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 expression and can add to our understanding of host-pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate 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-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.
authors.affiliation: China Jiliang University, Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, Hangzhou 310018, China. Electronic address: aspring@cjlu.edu.cn.
authors.forename: Chun
authors.initials: C
authors.lastname: Chen
authors.affiliation: China Jiliang University, Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, Hangzhou 310018, China.
authors.forename: Tingna
authors.initials: T
authors.lastname: Xie
authors.affiliation: Zhejiang Economic & Trade Polytechnic, Hangzhou 310018, China.
authors.forename: Sudan
authors.initials: S
authors.lastname: Ye
authors.affiliation: Department of Plant and Environmental Sciences, University of Copenhagen, DK 1871 Frederiksberg C, Denmark.
authors.forename: Annette Bruun
authors.initials: AB
authors.lastname: Jensen
authors.affiliation: Department of Plant and Environmental Sciences, University of Copenhagen, DK 1871 Frederiksberg C, Denmark.
authors.forename: Jørgen
authors.initials: J
authors.lastname: Eilenberg
mesh_terms: D020091:Entomophthorales / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards*; D005800:Genes, Fungal; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012339:RNA, Ribosomal, 28S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012015:Reference Standards
chemical_list: D020648:Peptide Elongation Factor 1; D012337:RNA, Ribosomal, 18S; D012339:RNA, Ribosomal, 28S
keywords: Pandora neoaphidis; Quantitative PCR; Reference genes