Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
- Published
- Accepted
- Subject Areas
- Agricultural Science, Biotechnology, Molecular Biology
- Keywords
- reference genes, RT-qPCR, post-harvest leaves, Camellia sinensis, CsLOX1
- Copyright
- © 2018 Zhou et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
- Cite this article
- 2018. Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis) PeerJ Preprints 6:e27287v1 https://doi.org/10.7287/peerj.preprints.27287v1
Abstract
Tea is one of three major non-alcoholic beverages that are popular all around the world. The economic value of the final tea product largely depends on the post-harvest physiology of tea leaves. The utilization of quantitative reverse transcription polymerase chain reaction (RT-qPCR) 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 have 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 witheringtreatments. 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 four best reference genes in the witheringgroup. The expression level of lipoxygenase (LOX) 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.
Author Comment
This is a submission to PeerJ for review.