About: The choice of reference gene set for assessing gene expression in barley (Hordeum vulgare L.) under low temperature and drought stress     Goto   Sponge   NotDistinct   Permalink

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Description
  • Drought and low temperature are the two most significant causes of abiotic stress in agricultural crops and, therefore, they pose considerable challenges in plant science. Hence, it is crucial to study response mechanisms and to select genes for identification signaling pathways that lead from stimulus to response. The assessment of gene expression is often attempted using real-time RT-PCR (qRT-PCR), a technique which requires a careful choice of reference gene(s) for normalization purpose. Here, we report a comparison of 13 potential reference genes for studying gene expression in the leaf and crown of barley seedlings subjected to low temperature or drought stress. All three currently available software packages designed to identify reference genes from qRT-PCR data (GeNorm, NormFinder and BestKeeper) were used to identify informative sets of up to three reference genes. Interestingly, the data obtained from the separate treatment of leaf and crown have led to the recommendations that HSP70 and S-AMD (and possibly HSP90) to be used as the reference genes for low-temperature stressed leaves, HSP90 and EF1 alpha for low-temperature stressed crowns, cyclophilin and ADP-RF (and possibly ACT) for drought-stressed leaves, and EF1 alpha and S-AMD for drought-stressed crowns. Our results have demonstrated that the gene expression can be highly tissue- or organ-specific in barley and have confirmed that reference gene choice is essential in qRT-PCR. The findings can also serve as guidelines for the selection of reference genes under different stress conditions and lay foundation for more accurate and widespread use of qRT-PCR in barley gene analysis.
  • Drought and low temperature are the two most significant causes of abiotic stress in agricultural crops and, therefore, they pose considerable challenges in plant science. Hence, it is crucial to study response mechanisms and to select genes for identification signaling pathways that lead from stimulus to response. The assessment of gene expression is often attempted using real-time RT-PCR (qRT-PCR), a technique which requires a careful choice of reference gene(s) for normalization purpose. Here, we report a comparison of 13 potential reference genes for studying gene expression in the leaf and crown of barley seedlings subjected to low temperature or drought stress. All three currently available software packages designed to identify reference genes from qRT-PCR data (GeNorm, NormFinder and BestKeeper) were used to identify informative sets of up to three reference genes. Interestingly, the data obtained from the separate treatment of leaf and crown have led to the recommendations that HSP70 and S-AMD (and possibly HSP90) to be used as the reference genes for low-temperature stressed leaves, HSP90 and EF1 alpha for low-temperature stressed crowns, cyclophilin and ADP-RF (and possibly ACT) for drought-stressed leaves, and EF1 alpha and S-AMD for drought-stressed crowns. Our results have demonstrated that the gene expression can be highly tissue- or organ-specific in barley and have confirmed that reference gene choice is essential in qRT-PCR. The findings can also serve as guidelines for the selection of reference genes under different stress conditions and lay foundation for more accurate and widespread use of qRT-PCR in barley gene analysis. (en)
Title
  • The choice of reference gene set for assessing gene expression in barley (Hordeum vulgare L.) under low temperature and drought stress
  • The choice of reference gene set for assessing gene expression in barley (Hordeum vulgare L.) under low temperature and drought stress (en)
skos:prefLabel
  • The choice of reference gene set for assessing gene expression in barley (Hordeum vulgare L.) under low temperature and drought stress
  • The choice of reference gene set for assessing gene expression in barley (Hordeum vulgare L.) under low temperature and drought stress (en)
skos:notation
  • RIV/00216208:11310/13:10195352!RIV14-MZE-11310___
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • I, P(OC09032), P(QH81287), S, Z(MZE0002700604)
http://linked.open...iv/cisloPeriodika
  • 11
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 65328
http://linked.open...ai/riv/idVysledku
  • RIV/00216208:11310/13:10195352
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • Real-time RT PCR; Normalization; Low temperature; Leaves; Drought; Crowns (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • DE - Spolková republika Německo
http://linked.open...ontrolniKodProRIV
  • [081DB6B6DF36]
http://linked.open...i/riv/nazevZdroje
  • Molecular Genetics and Genomics
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 288
http://linked.open...iv/tvurceVysledku
  • Janská, Anna
  • Svoboda, Pavel
  • Hodek, Jan
  • Milella, Luigi
  • Ovesna, Jaroslava
  • Prasil, Ilja Tom
  • Vlasakova, Eva
  • Zamecnik, Jiří
http://linked.open...ain/vavai/riv/wos
  • 000326263900008
http://linked.open...n/vavai/riv/zamer
issn
  • 1617-4615
number of pages
http://bibframe.org/vocab/doi
  • 10.1007/s00438-013-0774-4
http://localhost/t...ganizacniJednotka
  • 11310
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