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

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: , , , , , , , , , , , , , , . 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, , , , together with , were the most unstable genes in both the turn over and withering treatments. , , , and were the top four reference genes in the turn over treatment, while , , , , and 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.