title: Selection of suitable reference genes for qPCR normalization under abiotic stresses and hormone stimuli in carrot leaves.
abstract: Carrot, a biennial herb of the Apiaceae family, is among the most important vegetable crops in the world. In this study, nine candidate reference genes (GAPDH, ACTIN, eIF-4α, PP2A, SAND, TIP41, UBQ, EF-1α, and TUB) were cloned from carrot. Carrot plants were subjected to abiotic stresses (heat, cold, salt, and drought) and hormone stimuli (gibberellin, salicylic acid, methyl jasmonate, and abscisic acid). The expression profiles of the candidate reference genes were evaluated in three technical and biological replicates. Real-time qPCR data analyses were performed using three commonly used Excel-based applets namely, BestKeeper, geNorm, and NormFinder. ACTIN and TUB were the most stable genes identified among all sample groups, but individual analysis revealed changes in their expression profiles. GAPDH displayed the maximum stability for most of single stresses. To further validate the suitability of the reference genes identified in this study, the expression profile of DcDREB-A1 gene (homolog of AtDREB-A1 gene of Arabidophsis) was studied in carrot. The appropriate reference genes were selected that showed stable expression under the different experimental conditions.
authors: State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
authors: Chang
authors: C
authors: Tian
authors: State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
authors: Qian
authors: Q
authors: Jiang
authors: State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
authors: Feng
authors: F
authors: Wang
authors: State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
authors: Guang-Long
authors: GL
authors: Wang
authors: State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
authors: Zhi-Sheng
authors: ZS
authors: Xu
authors: State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.
authors: Ai-Sheng
authors: AS
authors: Xiong
mesh_terms: D000040:Abscisic Acid / Q000494:pharmacology; D000085:Acetates / Q000494:pharmacology; D003080:Cold Temperature; D003517:Cyclopentanes / Q000494:pharmacology; D018552:Daucus carota / Q000187:drug effects / Q000235:genetics*; D055864:Droughts; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000502:physiology; D020043:Genes, Essential; D017343:Genes, Plant; D005875:Gibberellins / Q000494:pharmacology; D006358:Hot Temperature; D054883:Oxylipins / Q000494:pharmacology; D010937:Plant Growth Regulators / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D020156:Salicylic Acid / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological / Q000235:genetics*
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D005875:Gibberellins; D054883:Oxylipins; D010937:Plant Growth Regulators; D012965:Sodium Chloride; D000040:Abscisic Acid; C072239:methyl jasmonate; D020156:Salicylic Acid