title: Selection of reference genes for gene expression studies in virus-infected monocots using quantitative real-time PCR.
abstract: Both genome-wide transcriptomic surveys of the mRNA expression profiles and virus-induced gene silencing-based molecular studies of target gene during virus-plant interaction involve the precise estimation of the transcript abundance. Quantitative real-time PCR (qPCR) is the most widely adopted technique for mRNA quantification. In order to obtain reliable quantification of transcripts, identification of the best reference genes forms the basis of the preliminary work. Nevertheless, the stability of internal controls in virus-infected monocots needs to be fully explored. In this work, the suitability of ten housekeeping genes (ACT, EF1α, FBOX, GAPDH, GTPB, PP2A, SAND, TUBβ, UBC18 and UK) for potential use as reference genes in qPCR were investigated in five different monocot plants (Brachypodium, barley, sorghum, wheat and maize) under infection with different viruses including Barley stripe mosaic virus (BSMV), Brome mosaic virus (BMV), Rice black-streaked dwarf virus (RBSDV) and Sugarcane mosaic virus (SCMV). By using three different algorithms, the most appropriate reference genes or their combinations were identified for different experimental sets and their effectiveness for the normalisation of expression studies were further validated by quantitative analysis of a well-studied PR-1 gene. These results facilitate the selection of desirable reference genes for more accurate gene expression studies in virus-infected monocots.
authors: State Key Laboratory of Agro-Biotechnology and Ministry of Agriculture Key Laboratory of Soil Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China.
authors: Kun
authors: K
authors: Zhang
authors: Shaofang
authors: S
authors: Niu
authors: Dianping
authors: D
authors: Di
authors: Lindan
authors: L
authors: Shi
authors: Deshui
authors: D
authors: Liu
authors: Xiuling
authors: X
authors: Cao
authors: Hongqin
authors: H
authors: Miao
authors: Xianbing
authors: X
authors: Wang
authors: Chenggui
authors: C
authors: Han
authors: Jialin
authors: J
authors: Yu
authors: Dawei
authors: D
authors: Li
authors: Yongliang
authors: Y
authors: Zhang
mesh_terms: D058431:Brachypodium / Q000235:genetics / Q000821:virology; D018506:Gene Expression Regulation, Plant / Q000235:genetics / Q000502:physiology; D001467:Hordeum / Q000235:genetics / Q000821:virology; D010940:Plant Proteins / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D045868:Sorghum / Q000235:genetics / Q000821:virology; D014908:Triticum / Q000235:genetics / Q000821:virology; D003313:Zea mays / Q000235:genetics / Q000821:virology
chemical_list: D010940:Plant Proteins
keywords: Monocots; PR-1; Reference gene; Viral infection; qPCR