title: Identification of novel and robust internal control genes from Volvariella volvacea that are suitable for RT-qPCR in filamentous fungi.
abstract: The selection of appropriate internal control genes (ICGs) is a crucial step in the normalization of real-time quantitative PCR (RT-qPCR) data. Housekeeping genes are habitually selected for this purpose, despite accumulating evidence on their instability. We screened for novel, robust ICGs in the mushroom forming fungus Volvariella volvacea. Nine commonly used and five newly selected ICGs were evaluated for expression stability using RT-qPCR data in eight different stages of the life cycle of V. volvacea. Three different algorithms consistently determined that three novel ICGs (SPRYp, Ras and Vps26) exhibited the highest expression stability in V. volvacea. Subsequent analysis of ICGs in twenty-four expression profiles from nine filamentous fungi revealed that Ras was the most stable ICG amongst the Basidiomycetous samples, followed by SPRYp, Vps26 and ACTB. Vps26 was expressed most stably within the analyzed data of Ascomycetes, followed by HH3 and β-TUB. No ICG was universally stable for all fungal species, or for all experimental conditions within a species. Ultimately, the choice of an ICG will depend on a specific set of experiments. This study provides novel, robust ICGs for Basidiomycetes and Ascomycetes. Together with the presented guiding principles, this enables the efficient selection of suitable ICGs for RT-qPCR.
authors.affiliation: College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.|Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Yongxin
authors.initials: Y
authors.lastname: Tao
authors.affiliation: Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Arend Frans
authors.initials: AF
authors.lastname: van Peer
authors.affiliation: College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.|Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Qianhui
authors.initials: Q
authors.lastname: Huang
authors.affiliation: College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.|Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Xie
authors.affiliation: Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.|College of Food Science, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.|Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Mycological Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China.
authors.forename: Baogui
authors.initials: B
authors.lastname: Xie
mesh_terms: D005656:Fungal Proteins / Q000235:genetics*; D005658:Fungi / Q000235:genetics*; D020043:Genes, Essential / Q000235:genetics*; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D055367:Volvariella / Q000235:genetics
chemical_list: D005656:Fungal Proteins