花生SRAP反应体系的建立与优化

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。

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

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,sunoliec 95R as template,SRAP amplification was made on A.hypogaea to study the effects of the concn.of template DNA,Mg2 concn.,primer concn.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 Mg2 was 2.5 mmol/L,the amplification results were best.When Mg2 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,Mg2 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.