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Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus

The role of Melatonin in influencing diverse genes in plants has gained momentum in recent years and many reports have employed qRT-PCR for their quantification. Relative quantification of gene expression relies on accurate normalization of qRT-PCR data against a stably-expressing internal reference...

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Autores principales: Sheshadri, S. A., Nishanth, M. J., Yamine, V., Simon, Bindu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5799177/
https://www.ncbi.nlm.nih.gov/pubmed/29403001
http://dx.doi.org/10.1038/s41598-018-20474-2
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author Sheshadri, S. A.
Nishanth, M. J.
Yamine, V.
Simon, Bindu
author_facet Sheshadri, S. A.
Nishanth, M. J.
Yamine, V.
Simon, Bindu
author_sort Sheshadri, S. A.
collection PubMed
description The role of Melatonin in influencing diverse genes in plants has gained momentum in recent years and many reports have employed qRT-PCR for their quantification. Relative quantification of gene expression relies on accurate normalization of qRT-PCR data against a stably-expressing internal reference-gene. Although researchers have been using commonly available reference-genes to assess Melatonin-induced gene expression, but to-date, there have been no attempts to validate the reference-gene stability under Melatonin-supplementation in planta. In this study, we performed stability assessment of common reference-genes under Melatonin-supplementation and abiotic stress in leaves and seedlings of Catharanthus roseus using geNorm, NormFinder, BestKeeper, ΔCt and RefFinder algorithms. Nine candidate reference-genes were tested for stability in C. roseus (FBOX, CACS, TIP, RSP9, EXP, EXPR, SAND, F17M5, ACT) and our study inferred that while EXP and EXPR were the most-stable, F17M5 was the lowest-stable gene in the leaf-fed samples. Among seedlings of C. roseus, F17M5 and TIP were the most, while ACT was the least-stable gene. The suitability of selected stable reference-gene pairs was demonstrated by assessing the transcript levels of the Melatonin-biosynthesis gene SNAT under same conditions. Our study is the first to comprehensively analyze the stability of commonly-used reference-genes under Melatonin-induced conditions in C. roseus.
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spelling pubmed-57991772018-02-14 Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus Sheshadri, S. A. Nishanth, M. J. Yamine, V. Simon, Bindu Sci Rep Article The role of Melatonin in influencing diverse genes in plants has gained momentum in recent years and many reports have employed qRT-PCR for their quantification. Relative quantification of gene expression relies on accurate normalization of qRT-PCR data against a stably-expressing internal reference-gene. Although researchers have been using commonly available reference-genes to assess Melatonin-induced gene expression, but to-date, there have been no attempts to validate the reference-gene stability under Melatonin-supplementation in planta. In this study, we performed stability assessment of common reference-genes under Melatonin-supplementation and abiotic stress in leaves and seedlings of Catharanthus roseus using geNorm, NormFinder, BestKeeper, ΔCt and RefFinder algorithms. Nine candidate reference-genes were tested for stability in C. roseus (FBOX, CACS, TIP, RSP9, EXP, EXPR, SAND, F17M5, ACT) and our study inferred that while EXP and EXPR were the most-stable, F17M5 was the lowest-stable gene in the leaf-fed samples. Among seedlings of C. roseus, F17M5 and TIP were the most, while ACT was the least-stable gene. The suitability of selected stable reference-gene pairs was demonstrated by assessing the transcript levels of the Melatonin-biosynthesis gene SNAT under same conditions. Our study is the first to comprehensively analyze the stability of commonly-used reference-genes under Melatonin-induced conditions in C. roseus. Nature Publishing Group UK 2018-02-05 /pmc/articles/PMC5799177/ /pubmed/29403001 http://dx.doi.org/10.1038/s41598-018-20474-2 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Sheshadri, S. A.
Nishanth, M. J.
Yamine, V.
Simon, Bindu
Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus
title Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus
title_full Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus
title_fullStr Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus
title_full_unstemmed Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus
title_short Effect of Melatonin on the stability and expression of reference genes in Catharanthus roseus
title_sort effect of melatonin on the stability and expression of reference genes in catharanthus roseus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5799177/
https://www.ncbi.nlm.nih.gov/pubmed/29403001
http://dx.doi.org/10.1038/s41598-018-20474-2
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