花生SRAP反应体系的建立与优化

Establishment and Optimization of SRAP Reaction Conditions for Arachis hypogaea L.

[目的]为研究花生居群遗传多样性奠定基础。[方法]以汕油162和sunolicn 95R为模板对花生进行SRAP扩增,研究模板DNA浓度、Mg2+浓度、引物浓度和TaqDNA聚合酶的用量对SRAP扩增效果的影响,探索花生SRAP反应的最佳条件。[结果]在一定范围内,模板DNA含量和TaqDNA聚合酶的用量对花生SRAP扩增结果的影响较小。当Mg2+浓度为2.5 mmol/L时,扩增效果最佳。当Mg2+浓度逐渐降低时,扩增条带逐渐变弱。引物浓度为0.5 mmol/L时能够扩增出清晰、重复性好的条带。花生SRAP反应的最佳条件为:模板DNA含量为100 ng,Mg2+浓度为2.5 mmol/L,引物浓度为0.5 mmol/L,TaqDNA聚合酶的用量为1 U。[结论]与AFL P技术相比,该研究中所建立的花生SRAP反应体系更为简便和可靠。

[ Objective] The aim of the research was to lay the foundation for studying the genetic diversity of Arachis hypogaea L. populations. [ Method] With Shanyou 162, stmoliec 95R as template, SRAP amplification was made on A. hypogaea to study the effects of the concn, of template DNA, Mg^2 ＋ concn. ,primer conch, and the dosage of Taq DNA polymerase on the effect of SRAP amplification. And the optimum conditions for SRAP reaction of A. hypogaea were discussed. [ Result ] In a certain range, template DNA content and the dosage of Taq DNA polymerase had little effects on the results of SRAP amplification for A. hypogaea. When Mg^2 ＋ was 2.5 mmol/L, the amplification results were best. When Mg^2 ＋ concn, was gradually decreased, the amplified bands were gradually weakened. When the primer was 0.5 mmol/L, the clear bands with good repeatability could be amplified. The optimum conditions for SRAP reaction of A. hypogaea were as follows： template DNA content was 100 ng, Mg^2＋ was 2.5 mmol/L, primer was 0.5 mmol/L and the dosage of Taq DNA polymerase was 1 U. [ Conclusion] Compared with AFL P technology, the SRAP reaction system for A. hypogaea established in this research was more convenient and reliable.