Identification and validation of reference genes for RT-qPCR analysis in under Cd stress.

is a widely used ornamental garden and important medicinal plant. Our previous studies have shown that it exhibits significant uptake and translocation capacity under Cd stress compared to other species. Gene expression is studied using RT-qPCR; however, there are no reference genes have been found for under Cd stress. In this investigation, thirteen possible reference genes from previous studies and our transcriptome were screened using RT-qPCR in the leaves and roots of Cd-stressed plants. The findings revealed that and were the best reference genes for roots with and without Cd stress, whereas and were the best reference genes for leaves. Among the different tissues without Cd stress, and exhibited the best results, whereas and exhibited the best results under Cd stress. The most stable reference genes in the leaves and roots were and , respectively, under and without Cd stress, and was the most unstable. Finally, three metal ion response genes, , and were detected using RT-qPCR and compared with the transcriptome data to further confirm the reliability of the chosen genes. This study identified suitable reference genes for under Cd-stress conditions.