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Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo

Dendrobium officinale Kimu et Migo has increased many researchers’ interest for its high medical and horticultural values and the molecular mechanism of its protocorm development remains unclear. In this study, 19 genes from 26 most stably expressed genes in whole transcriptome of protocorms and 5 h...

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Autores principales: An, Hongqiang, Zhu, Qiankun, Pei, Wei, Fan, Jing, Liang, Yi, Cui, Yihui, Lv, Nan, Wang, Wanjun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5096709/
https://www.ncbi.nlm.nih.gov/pubmed/27814359
http://dx.doi.org/10.1371/journal.pone.0163478
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author An, Hongqiang
Zhu, Qiankun
Pei, Wei
Fan, Jing
Liang, Yi
Cui, Yihui
Lv, Nan
Wang, Wanjun
author_facet An, Hongqiang
Zhu, Qiankun
Pei, Wei
Fan, Jing
Liang, Yi
Cui, Yihui
Lv, Nan
Wang, Wanjun
author_sort An, Hongqiang
collection PubMed
description Dendrobium officinale Kimu et Migo has increased many researchers’ interest for its high medical and horticultural values and the molecular mechanism of its protocorm development remains unclear. In this study, 19 genes from 26 most stably expressed genes in whole transcriptome of protocorms and 5 housekeeping genes were used as candidate reference genes and screened with 4 application softwares (geNorm, NormFinder, BestKeeper and RefFinder). The results showed that a few reference genes could effectively normalize expression level of specific genes in protocorm development and the optimal top 2 reference genes were ASS and APH1L. Meanwhile, validation of GNOM, AP2 and temperature induced gene (TIL) for normalization demonstrates the usefulness of the validated candidate reference genes. The expression profiles of these genes varied under protocorms and temperature stress according to the stablest and unstablest reference genes, which proved the importance of the choice of appropriate reference genes. The first systematic evaluation of stably expressed genes will be very useful in the future analysis of specific genes expression in D. officinale.
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spelling pubmed-50967092016-11-18 Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo An, Hongqiang Zhu, Qiankun Pei, Wei Fan, Jing Liang, Yi Cui, Yihui Lv, Nan Wang, Wanjun PLoS One Research Article Dendrobium officinale Kimu et Migo has increased many researchers’ interest for its high medical and horticultural values and the molecular mechanism of its protocorm development remains unclear. In this study, 19 genes from 26 most stably expressed genes in whole transcriptome of protocorms and 5 housekeeping genes were used as candidate reference genes and screened with 4 application softwares (geNorm, NormFinder, BestKeeper and RefFinder). The results showed that a few reference genes could effectively normalize expression level of specific genes in protocorm development and the optimal top 2 reference genes were ASS and APH1L. Meanwhile, validation of GNOM, AP2 and temperature induced gene (TIL) for normalization demonstrates the usefulness of the validated candidate reference genes. The expression profiles of these genes varied under protocorms and temperature stress according to the stablest and unstablest reference genes, which proved the importance of the choice of appropriate reference genes. The first systematic evaluation of stably expressed genes will be very useful in the future analysis of specific genes expression in D. officinale. Public Library of Science 2016-11-04 /pmc/articles/PMC5096709/ /pubmed/27814359 http://dx.doi.org/10.1371/journal.pone.0163478 Text en © 2016 An 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, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
An, Hongqiang
Zhu, Qiankun
Pei, Wei
Fan, Jing
Liang, Yi
Cui, Yihui
Lv, Nan
Wang, Wanjun
Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo
title Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo
title_full Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo
title_fullStr Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo
title_full_unstemmed Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo
title_short Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo
title_sort whole-transcriptome selection and evaluation of internal reference genes for expression analysis in protocorm development of dendrobium officinale kimura et migo
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5096709/
https://www.ncbi.nlm.nih.gov/pubmed/27814359
http://dx.doi.org/10.1371/journal.pone.0163478
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