title: Evaluation of Housekeeping Genes for Quantitative Real-Time PCR Analysis of Bradysia odoriphaga (Diptera: Sciaridae).
abstract: The soil insect Bradysia odoriphaga (Diptera: Sciaridae) causes substantial damage to Chinese chive. Suitable reference genes in B. odoriphaga (Bradysia odoriphaga) have yet to be identified for normalizing target gene expression among samples by quantitative real-time PCR (qRT-PCR). This study was focused on identifying the expression stability of 12 candidate housekeeping genes in B. odoriphaga under various experiment conditions. The final stability ranking of 12 housekeeping genes was obtained with RefFinder, and the most suitable number of reference genes was analyzed by GeNorm. The results revealed that the most appropriate sets of internal controls were RPS15, RPL18, and RPS18 across developmental phases; RPS15, RPL28, and GAPDH across temperatures; RPS15 and RPL18 across pesticide treatments; RSP5, RPS18, and SDHA across photoperiods; ACTb, RPS18, and RPS15 across diets; RPS13 and RPL28 across populations; and RPS15, ACTb, and RPS18 across all samples. The use of the most suitable reference genes versus an arbitrarily selected reference gene resulted in significant differences in the analysis of a target gene expression. HSP23 in B. odoriphaga was found to be up-regulated under low temperatures. These results will contribute to the standardization of qRT-PCR and will also be valuable for further research on gene function in B. odoriphaga.
authors.affiliation: College of Agriculture, Yangtze University, Jingzhou 434025, China. shicaihua1980@126.com.
authors.forename: Caihua
authors.initials: C
authors.lastname: Shi
authors.affiliation: College of Life Science, Heilongjiang University, Harbin 150080, China. jingronghu@126.com.
authors.forename: Fengshan
authors.initials: F
authors.lastname: Yang
authors.affiliation: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China. nxy2005@126.com.
authors.forename: Xun
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA. du@uchc.edu.
authors.forename: Erxia
authors.initials: E
authors.lastname: Du
authors.affiliation: College of Agriculture, Yangtze University, Jingzhou 434025, China. nxy@yangtzeu.edu.cn.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China. wangshaoli@caas.cn.
authors.forename: Shaoli
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China. wuqingjun@caas.cn.
authors.forename: Qingjun
authors.initials: Q
authors.lastname: Wu
authors.affiliation: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China. zhangyoujun@caas.cn.
authors.forename: Youjun
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D004175:Diptera / Q000235:genetics* / Q000254:growth & development; D042963:Electron Transport Complex II / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D020043:Genes, Essential / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D007814:Larva / Q000235:genetics; D010575:Pesticides / Q000633:toxicity; D060888:Real-Time Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature
chemical_list: D000199:Actins; D019476:Insect Proteins; D010575:Pesticides; D012269:Ribosomal Proteins; D042963:Electron Transport Complex II
keywords: Bradysia odoriphaga; RefFinder; normalization; reference genes