mg^(2+)体积摩尔浓度、Taq DNA聚合酶、退火温度6个主要因素进行优化。结果表明,SRAP-PCR的最佳反应体系为:25μL的SRAP-PCR反应体系中,2.5μL 10×PCR buffer、模板DNA量60 ng、Mg^(2+)2.0 mmol/L、d NTP 0.225 mmol/L、引物0.3μmol/L和Taq DNA聚合酶1.25 U。对优化的反应体系和扩增程序的验证结果表明,优化的刨花润楠SRAP-PCR反应体系和扩增程序是稳定可行的。Machilus pauhoi is a tree specie with variety of economic value and development prospects. This study aimed to establish an optimized SRAP-PCR system for M. pauhoi, and the young leaves of the 1.5 years old seedlings were used as test materials. Six quality factors including the template DNA, primer concentration, d NTP concentration, Mg^2＋concentration, Taq DNA polymerase, and annealing temperature were optimized for M. pauhoi SRAP-PCR assay. The obtained results suggested a optimized reaction system of SRAP-PCR（total 25 μL） involving 2.5 μL 10×PCR buffer, 60 ng DNA, 2.0 mmol/L Mg^2＋, 0.225 mmol/L d NTP, 0.3 μmol/L primer, 1.25 U Taq DNA polymerase. The verification results showed that the optimized SRAP-PCR reaction system and amplification program were stable and feasible.

关 键 词：刨花润楠 SRAP-PCR 体系优化 

分 类 号：S718.43[农业科学—林学] S792.23