Selection of reference genes for quantitative real-time PCR normalization in the plant pathogen Puccinia helianthi Schw.
Identifieur interne : 000148 ( Main/Exploration ); précédent : 000147; suivant : 000149Selection of reference genes for quantitative real-time PCR normalization in the plant pathogen Puccinia helianthi Schw.
Auteurs : Yang Song [République populaire de Chine] ; Yan Wang [République populaire de Chine] ; Dandan Guo [République populaire de Chine] ; Lan Jing [République populaire de Chine]Source :
- BMC plant biology [ 1471-2229 ] ; 2019.
Descripteurs français
- KwdFr :
- MESH :
- génétique : Régulation de l'expression des gènes végétaux.
- microbiologie : Helianthus.
- métabolisme : Helianthus.
- méthodes : RT-PCR, Réaction de polymérisation en chaine en temps réel.
- pathogénicité : Basidiomycota.
English descriptors
- KwdEn :
- MESH :
- genetics : Gene Expression Regulation, Plant.
- metabolism : Helianthus.
- methods : Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction.
- microbiology : Helianthus.
- pathogenicity : Basidiomycota.
Abstract
BACKGROUND
Real-time RT-PCR has become a common and robust technique to detect and quantify low-abundance mRNA expression and is a prefered tool when examining fungal gene expression in infected host tissues. However, correct evaluation of gene expression data requires accurate and reliable normalization against a reference transcript. Thus, the identification of reference genes with stable expression during different conditions is of paramount importance. Here, we present a study where in vitro and in planta experiments were used to validate the expression stability of reference gene candidates of Puccinia helianthi Schw., an obligate pathogen that causes rust in sunflower (Helianthus annuus).
RESULTS
Eleven reference genes of P. helianthi were validated at different growth stages. Excel-based software geNorm, BestKeeper and NormFinder were used to evaluate the reference gene transcript stabilities. Of eleven reference gene candidates tested, three were stably expressed in urediniospores, germinating growth stage and in planta. Two of these genes (UBC, EF2), encoding ubiquitin-conjugating enzyme and elongation factor 2, proved to be the most stable set of reference genes under the experimental conditions used.
CONCLUSION
We found that UBC and EF2 are suitable candidates for for the standardization of gene expression studies in the plant pathogen P. helianthi and potentially other related pathogens.
DOI: 10.1186/s12870-019-1629-x
PubMed: 30634896
PubMed Central: PMC6329156
Affiliations:
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Schw.</title><author><name sortKey="Song, Yang" sort="Song, Yang" uniqKey="Song Y" first="Yang" last="Song">Yang Song</name><affiliation wicri:level="1"><nlm:affiliation>College of Agronomy, Inner Mongolia Agricultural University, Hohhot, 010019, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Agronomy, Inner Mongolia Agricultural University, Hohhot, 010019</wicri:regionArea><wicri:noRegion>010019</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Yan" sort="Wang, Yan" uniqKey="Wang Y" first="Yan" last="Wang">Yan Wang</name><affiliation wicri:level="1"><nlm:affiliation>College of Agronomy, Inner Mongolia Agricultural University, Hohhot, 010019, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Agronomy, Inner Mongolia Agricultural University, Hohhot, 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Hohhot, 010019</wicri:regionArea><wicri:noRegion>010019</wicri:noRegion></affiliation></author></analytic><series><title level="j">BMC plant biology</title><idno type="eISSN">1471-2229</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Basidiomycota (pathogenicity)</term><term>Gene Expression Regulation, Plant (genetics)</term><term>Helianthus (metabolism)</term><term>Helianthus (microbiology)</term><term>Real-Time Polymerase Chain Reaction (methods)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Basidiomycota (pathogénicité)</term><term>Helianthus (microbiologie)</term><term>Helianthus (métabolisme)</term><term>RT-PCR (méthodes)</term><term>Réaction de polymérisation en chaine en temps réel (méthodes)</term><term>Régulation de l'expression des gènes végétaux (génétique)</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Gene Expression Regulation, Plant</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Régulation de l'expression des gènes végétaux</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Helianthus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Real-Time Polymerase Chain Reaction</term><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="microbiologie" xml:lang="fr"><term>Helianthus</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Helianthus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Helianthus</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>RT-PCR</term><term>Réaction de polymérisation en chaine en temps réel</term></keywords><keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>Basidiomycota</term></keywords><keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr"><term>Basidiomycota</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b></p><p>Real-time RT-PCR has become a common and robust technique to detect and quantify low-abundance mRNA expression and is a prefered tool when examining fungal gene expression in infected host tissues. However, correct evaluation of gene expression data requires accurate and reliable normalization against a reference transcript. Thus, the identification of reference genes with stable expression during different conditions is of paramount importance. Here, we present a study where in vitro and in planta experiments were used to validate the expression stability of reference gene candidates of Puccinia helianthi Schw., an obligate pathogen that causes rust in sunflower (Helianthus annuus).</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>Eleven reference genes of P. helianthi were validated at different growth stages. Excel-based software geNorm, BestKeeper and NormFinder were used to evaluate the reference gene transcript stabilities. Of eleven reference gene candidates tested, three were stably expressed in urediniospores, germinating growth stage and in planta. Two of these genes (UBC, EF2), encoding ubiquitin-conjugating enzyme and elongation factor 2, proved to be the most stable set of reference genes under the experimental conditions used.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSION</b></p><p>We found that UBC and EF2 are suitable candidates for for the standardization of gene expression studies in the plant pathogen P. helianthi and potentially other related pathogens.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">30634896</PMID><DateCompleted><Year>2019</Year><Month>03</Month><Day>04</Day></DateCompleted><DateRevised><Year>2020</Year><Month>02</Month><Day>25</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1471-2229</ISSN><JournalIssue CitedMedium="Internet"><Volume>19</Volume><Issue>1</Issue><PubDate><Year>2019</Year><Month>Jan</Month><Day>11</Day></PubDate></JournalIssue><Title>BMC plant biology</Title><ISOAbbreviation>BMC Plant Biol</ISOAbbreviation></Journal><ArticleTitle>Selection of reference genes for quantitative real-time PCR normalization in the plant pathogen Puccinia helianthi Schw.</ArticleTitle><Pagination><MedlinePgn>20</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1186/s12870-019-1629-x</ELocationID><Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">Real-time RT-PCR has become a common and robust technique to detect and quantify low-abundance mRNA expression and is a prefered tool when examining fungal gene expression in infected host tissues. However, correct evaluation of gene expression data requires accurate and reliable normalization against a reference transcript. Thus, the identification of reference genes with stable expression during different conditions is of paramount importance. Here, we present a study where in vitro and in planta experiments were used to validate the expression stability of reference gene candidates of Puccinia helianthi Schw., an obligate pathogen that causes rust in sunflower (Helianthus annuus).</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">Eleven reference genes of P. helianthi were validated at different growth stages. Excel-based software geNorm, BestKeeper and NormFinder were used to evaluate the reference gene transcript stabilities. Of eleven reference gene candidates tested, three were stably expressed in urediniospores, germinating growth stage and in planta. Two of these genes (UBC, EF2), encoding ubiquitin-conjugating enzyme and elongation factor 2, proved to be the most stable set of reference genes under the experimental conditions used.</AbstractText><AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">We found that UBC and EF2 are suitable candidates for for the standardization of gene expression studies in the plant pathogen P. helianthi and potentially other related pathogens.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Song</LastName><ForeName>Yang</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>College of Agronomy, Inner Mongolia Agricultural University, Hohhot, 010019, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Yan</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>College of Agronomy, Inner Mongolia Agricultural University, Hohhot, 010019, China.</Affiliation></AffiliationInfo></Author><Author 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