Screening and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR under PEG, NaCl and ZnSO Treatments in .

Real-time quantitative PCR (RT-qPCR) has a high sensitivity and strong specificity, and is widely used in the analysis of gene expression. Selecting appropriate internal reference genes is the key to accurately analyzing the expression changes of target genes by RT-qPCR. To find out the most suitable internal reference genes for studying the gene expression in under abiotic stresses (including drought, salt, and ZnSO treatments), seven different tissues of , as well as the roots, stems, and leaves of under the abiotic stresses were used as test materials, and 15 candidate internal reference genes were screened based on the transcriptome data via RT-qPCR. Then, the expression stability of the candidate genes was comprehensively evaluated through the software geNorm (v3.5), NormFinder (v0.953), BestKeeper (v1.0), and RefFinder. The best internal reference genes and their combinations were screened out according to the analysis results. and were the best reference genes under drought stress. Under salt stress, , , , and were the most stable reference genes. Under heavy metal stress, and were the most suitable reference genes. and were the most suitable internal reference genes in the different tissues of . In addition, , , , and were the most suitable internal reference genes for the abiotic stresses and the different tissues of . The expression patterns of and were analyzed by using the selected stable and unstable reference genes. This further verified the reliability of the screened internal reference genes. This study lays the foundation for the functional analysis and regulatory mechanism research of genes in .