Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
| Autor: | Zi-wei Zhou, Yun Sun, Ting-ting Deng, Zhongxiong Lai, Hui-li Deng, Qing-yang Wu, Chuan Yue, Bin-bin Liu |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
lcsh:Medicine Plant Science Biology 01 natural sciences Camellia sinensis General Biochemistry Genetics and Molecular Biology CsLOX1 03 medical and health sciences Transcription (biology) Reference genes Gene expression Post-harvest leaves Agricultural Science Molecular Biology Gene 030304 developmental biology 0303 health sciences Manufacturing process business.industry General Neuroscience RT-qPCR lcsh:R Plant physiology food and beverages General Medicine Biotechnology Reverse transcription polymerase chain reaction General Agricultural and Biological Sciences business 010606 plant biology & botany |
| Zdroj: | PeerJ, Vol 7, p e6385 (2019) PeerJ |
| DOI: | 10.7287/peerj.preprints.27287v1 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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