title: Selection of suitable reference genes for qPCR normalization under abiotic stresses in Oenanthe javanica (BI.) DC.
abstract: Accurate normalization of gene expression data is an absolute prerequisite to obtain reliable results in qPCR analysis. Oenanthe javanica, an aquatic perennial herb, belongs to the Oenanthe genus in Apiaceae family, with known medicinal properties. In the current study, O. javanica was subjected to hormone stimuli (gibberellin, salicylic acid, methyl jasmonate, and abscisic acid) and abiotic stresses (heat, cold, salt, and drought), and the expression of nine candidate reference genes (eIF-4α, ACT7, TIP41, GAPDH, SAND, EF-1α, PP2A, TBP, and TUB) was evaluated. Stability of the genes was assessed using geNorm, NormFinder and BestKeeper. All the genes presented distinct expression profiles under the experimental conditions analyzed. Under abiotic stress conditions, ACT7 and PP2A genes displayed the maximum stability; PP2A and SAND were the most stable genes under hormone stimuli. Even though PP2A gene was most stable across all the samples, individual analysis revealed changes in expression profile. To further validate the suitability of the reference genes identified in this study, the expression level of M6PR gene under salt treatment was studied. Based on our data, we propose that it is essential to normalize the target gene expression with specific reference genes under different experimental conditions for most accurate results. To our knowledge, this is the first systematic analysis for reference genes under abiotic stress and hormone stimuli conditions in O. javanica. This will be beneficial for future studies on O. javanica and other plants in Apiaceae family at molecular level.
authors.affiliation: State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China of Ministry of Agriculture, College of Horticulture, Nanjing Agricultural University, Nanjing, People's Republic China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China of Ministry of Agriculture, College of Horticulture, Nanjing Agricultural University, Nanjing, People's Republic China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China of Ministry of Agriculture, College of Horticulture, Nanjing Agricultural University, Nanjing, People's Republic China.
authors.forename: Meng-Yao
authors.initials: MY
authors.lastname: Li
authors.affiliation: State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China of Ministry of Agriculture, College of Horticulture, Nanjing Agricultural University, Nanjing, People's Republic China.
authors.forename: Jing
authors.initials: J
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China of Ministry of Agriculture, College of Horticulture, Nanjing Agricultural University, Nanjing, People's Republic China.
authors.forename: Guo-Fei
authors.initials: GF
authors.lastname: Tan
authors.affiliation: State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China of Ministry of Agriculture, College of Horticulture, Nanjing Agricultural University, Nanjing, People's Republic China.
authors.forename: Ai-Sheng
authors.initials: AS
authors.lastname: Xiong
mesh_terms: D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D056726:Genetic Association Studies; D031050:Oenanthe / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012965:Sodium Chloride / Q000494:pharmacology; D012984:Software; D013312:Stress, Physiological / Q000187:drug effects / Q000235:genetics*
chemical_list: D010940:Plant Proteins; D012965:Sodium Chloride