披碱草属物种AFLP体系优化

试验对内切酶EcoRⅠ和MseⅠ的用量和酶切时间、预扩和选扩过程中模板稀释倍数以及扩增引物浓度分别进行了优化,并采用了Bassam法和Sanguinetti银染法进行了比较分析。结果表明:采用3UEcoRⅠ和MseⅠ这两种酶在37℃下酶切3 h,预扩模板稀释10倍和1.5 ng/μL引物进行预扩,选扩模板稀释20倍和2 ng/μL引物进行选扩,采用Bassam法进行银染,能够得到较好的试验结果,为AFLP进一步在披碱草属中的应用提供了参考。

Optimization of AFLP in Elymus (Poaceae)

The key factors affecting the AFLP analysis in Elymus were optimized,including the digesting time and the content of the EcoRⅠ and MseⅠ,diluted ratio and concentrations of primers in the pre-amplification and selective amplification.Furthermore,the methods of Bassam and Sanguinetti silver staining were also compared in the staining quality.The optimal reaction system of AFLP were established,which including digest with 3 units of each enzymes at 37 ℃ for 3 h,pre-amplification performed with templates diluted 10 times and 1.5 ng/μL primers,selective amplification performed with templates diluted 20 times and 2 ng/μL primers,as well as stained with Bassam silver staining.The results could apply for the further study on AFLP in Elymus.