Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration
Syntrichia caninervis is the dominant bryophyte of the biological soil crusts found in the Gurbantunggut desert. The extreme desert environment is characterized by prolonged drought, temperature extremes, high radiation and frequent cycles of hydration and dehydration. S. caninervis is an ideal org...
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doaj-88d0b9e63fb54f31b977cfa1ad60e5b02020-11-24T23:02:45ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2015-02-01610.3389/fpls.2015.00038123059Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydrationXiaoshuang eLi0Daoyuan eZhang1Haiyan eLi2Bei eGao3Honglan eYang4Yuanming eZhang5Andrew eWood6Xinjiang Institute of Ecology and Geography, Chinese Academy of SciencesXinjiang Institute of Ecology and Geography, Chinese Academy of SciencesXinjiang Institute of Ecology and Geography, Chinese Academy of SciencesXinjiang Institute of Ecology and Geography, Chinese Academy of SciencesXinjiang Institute of Ecology and Geography, Chinese Academy of SciencesXinjiang Institute of Ecology and Geography, Chinese Academy of SciencesSouthern Illinois UniversitySyntrichia caninervis is the dominant bryophyte of the biological soil crusts found in the Gurbantunggut desert. The extreme desert environment is characterized by prolonged drought, temperature extremes, high radiation and frequent cycles of hydration and dehydration. S. caninervis is an ideal organism for the identification and characterization of genes related to abiotic stress tolerance. RT-qPCR expression analysis is a powerful analytical technique that requires the use of stable reference genes. Using available S. caninervis transcriptome data, we selected 15 candidate reference genes and analyzed their relative expression stabilities in S. caninervis gametophores exposed to a range of abiotic stresses or a hydration-desiccation-rehydration cycle. The programs geNorm, NormFinder, and RefFinder were used to assess and rank the expression stability of the 15 candidate genes. The stability ranking results of reference genes under each specific experimental condition showed high consistency using different algorithms. For abiotic stress treatments, the combination of two genes (α-TUB2 and CDPK) were sufficient for accurate normalization. For the hydration-desiccation-rehydration process, the combination of two genes (α-TUB1 and CDPK) were sufficient for accurate normalization. 18S was among the least stable genes in all of the experimental sets and was unsuitable as reference gene in S. caninervis. This is the first systematic investigation and comparison of reference gene selection for RT-qPCR work in S. caninervis. This research will facilitate gene expression studies in S. caninervis, related moss species from the Syntrichia complex and other mosses.http://journal.frontiersin.org/Journal/10.3389/fpls.2015.00038/fullreference genequantitative real-time PCRgeNormNormFinderSyntrichia caninervisRefFinder |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Xiaoshuang eLi Daoyuan eZhang Haiyan eLi Bei eGao Honglan eYang Yuanming eZhang Andrew eWood |
spellingShingle |
Xiaoshuang eLi Daoyuan eZhang Haiyan eLi Bei eGao Honglan eYang Yuanming eZhang Andrew eWood Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration Frontiers in Plant Science reference gene quantitative real-time PCR geNorm NormFinder Syntrichia caninervis RefFinder |
author_facet |
Xiaoshuang eLi Daoyuan eZhang Haiyan eLi Bei eGao Honglan eYang Yuanming eZhang Andrew eWood |
author_sort |
Xiaoshuang eLi |
title |
Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration |
title_short |
Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration |
title_full |
Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration |
title_fullStr |
Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration |
title_full_unstemmed |
Characterization of reference genes for RT-qPCR in the desert moss Syntrichia caninervis in response to abiotic stress and desiccation/rehydration |
title_sort |
characterization of reference genes for rt-qpcr in the desert moss syntrichia caninervis in response to abiotic stress and desiccation/rehydration |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Plant Science |
issn |
1664-462X |
publishDate |
2015-02-01 |
description |
Syntrichia caninervis is the dominant bryophyte of the biological soil crusts found in the Gurbantunggut desert. The extreme desert environment is characterized by prolonged drought, temperature extremes, high radiation and frequent cycles of hydration and dehydration. S. caninervis is an ideal organism for the identification and characterization of genes related to abiotic stress tolerance. RT-qPCR expression analysis is a powerful analytical technique that requires the use of stable reference genes. Using available S. caninervis transcriptome data, we selected 15 candidate reference genes and analyzed their relative expression stabilities in S. caninervis gametophores exposed to a range of abiotic stresses or a hydration-desiccation-rehydration cycle. The programs geNorm, NormFinder, and RefFinder were used to assess and rank the expression stability of the 15 candidate genes. The stability ranking results of reference genes under each specific experimental condition showed high consistency using different algorithms. For abiotic stress treatments, the combination of two genes (α-TUB2 and CDPK) were sufficient for accurate normalization. For the hydration-desiccation-rehydration process, the combination of two genes (α-TUB1 and CDPK) were sufficient for accurate normalization. 18S was among the least stable genes in all of the experimental sets and was unsuitable as reference gene in S. caninervis. This is the first systematic investigation and comparison of reference gene selection for RT-qPCR work in S. caninervis. This research will facilitate gene expression studies in S. caninervis, related moss species from the Syntrichia complex and other mosses. |
topic |
reference gene quantitative real-time PCR geNorm NormFinder Syntrichia caninervis RefFinder |
url |
http://journal.frontiersin.org/Journal/10.3389/fpls.2015.00038/full |
work_keys_str_mv |
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