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Reverse transcription quantitative PCR (RT-qPCR) is a technique widely used to investigate the expression of genes. An appropriate reference gene (RG) is essential for RT-qPCR analysis to obtain accurate and reliable results.  Caragana intermedia  plays an important role in afforestation as a bush. However, due to the lack of appropriate RGs, the research on development-related genes is limited. In this study, the selection for suitable RGs of different organs at various development stages to normalize the results of RT-qPCR about development-related genes was performed. To test the expression stability across all samples, we used the software algorithms such as geNorm, NormFinder, BestKeeper, and RefFinder to evaluate all the candidate RGs. Our results showed that  CiEF1α  was the most stable RG with little fluctuation among all samples. In addition,  CiGAPDH  in roots,  CiSKIP1  in stems and leaves, and  CiEF1α  in different organs were selected as the most stable RGs. To confirm the applicability of the most stable RGs, the relative expression of  CiWRKY17  was normalized using different candidate RGs. Taken together, our research laid a foundation for the study of development-related genes in  C. intermedia .

Selection and validation of reference genes for RT-qPCR analysis of different organs at various development stages in Caragana intermedia