title: Selection of Reference Genes for qRT-PCR Analysis of Gene Expression in Stipa grandis during Environmental Stresses.
abstract: Stipa grandis P. Smirn. is a dominant plant species in the typical steppe of the Xilingole Plateau of Inner Mongolia. Selection of suitable reference genes for the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) is important for gene expression analysis and research into the molecular mechanisms underlying the stress responses of S. grandis. In the present study, 15 candidate reference genes (EF1 beta, ACT, GAPDH, SamDC, CUL4, CAP, SNF2, SKIP1, SKIP5, SKIP11, UBC2, UBC15, UBC17, UCH, and HERC2) were evaluated for their stability as potential reference genes for qRT-PCR under different stresses. Four algorithms were used: GeNorm, NormFinder, BestKeeper, and RefFinder. The results showed that the most stable reference genes were different under different stress conditions: EF1beta and UBC15 during drought and salt stresses; ACT and GAPDH under heat stress; SKIP5 and UBC17 under cold stress; UBC15 and HERC2 under high pH stress; UBC2 and UBC15 under wounding stress; EF1beta and UBC17 under jasmonic acid treatment; UBC15 and CUL4 under abscisic acid treatment; and HERC2 and UBC17 under salicylic acid treatment. EF1beta and HERC2 were the most suitable genes for the global analysis of all samples. Furthermore, six target genes, SgPOD, SgPAL, SgLEA, SgLOX, SgHSP90 and SgPR1, were selected to validate the most and least stable reference genes under different treatments. Our results provide guidelines for reference gene selection for more accurate qRT-PCR quantification and will promote studies of gene expression in S. grandis subjected to environmental stress.
authors.affiliation: Key Laboratory of Grassland Resources and Utilization of Ministry of Agriculture, Institute of Grassland Research, Chinese Academy of Agricultural Sciences, Hohhot, China.
authors.forename: Dongli
authors.initials: D
authors.lastname: Wan
authors.affiliation: College of Life Sciences, Inner Mongolia Agricultural University, Hohhot, China.
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Wan
authors.affiliation: College of Life Sciences, Inner Mongolia Agricultural University, Hohhot, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Yang
authors.affiliation: College of Life Sciences, Inner Mongolia Agricultural University, Hohhot, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zou
authors.affiliation: Key Laboratory of Grassland Resources and Utilization of Ministry of Agriculture, Institute of Grassland Research, Chinese Academy of Agricultural Sciences, Hohhot, China.
authors.forename: Weibo
authors.initials: W
authors.lastname: Ren
authors.affiliation: Key Laboratory of Grassland Resources and Utilization of Ministry of Agriculture, Institute of Grassland Research, Chinese Academy of Agricultural Sciences, Hohhot, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Key Laboratory of Grassland Resources and Utilization of Ministry of Agriculture, Institute of Grassland Research, Chinese Academy of Agricultural Sciences, Hohhot, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Inner Mongolia Agricultural University, Hohhot, China.
authors.forename: Ruigang
authors.initials: R
authors.lastname: Wang
authors.affiliation: Key Laboratory of Grassland Resources and Utilization of Ministry of Agriculture, Institute of Grassland Research, Chinese Academy of Agricultural Sciences, Hohhot, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Wang
authors.affiliation: Key Laboratory of Grassland Resources and Utilization of Ministry of Agriculture, Institute of Grassland Research, Chinese Academy of Agricultural Sciences, Hohhot, China.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Hou
mesh_terms: D055864:Droughts; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000235:genetics; D006358:Hot Temperature; D010940:Plant Proteins / Q000235:genetics*; D006109:Poaceae / Q000187:drug effects / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012965:Sodium Chloride / Q000633:toxicity
chemical_list: D010940:Plant Proteins; D012965:Sodium Chloride