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Statements

Subject Item
n2:RIV%2F00216208%3A11310%2F13%3A10195352%21RIV14-MZE-11310___
rdf:type
n5:Vysledek skos:Concept
rdfs:seeAlso
http://dx.doi.org/10.1007/s00438-013-0774-4
dcterms: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.
dcterms: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
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
skos:notation
RIV/00216208:11310/13:10195352!RIV14-MZE-11310___
n5:predkladatel
n10:orjk%3A11310
n3:aktivita
n19:Z n19:S n19:P n19:I
n3:aktivity
I, P(OC09032), P(QH81287), S, Z(MZE0002700604)
n3:cisloPeriodika
11
n3:dodaniDat
n13:2014
n3:domaciTvurceVysledku
n9:4949811
n3:druhVysledku
n21:J
n3:duvernostUdaju
n16:S
n3:entitaPredkladatele
n11:predkladatel
n3:idSjednocenehoVysledku
65328
n3:idVysledku
RIV/00216208:11310/13:10195352
n3:jazykVysledku
n6:eng
n3:klicovaSlova
Real-time RT PCR; Normalization; Low temperature; Leaves; Drought; Crowns
n3:klicoveSlovo
n8:Leaves n8:Normalization n8:Real-time%20RT%20PCR n8:Low%20temperature n8:Crowns n8:Drought
n3:kodStatuVydavatele
DE - Spolková republika Německo
n3:kontrolniKodProRIV
[081DB6B6DF36]
n3:nazevZdroje
Molecular Genetics and Genomics
n3:obor
n22:ED
n3:pocetDomacichTvurcuVysledku
1
n3:pocetTvurcuVysledku
8
n3:projekt
n7:QH81287 n7:OC09032
n3:rokUplatneniVysledku
n13:2013
n3:svazekPeriodika
288
n3:tvurceVysledku
Prasil, Ilja Tom Svoboda, Pavel Vlasakova, Eva Zamecnik, Jiří Janská, Anna Hodek, Jan Ovesna, Jaroslava Milella, Luigi
n3:wos
000326263900008
n3:zamer
n15:MZE0002700604
s:issn
1617-4615
s:numberOfPages
11
n14:doi
10.1007/s00438-013-0774-4
n17:organizacniJednotka
11310