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

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 ap...

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Autores principales: Zhou, Zi-wei, Deng, Hui-li, Wu, Qing-yang, Liu, Bin-bin, Yue, Chuan, Deng, Ting-ting, Lai, Zhong-xiong, Sun, Yun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6360078/
https://www.ncbi.nlm.nih.gov/pubmed/30723635
http://dx.doi.org/10.7717/peerj.6385
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author Zhou, Zi-wei
Deng, Hui-li
Wu, Qing-yang
Liu, Bin-bin
Yue, Chuan
Deng, Ting-ting
Lai, Zhong-xiong
Sun, Yun
author_facet Zhou, Zi-wei
Deng, Hui-li
Wu, Qing-yang
Liu, Bin-bin
Yue, Chuan
Deng, Ting-ting
Lai, Zhong-xiong
Sun, Yun
author_sort Zhou, Zi-wei
collection PubMed
description 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.
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spelling pubmed-63600782019-02-05 Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis) Zhou, Zi-wei Deng, Hui-li Wu, Qing-yang Liu, Bin-bin Yue, Chuan Deng, Ting-ting Lai, Zhong-xiong Sun, Yun PeerJ Agricultural Science 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. PeerJ Inc. 2019-01-31 /pmc/articles/PMC6360078/ /pubmed/30723635 http://dx.doi.org/10.7717/peerj.6385 Text en © 2019 Zhou et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , 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) and either DOI or URL of the article must be cited.
spellingShingle Agricultural Science
Zhou, Zi-wei
Deng, Hui-li
Wu, Qing-yang
Liu, Bin-bin
Yue, Chuan
Deng, Ting-ting
Lai, Zhong-xiong
Sun, Yun
Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
title Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
title_full Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
title_fullStr Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
title_full_unstemmed Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
title_short Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)
title_sort validation of reference genes for gene expression studies in post-harvest leaves of tea plant (camellia sinensis)
topic Agricultural Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6360078/
https://www.ncbi.nlm.nih.gov/pubmed/30723635
http://dx.doi.org/10.7717/peerj.6385
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