Screening And Evaluation Of Reference Genes In The QRT-PCR Analysis Schisandra Chinensis

Real-time quantitative fluorescence qRT-PCR is the most widely used method for gene expression analysis.It has many advantages,such as simple and rapid operation,sensitivity and strong specificity.However,when qRT-PCR is used for gene expression measurement,it is very necessary to screen and verify internal reference genes.So far,there are few studies on gene expression analysis of S.chinensis,and no studies on internal reference genes of S.chinensis.In this study,S.chinensis with different genotypes,different tissues and different abiotic stress treatments were used as test materials to extract RNA and reverse transcribed into c DNA.Ten candidate reference genes,including GPN1,PP2A15,RPL6,RPL15,RPL21,UBC2,UBC11 and UBQ12,were selected by referring to the traditional reference genes in plants.Primers were designed for amplification by qRT-PCR.Statistical software ge Norm,Norm Finder and Best Keeper and △ Ct algorithm were used for data analysis to evaluate the expression stability of each candidate reference gene,so as to select the most suitable reference gene.The main results obtained are:1.RPL6 was the best choice of internal reference gene in S.chinensis of different varieties and tissues.2.In abiotic stress treatment,UBQ12 can be used as the best reference gene in S.chinensis treated with salt and alkali stress.RPL6 was the best choice of reference gene in S.chinensis treated with drought stress.3.In the three abiotic stress sample sets of S.chinensis,UBC2 showed excellent performance and could be used as the best reference gene.In all samples of S.chinensis,UBQ12 showed excellent expression stability and was the best choice for reference genes.Combined with the results of this experiment,it can be seen that the expression of candidate reference genes is different in different sample sets.Under different experimental conditions,there is no reference gene expression can be maintained constant.Therefore,in the actual test operation,the selection of internal reference genes should be carried out according to the specific test materials and test conditions to ensure the accuracy of the test data.In summary,this study used real-time fluorescence quantitative technology to screen the optimal internal reference genes for Schisandra under different experimental conditions,which laid a foundation for further research on molecular biology of S.chinensis in the future.