白芨SCoT分子标记技术体系的建立及其在快繁苗遗传稳定分析中的应用

Establishment and Application of SCoT Molecular Marker System for Genetic Stability of Rapid Propagation Plants of Bletilla striata

以4个不同继代的白芨快繁苗为试材,采用L_(16)(4~5)正交实验设计,建立了白芨SCoT-PCR的反应体系,并利用该体系分析不同继代的白芨快繁苗遗传稳定性。结果表明:白芨的SCoT-PCR最佳反应体系(20μL)中包含2.500 mmol·L^(-1) Mg^(2+),0.25mmol·L^(-1) dNTPs,1.50 U Taq DNA聚合酶,1.000μmol·L^(-1)引物,10ng模板DNA。遗传稳定性分析发现,SCoT引物均能在4个不同继代的白芨快繁苗中扩增出清晰、丰富而稳定的条带,但每个引物的条带均无多态性。说明白芨快繁苗并未发生变异,其遗传稳定性较好。

SCoT-PCR reaction system established by L16（4^5）orthogonal experimental design was targeted to sub-culture rapid propagation of Bletilla striata.Genetic stabilities of those sub-culture propagated plantlets were evaluated by this system.The results showed that an optimal reaction system of Bletilla striata contained 2.500 mmol·L^-1 Mg^2＋,0.25 mmol·L^-1 dNTPs,1.50 U Taq DNA polymerase,1.000μmol·L-（-1） primer as well as DNA template 10 ng.Also,clear,rich and stable bands were observed in each sub-culture rapid propagation based on SCoT-PCR amplification system without polymorphism.All in all,rapid propagation of Bletilla striata kept genetic stability without mutation from generation to generation.