Cargando…
Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba
Neolamarckia cadamba is an economically-important fast-growing tree species in South China and Southeast Asia. As a prerequisite first step for future gene expression studies, we have identified and characterized a series of stable reference genes that can be used as controls for quantitative real t...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6006177/ https://www.ncbi.nlm.nih.gov/pubmed/29915368 http://dx.doi.org/10.1038/s41598-018-27633-5 |
_version_ | 1783332785509892096 |
---|---|
author | Huang, Tian Long, Jianmei Liu, Si-Wen Yang, Zi-Wei Zhu, Qi-Jin Zhao, Xiao-Lan Peng, Changcao |
author_facet | Huang, Tian Long, Jianmei Liu, Si-Wen Yang, Zi-Wei Zhu, Qi-Jin Zhao, Xiao-Lan Peng, Changcao |
author_sort | Huang, Tian |
collection | PubMed |
description | Neolamarckia cadamba is an economically-important fast-growing tree species in South China and Southeast Asia. As a prerequisite first step for future gene expression studies, we have identified and characterized a series of stable reference genes that can be used as controls for quantitative real time PCR (qRT-PCR) expression analysis in this study. The expression stability of 15 candidate reference genes in various tissues and mature leaves under different conditions was evaluated using four different algorithms, i.e., geNorm, NormFinder, BestKeeper and RefFinder. Our results showed that SAMDC was the most stable of the selected reference genes across the set of all samples, mature leaves at different photosynthetic cycles and under drought stress, whereas RPL10A had the most stable expression in various tissues. PGK and RPS25 were considered the most suitable reference for mature leaves at different developmental stages and under cold treatment, respectively. Additionally, the gene expression profiles of sucrose transporter 4 (NcSUT4), and 9‐cis‐epoxycarotenoid dioxygenase 3 (NcNCED3) were used to confirm the validity of candidate reference genes. Collectively, our study is the first report to validate the optimal reference genes for normalization under various conditions in N. cadamba and will benefit the future discovery of gene function in this species. |
format | Online Article Text |
id | pubmed-6006177 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-60061772018-06-26 Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba Huang, Tian Long, Jianmei Liu, Si-Wen Yang, Zi-Wei Zhu, Qi-Jin Zhao, Xiao-Lan Peng, Changcao Sci Rep Article Neolamarckia cadamba is an economically-important fast-growing tree species in South China and Southeast Asia. As a prerequisite first step for future gene expression studies, we have identified and characterized a series of stable reference genes that can be used as controls for quantitative real time PCR (qRT-PCR) expression analysis in this study. The expression stability of 15 candidate reference genes in various tissues and mature leaves under different conditions was evaluated using four different algorithms, i.e., geNorm, NormFinder, BestKeeper and RefFinder. Our results showed that SAMDC was the most stable of the selected reference genes across the set of all samples, mature leaves at different photosynthetic cycles and under drought stress, whereas RPL10A had the most stable expression in various tissues. PGK and RPS25 were considered the most suitable reference for mature leaves at different developmental stages and under cold treatment, respectively. Additionally, the gene expression profiles of sucrose transporter 4 (NcSUT4), and 9‐cis‐epoxycarotenoid dioxygenase 3 (NcNCED3) were used to confirm the validity of candidate reference genes. Collectively, our study is the first report to validate the optimal reference genes for normalization under various conditions in N. cadamba and will benefit the future discovery of gene function in this species. Nature Publishing Group UK 2018-06-18 /pmc/articles/PMC6006177/ /pubmed/29915368 http://dx.doi.org/10.1038/s41598-018-27633-5 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 Huang, Tian Long, Jianmei Liu, Si-Wen Yang, Zi-Wei Zhu, Qi-Jin Zhao, Xiao-Lan Peng, Changcao Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba |
title | Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba |
title_full | Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba |
title_fullStr | Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba |
title_full_unstemmed | Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba |
title_short | Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba |
title_sort | selection and validation of reference genes for mrna expression by quantitative real-time pcr analysis in neolamarckia cadamba |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6006177/ https://www.ncbi.nlm.nih.gov/pubmed/29915368 http://dx.doi.org/10.1038/s41598-018-27633-5 |
work_keys_str_mv | AT huangtian selectionandvalidationofreferencegenesformrnaexpressionbyquantitativerealtimepcranalysisinneolamarckiacadamba AT longjianmei selectionandvalidationofreferencegenesformrnaexpressionbyquantitativerealtimepcranalysisinneolamarckiacadamba AT liusiwen selectionandvalidationofreferencegenesformrnaexpressionbyquantitativerealtimepcranalysisinneolamarckiacadamba AT yangziwei selectionandvalidationofreferencegenesformrnaexpressionbyquantitativerealtimepcranalysisinneolamarckiacadamba AT zhuqijin selectionandvalidationofreferencegenesformrnaexpressionbyquantitativerealtimepcranalysisinneolamarckiacadamba AT zhaoxiaolan selectionandvalidationofreferencegenesformrnaexpressionbyquantitativerealtimepcranalysisinneolamarckiacadamba AT pengchangcao selectionandvalidationofreferencegenesformrnaexpressionbyquantitativerealtimepcranalysisinneolamarckiacadamba |