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

Section for Organismal Biology

12 Citations (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

Original language	English
Journal	Brazilian Journal of Microbiology
Volume	47
Issue number	1
Pages (from-to)	259-265
Number of pages	7
ISSN	1517-8382
DOIs	https://doi.org/10.1016/j.bjm.2015.11.031
Publication status	Published - 1 Jan 2016

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Cite this

@article{961db5548f354be0966e54c2e2e47622,

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",

year = "2016",

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language = "English",

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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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