Screening and Validation of Stable Reference Genes for qRT-PCR Analysis in <i>Epicauta gorhami</i> (Coleoptera: Meloidae)
Guofeng Yang,
Xuetao Yu,
Yan Zhang,
Jinhua Luo,
Xiaofei Li,
Li Zhu,
Huanhuan Zhang,
Lin Jin,
Gang Wu,
Xiaohong Yan,
Chenhui Shen
Affiliations
Guofeng Yang
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Xuetao Yu
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Yan Zhang
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Jinhua Luo
Enshi Tujia and Miao Autonomous Prefecture Academy of Agricultural Sciences, Enshi 445000, China
Xiaofei Li
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Li Zhu
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Huanhuan Zhang
Institute of Vegetable, Tibet Academy of Agricultural and Animal Husbandry Sciences, Lhasa 850032, China
Lin Jin
Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Gang Wu
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Xiaohong Yan
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Chenhui Shen
Key Laboratory of Agricultural Genetically Modified Organisms Traceability, Oil Crops Research Institute of Chinese Academy of Agricultural Science/Supervision and Test Center (Wuhan) for Plant Ecological Environment Safety, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
Epicauta gorhami is a hypermetamorphic insect that mainly forage soybeans during the adult stage. However, the lack of appropriate references hinders our studying of the gene function in E. gorhami. In this study, referring to five computational tools (Ct value, geNorm, NormFinder, BestKeeper and RefFinder), the stability of 10 housekeeping genes (GAPDH, ACT, RPL4, RPL27, α-TUB, RPS18, EF1α, RPS28, RPL13 and SOD) was assessed by qRT-PCR under three different conditions (adult ages, tissues/organs and temperatures). The findings suggested that SOD and RPS18 were the most ideal references for examine gene transcripts among diverse adult ages and at various temperatures; a pair of RPS18 and RPS28 was the most reliable genes to assess gene expressions in diverse adult tissues. Finally, the relative expression levels of EgUAP were computed after normalization RPS18 and RPS28 with across diverse adult tissues. As expected, EgUAP expression was abundant in the foregut, trachea and antenna and scarce in the midgut, hindgut and epidermis. These findings will lay a solid foundation for analyzing the gene expression of E. gorhami.
