Validation of reference genes for gene expression studies in the emerald ash borer （Agrilus planipennis）

作     者：Swapna Priya Rajarapu Praveen Mamidala Omprakash Mittapalli 

作者机构：Department of Entomology The Ohio State University Ohio Agricultural and Research Development Center Wooster OH 44691 USA 

出 版 物：《Insect Science》 (昆虫科学（英文版）)

年 卷 期：2012年第19卷第1期

页      面：41-46页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0904[农学-植物保护] 0901[农学-作物学] 0836[工学-生物工程] 090402[农学-农业昆虫与害虫防治] 090102[农学-作物遗传育种] 

基　　金：Supported by a grant from the USDA APHIS / USDA Forest Service Accelerated Emerald Ash Borer Research Program State and Federal funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University 

主　　题：Agrilus planipennis BestKeeper geNorm normalization Normfinder quantitative real-time PCR reference genes 

摘      要：The Emerald ash borer （EAB, Agrilus planipennis Fairmaire） an exotic invasive insect pest has killed millions of ash trees （Fraxinus spp.） across North America and threatens billions more. We validated six A. planipennis reference genes （actin, ACT; beta tubulin, E-TUB; glyceraldehyde-3-phosphate dehydrogenase, GAPDH ; ribosomal protein, RPL7; translation elongation factor 1~, TEF-la; and ubiquitin, UBQ） using geNorm, Normfinder and BestKeeper for accurate determination of target messenger RNA levels in gene expression studies. The stability of the six reference genes was evaluated in different larval tissues, developmental stages and two treatments ofA. planipennis using quantitative real-time polymerase chain reaction. Although there was no consistent ranking observed among the reference genes across the samples, the overall analysis revealed TEF-la as the most stable reference gene. GAPDH and ACT showed least stability for all the samples studied. We conclude that TEF-I~ is the most appropriate reference gene for gene expression studies in A. planipennis. Results obtained can be applicable for transcript profiling in other invasive insect pests. Further, these validated reference genes could also serve as the basis for selection of candidate reference genes in any given insect system post-validation.