太子参RAPD反应体系的优化  

Optimization of the RAPD reaction system for Radix pseudostellariae

作　　者：史辉 董晓菲 叶祖云 SHI Hui;DONG Xiao-fei;YE Zu-yun(College of Life Science,Ningde Normal University,Ningde,Fujian 352100,China;Fujian Higher Education Research Center for Local Biological Resources in Ningde City,Ningde,Fujian 352100,China)

机构地区：[1]宁德师范学院生命科学学院,福建宁德352100 [2]闽东特色生物资源福建省高校工程研究中心,福建宁德352100

出　　处：《宁德师范学院学报（自然科学版）》2022年第1期58-63,共6页Journal of Ningde Normal University(Natural Science)

基　　金：福建省自然科学基金项目(2016J01169);宁德师范学院科研发展资金项目(2020FZ05)。

摘　　要：为探讨太子参 RAPD-PCR 的最佳反应体系条件,通过单因子试验对影响太子参 RAPD-PCR 扩增效果的因素,如模板 DNA 浓度、Taq DNA 聚合酶浓度、d NTPs浓度、引物浓度、延伸时间和循环数进行优化 .确立可用于太子参 RAPD-PCR 分析最适宜的反应体系,即 20 μL 反应体系中含 28 ng 模板 DNA,0.7 U Taq 酶,0.8μmol·L;引物,0.22 mmol·L;d NTPs;PCR 扩增延伸时间为 100 s,循环数 40 次 . 稳定性试验表明,该体系能在不同太子参品种间扩增出清晰明亮、稳定性、多态性好的条带.优化后的体系可用于太子参种质资源的鉴定.The optimum condition of RAPD-PCR for Radix pseudostellariae was studied. The factors which af-fected the RAPD-PCR amplification on Radix pseudostellariae such as template DNA concentration, Taq DNA polymerase, dNTPs, primers, extension time and cycle number were optimized through a combination of single factor experiments. The results showed that the optimal reaction system for RAPD-PCR of Radix pseudostellariae was performed in a 20 μL reaction volume with 28 ng template DNA, 0.7 U Taq DNApolymerase, 0.8μM primers, 0.22 mmol·L-1dNTPs, 100 s extended time for PCR, and 40 PCR cycles. Stability test results showed that it produced clear and bright bands with good stability, and the polymorphism can be used to distinguish dif-ferent varieties of Radix pseudostellariae. The optimized system can be used for the identification of germplasm resources for Radix pseudostellariae.

关 键 词：太子参 RAPD 反应体系 单因子试验 

分 类 号：S567[农业科学—中草药栽培]