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Tea is one of three major non-alcoholic beverages that are popular all around the world. The economic value of tea product largely depends on the post-harvest physiology of tea leaves. The utilization of quantitative reverse transcription polymerase chain reaction is a widely accepted and precise approach to determine the target gene expression of tea plants, and the reliability of results hinges on the selection of suitable reference genes. A few reliable reference genes have been documented using various treatments and different tissues of tea plants, but none has been done on post-harvest leaves during the tea manufacturing process. The present study selected and analyzed 15 candidate reference genes:  Cs18SrRNA ,  CsGADPH ,  CsACT ,  CsEF-1α ,  CsUbi ,  CsTUA ,  Cs26SrRNA ,  CsRuBP ,  CsCYP ,  CselF-4α ,  CsMON1 ,  CsPCS1 ,  CsSAND ,  CsPPA2 ,  CsTBP . This study made an assessment on the expression stability under two kinds of post-harvest treatment, turn over and withering, using three algorithms—GeNorm, Normfinder, and Bestkeeper. The results indicated that the three commonly used reference genes,  CsTUA ,  Cs18SrRNA ,  CsRuBP , together with  Cs26SrRNA , were the most unstable genes in both the turn over and withering treatments.  CsACT ,  CsEF-1α ,  CsPPA2 , and  CsTBP  were the top four reference genes in the turn over treatment, while  CsTBP ,  CsPCS1 ,  CsPPA2 ,  CselF-4α , and  CsACT  were the five best reference genes in the withering group. The expression level of lipoxygenase genes, which were involved in a number of diverse aspects of plant physiology, including wounding, was evaluated to validate the findings. To conclude, we found a basis for the selection of reference genes for accurate transcription normalization in post-harvest leaves of tea plants.

Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)