Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in Azospirillum brasilense
Identifieur interne : 001694 ( Ncbi/Curation ); précédent : 001693; suivant : 001695Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in Azospirillum brasilense
Auteurs : Mary Mcmillan ; Lily PeregSource :
- PLoS ONE [ 1932-6203 ] ; 2014.
Abstract
Url:
DOI: 10.1371/journal.pone.0098162
PubMed: 24841066
PubMed Central: 4026395
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in <italic>Azospirillum brasilense</italic></title><author><name sortKey="Mcmillan, Mary" sort="Mcmillan, Mary" uniqKey="Mcmillan M" first="Mary" last="Mcmillan">Mary Mcmillan</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Pereg, Lily" sort="Pereg, Lily" uniqKey="Pereg L" first="Lily" last="Pereg">Lily Pereg</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">24841066</idno><idno type="pmc">4026395</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4026395</idno><idno type="RBID">PMC:4026395</idno><idno type="doi">10.1371/journal.pone.0098162</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">001202</idno><idno type="wicri:Area/Pmc/Curation">001201</idno><idno type="wicri:Area/Pmc/Checkpoint">000831</idno><idno type="wicri:Area/Ncbi/Merge">001694</idno><idno type="wicri:Area/Ncbi/Curation">001694</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in <italic>Azospirillum brasilense</italic></title><author><name sortKey="Mcmillan, Mary" sort="Mcmillan, Mary" uniqKey="Mcmillan M" first="Mary" last="Mcmillan">Mary Mcmillan</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Pereg, Lily" sort="Pereg, Lily" uniqKey="Pereg L" first="Lily" last="Pereg">Lily Pereg</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><italic>Azospirillum brasilense</italic> is a nitrogen fixing bacterium that has been shown to have various beneficial effects on plant growth and yield. Under normal conditions <italic>A. brasilense</italic> exists in a motile flagellated form, which, under starvation or stress conditions, can undergo differentiation into an encapsulated, cyst-like form. Quantitative RT-PCR can be used to analyse changes in gene expression during this differentiation process. The accuracy of quantification of mRNA levels by qRT-PCR relies on the normalisation of data against stably expressed reference genes. No suitable set of reference genes has yet been described for <italic>A. brasilense</italic>. Here we evaluated the expression of ten candidate reference genes (<italic>16S rRNA, gapB, glyA, gyrA, proC, pykA, recA, recF, rpoD, and tpiA</italic>) in wild-type and mutant <italic>A. brasilense</italic> strains under different culture conditions, including conditions that induce differentiation. Analysis with the software programs BestKeeper, NormFinder and GeNorm indicated that <italic>gyrA</italic>, <italic>glyA</italic> and <italic>recA</italic> are the most stably expressed reference genes in <italic>A. brasilense</italic>. The results also suggested that the use of two reference genes (<italic>gyrA</italic> and <italic>glyA</italic>) is sufficient for effective normalisation of qRT-PCR data.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Steenhoudt, O" uniqKey="Steenhoudt O">O Steenhoudt</name></author><author><name sortKey="Vanderleyden" uniqKey="Vanderleyden">Vanderleyden</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Herschkovitz, Y" uniqKey="Herschkovitz Y">Y Herschkovitz</name></author><author><name sortKey="Lerner, A" uniqKey="Lerner A">A Lerner</name></author><author><name sortKey="Davidov, Y" uniqKey="Davidov Y">Y Davidov</name></author><author><name sortKey="Rothballer, M" uniqKey="Rothballer M">M Rothballer</name></author><author><name sortKey="Hartmann, A" uniqKey="Hartmann A">A Hartmann</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bashan, Y" uniqKey="Bashan Y">Y 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