46份菌草种质资源PEPC基因的PCR-RFLP多样性分析

Polymorphism Analysis of PEPC Gene Family among 46 Juncao Germplasms by PCR-RFLP

根据近缘种间同源基因序列保守的原理,设计20对可用于扩增菌草PEPC基因的特异性PCR反应引物。利用PCR-RFLP技术对46份菌草种质资源进行PEPC基因多样性分析。结果表明：12对引物可对供试材料实现有效特异性扩增,PCR产物经限制性核酸内切酶Hae III酶切,在46份材料上共给出176（平均每对引物14.7）个变异类型。UPGMA聚类分析结果表明,材料间遗传相似系数（SM）在0.86-1,在遗传相似系数为0.865时,46份材料可分为四大类群,这表明PCR-RFLP技术可有效用于菌草资源的遗传多样性分析。

A tot al of 20 pa irs of specific primers were designed to amplify the phosphoenolpyruvate carboxylase（PEPC） gene family of JUNCAO germplasms. PCR-Restriction fragment length polymorphism（PCR-RFLP） technique was applied to study the polymorphism of PEPC gene family among 46 JUNCAO germplasms. The results showed that 12 of 20 primer pairs gave good amplification patterns, and digestion of these PCR products by Hae III gave a total of 176（14.7 per primer pair in average） different patterns among 46 JUNCAO germplasms. The genetic similarity coefficient（SM） among all accessions was ranged from 0.86 to 1. The 46 JUNCAO germplasms were clustered into 4 groups by UPGMA when genetic similarity coefficient was 0.865. The study indicated that PCR-RFLP technique could be efficiently used for analysis of genetic diversity of JUNCAO germplasms.