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For quantitative real-time PCR (qRT-PCR) analysis, the key prerequisite that determines result accuracy is the selection of appropriate reference gene(s). Goji ( Lycium barbarum  L.) is a multi-branched shrub belonging to the Solanaceae family. To date, no systematic screening or evaluation of reference gene(s) in Goji has been performed. In this work, we identified 18 candidate reference genes from the transcriptomic sequencing data of 14 samples of Goji at different developmental stages and under drought stress condition. The expression stability of these candidate genes was rigorously analyzed using qRT-PCR and four different statistical algorithms: geNorm, BestKeeper, NormFinder and RefFinder. Two novel reference genes  LbCML38  and  LbRH52  showed the most stable expression, whereas the traditionally used reference genes such as  LbGAPDH, LbHSP90  and  LbTUB  showed unstable expression in the tested samples. Expression of a target gene  LbMYB1  was also tested and compared using optimal reference genes  LbCML38  and  LbRH52 , mediocre reference gene  LbActin7 , and poor reference gene  LbHSP90  as normalization standards, respectively. As expected, calculation of the target gene expression by normalization against  LbCML38, LbActin7  or  LbHSP90  showed significant differences. Our findings suggest that  LbCML38  and  LbRH52  can be used as reference genes for gene expression analysis in Goji.

LbCML38 and LbRH52, two reference genes derived from RNA-Seq data suitable for assessing gene expression in Lycium barbarum L.