荧光定量PCR技术优化及在草莓上的应用

Application of optimized fluorescence quantitative PCR technology for the research of strawberry

【目的】为优化荧光定量PCR技术体系并在草莓研究中应用。【方法】以二倍体草莓(Fragaria vesca‘Ruegen’)为试材,草莓当中两个β-肌动蛋白基因家族成员Actin1和Actin2为内参基因,对比分析10μL和20μL反应体系条件下荧光定量PCR扩增反应。【结果】内参基因引物组合Actin1的扩增效率等指标优于Actin2;10μL反应体系中的扩增效率等指标优于20μL反应体系;内参基因Actin1在10μL反应体系下是草莓最优的荧光定量PCR技术体系。【结论】优化了草莓荧光定量PCR体系并应用该技术体系检测了草莓CrRLK1Ls家族成员的时空表达情况。

【Objective】In order to establish and optimize quantitative real-time PCR system for studying gene expression of strawberry.【Methods】Twoβ-actin genes（Actin1and Actin2）in strawberry（Fragaria vesca ‘Ruegen＇）were used reference genes in amplification reaction.The amplification efficiency and standard curve were studied in 10μL and 20μL reaction system conditions.【Results】The results showed that the amplification efficiency and standard curve related index of Actin1 were better than that of Actin2.The amplification efficiency and standard curve related index in 10μL reaction system were better than that of 20μL reaction system.It is optimal quantitative real-time PCR reaction system for strawberry that Actin1 as reference gene in 10μL reaction system.The optimal quantitative real-time PCR reaction system was used to determine the spatiotemporal expression of CrRLK1 Ls subfamily in strawberry.【Conclusion】The fluorescence quantitative PCR technology was optimized and used to studied spatiotemporal expression of Strawberry.The CrRLK1 Ls subfamily may play the significant role in regulating strawberry fruit development.