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Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata

The desert pioneer plant Stipagrostis pennata plays an important role in sand fixation, wind prevention, and desert ecosystem recovery. An absence of reference genes greatly limits investigations into the regulatory mechanism by which S. pennata adapts to adverse desert environments at the molecular...

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Autores principales: Li, Rong, Cui, Kaiwen, Xie, Quanliang, Xie, Shuangquan, Chen, Xifeng, Zhuo, Lu, Cao, Aiping, Shen, Haitao, Jin, Xiang, Wang, Fei, Li, Hongbin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8571334/
https://www.ncbi.nlm.nih.gov/pubmed/34741052
http://dx.doi.org/10.1038/s41598-021-00833-2
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author Li, Rong
Cui, Kaiwen
Xie, Quanliang
Xie, Shuangquan
Chen, Xifeng
Zhuo, Lu
Cao, Aiping
Shen, Haitao
Jin, Xiang
Wang, Fei
Li, Hongbin
author_facet Li, Rong
Cui, Kaiwen
Xie, Quanliang
Xie, Shuangquan
Chen, Xifeng
Zhuo, Lu
Cao, Aiping
Shen, Haitao
Jin, Xiang
Wang, Fei
Li, Hongbin
author_sort Li, Rong
collection PubMed
description The desert pioneer plant Stipagrostis pennata plays an important role in sand fixation, wind prevention, and desert ecosystem recovery. An absence of reference genes greatly limits investigations into the regulatory mechanism by which S. pennata adapts to adverse desert environments at the molecular and genetic levels. In this study, eight candidate reference genes were identified from rhizosheath development transcriptome data from S. pennata, and their expression stability in the rhizosheaths at different development stages, in a variety of plant tissues, and under drought stress was evaluated using four procedures, including geNorm, NormFinder, BestKeeper, and RefFinder. The results showed that GAPDH and elF were the most stable reference genes under drought stress and in rhizosheath development, and ARP6 and ALDH were relatively stable in all plant tissues. In addition, elF was the most suitable reference gene for all treatments. Analysis of the consistency between the reverse transcription-quantitative PCR (RT-qPCR) and RNA sequencing data showed that the identified elF and GAPDH reference genes were stable during rhizosheath development. These results provide reliable reference genes for assuring the accuracy of RT-qPCR and offer a foundation for further investigations into the genetic responses of S. pennata to abiotic stress.
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spelling pubmed-85713342021-11-09 Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata Li, Rong Cui, Kaiwen Xie, Quanliang Xie, Shuangquan Chen, Xifeng Zhuo, Lu Cao, Aiping Shen, Haitao Jin, Xiang Wang, Fei Li, Hongbin Sci Rep Article The desert pioneer plant Stipagrostis pennata plays an important role in sand fixation, wind prevention, and desert ecosystem recovery. An absence of reference genes greatly limits investigations into the regulatory mechanism by which S. pennata adapts to adverse desert environments at the molecular and genetic levels. In this study, eight candidate reference genes were identified from rhizosheath development transcriptome data from S. pennata, and their expression stability in the rhizosheaths at different development stages, in a variety of plant tissues, and under drought stress was evaluated using four procedures, including geNorm, NormFinder, BestKeeper, and RefFinder. The results showed that GAPDH and elF were the most stable reference genes under drought stress and in rhizosheath development, and ARP6 and ALDH were relatively stable in all plant tissues. In addition, elF was the most suitable reference gene for all treatments. Analysis of the consistency between the reverse transcription-quantitative PCR (RT-qPCR) and RNA sequencing data showed that the identified elF and GAPDH reference genes were stable during rhizosheath development. These results provide reliable reference genes for assuring the accuracy of RT-qPCR and offer a foundation for further investigations into the genetic responses of S. pennata to abiotic stress. Nature Publishing Group UK 2021-11-05 /pmc/articles/PMC8571334/ /pubmed/34741052 http://dx.doi.org/10.1038/s41598-021-00833-2 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Li, Rong
Cui, Kaiwen
Xie, Quanliang
Xie, Shuangquan
Chen, Xifeng
Zhuo, Lu
Cao, Aiping
Shen, Haitao
Jin, Xiang
Wang, Fei
Li, Hongbin
Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata
title Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata
title_full Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata
title_fullStr Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata
title_full_unstemmed Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata
title_short Selection of the reference genes for quantitative gene expression by RT-qPCR in the desert plant Stipagrostis pennata
title_sort selection of the reference genes for quantitative gene expression by rt-qpcr in the desert plant stipagrostis pennata
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8571334/
https://www.ncbi.nlm.nih.gov/pubmed/34741052
http://dx.doi.org/10.1038/s41598-021-00833-2
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