TY - JOUR

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

Y1 - 2016

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

U2 - 10.1016/j.bjm.2015.11.031

DO - 10.1016/j.bjm.2015.11.031

M3 - Journal article

C2 - 26887253

VL - 47

SP - 259

EP - 265

JO - Brazilian Journal of Microbiology

JF - Brazilian Journal of Microbiology

SN - 1517-8382

IS - 1

ER -