Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis

Chun Chen; Tingna Xie; Sudan Ye; Annette Bruun Jensen; Jørgen Eilenberg

doi:10.1016/j.bjm.2015.11.031

Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis

Chun Chen, Tingna Xie, Sudan Ye, Annette Bruun Jensen, Jørgen Eilenberg

SOBI

12 Citationer (Scopus)

42 Downloads (Pure)

Abstract

The selection of suitable reference genes is crucial for accurate quantification of gene expres-sion and can add to our understanding of host–pathogen interactions. To identify suitablereference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expressionof three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongationfactor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reac-tion at different developmental stages (conidia, conidia with germ tubes, short hyphae andelongated hyphae), and under different nutritional conditions. We calculated the expressionstability of candidate reference genes using four algorithms including geNorm, NormFinder,BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking ofcandidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414)and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCRanalysis of gene expression under all conditions. These results will support further studieson gene expression in P. neoaphidis

Originalsprog	Engelsk
Tidsskrift	Brazilian Journal of Microbiology
Vol/bind	47
Udgave nummer	1
Sider (fra-til)	259-265
Antal sider	7
ISSN	1517-8382
DOI	https://doi.org/10.1016/j.bjm.2015.11.031
Status	Udgivet - 1 jan. 2016

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10.1016/j.bjm.2015.11.031Licens: CC BY-NC-ND

1-s2.0-S1517838215000349-mainForlagets udgivne version, 962 KBLicens: CC BY-NC-ND

Citationsformater

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title = "Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis",

abstract = "The selection of suitable reference genes is crucial for accurate quantification of gene expres-sion and can add to our understanding of host–pathogen interactions. To identify suitablereference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expressionof three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongationfactor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reac-tion at different developmental stages (conidia, conidia with germ tubes, short hyphae andelongated hyphae), and under different nutritional conditions. We calculated the expressionstability of candidate reference genes using four algorithms including geNorm, NormFinder,BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking ofcandidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414)and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCRanalysis of gene expression under all conditions. These results will support further studieson gene expression in P. neoaphidis",

author = "Chun Chen and Tingna Xie and Sudan Ye and Jensen, {Annette Bruun} and J{\o}rgen Eilenberg",

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T1 - Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis

AU - Chen, Chun

AU - Xie, Tingna

AU - Ye, Sudan

AU - Jensen, Annette Bruun

AU - Eilenberg, Jørgen

PY - 2016/1/1

Y1 - 2016/1/1

N2 - The selection of suitable reference genes is crucial for accurate quantification of gene expres-sion and can add to our understanding of host–pathogen interactions. To identify suitablereference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expressionof three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongationfactor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reac-tion at different developmental stages (conidia, conidia with germ tubes, short hyphae andelongated hyphae), and under different nutritional conditions. We calculated the expressionstability of candidate reference genes using four algorithms including geNorm, NormFinder,BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking ofcandidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414)and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCRanalysis of gene expression under all conditions. These results will support further studieson gene expression in P. neoaphidis

AB - The selection of suitable reference genes is crucial for accurate quantification of gene expres-sion and can add to our understanding of host–pathogen interactions. To identify suitablereference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expressionof three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongationfactor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reac-tion at different developmental stages (conidia, conidia with germ tubes, short hyphae andelongated hyphae), and under different nutritional conditions. We calculated the expressionstability of candidate reference genes using four algorithms including geNorm, NormFinder,BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking ofcandidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414)and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCRanalysis of gene expression under all conditions. These results will support further studieson gene expression in P. neoaphidis

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DO - 10.1016/j.bjm.2015.11.031

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JO - Brazilian Journal of Microbiology

JF - Brazilian Journal of Microbiology

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