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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (2): 20-24.

所属专题： 食用菌； 食用菌

• 生物技术科学 • 上一篇    下一篇

香菇SSR-PCR技术体系的建立及其在遗传多样性分析中的初步应用

肖扬   

  • 收稿日期:2008-10-17 修回日期:2008-11-27 出版日期:2009-01-20 发布日期:2009-01-20

Establishment of SSR-PCR Technological System and Its Preliminary Application in Genetic Diversity Analysis for Lentinula edodes

XIAO Yang   

  • Received:2008-10-17 Revised:2008-11-27 Online:2009-01-20 Published:2009-01-20

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摘要： 摘 要: 利用ISSR-抑制PCR法结合Primer 3软件开发香菇特异性的SSR引物,并采用L16（45）正交试验设计,对影响香菇SSR-PCR的主要因素进行了优化筛选，建立了最佳的SSR-PCR反应体系。在20μl反应体系中，包含1.5mmol/L Mg2+，75ng模板DNA，0.25mmol/L dNTPs，0.50μmol/L引物及1.5U Taq酶。梯度PCR试验筛选得到相应引物的最佳退火温度为62℃。以该体系为基础，应用5对引物扩增8个香菇菌株的基因组DNA，均能获得理想结果，聚类分析结果能较好地反映供试菌株的地理来源关系。以0.5 的相似性为分割点，8个菌株可分成2大类群。类群Ⅰ主要由北方菌株组成，类群Ⅱ由南方菌株组成。

关键词: 早籼稻, 早籼稻, 塑盘育秧抛栽, 群体发育指标, 调控技术

Abstract: Abstract: SSR primers of Lentinula edodes were developed using ISSR-suppression PCR method and Primer 3 program, and L16（45）orthogonal design was used to optimize main factors affecting SSR-PCR amplification system. The optimal SSR-PCR system was established as follows, within 20μl reaction system, there were 1.5 mmol/L Mg2+, 75 ng template DNA, 0.25 mmol/L dNTPs, 0.50 μmol/L primer and 1.5U Taq DNA polymerase. The optimal annealing temperature was determined as 62℃ by gradient PCR. Based on the optimal system, suitable results were obtained when five primer pairs were used to amplify genomic DNA of eight strains .Cluster analysis well reflected geographical original relation of these strains. The eight strains were classified into two groups using 0.5 similarity as a cut-off point. GroupⅠmainly consisted of strains from North China, while group Ⅱ comprised those from South China.