title: Validation of reference genes for quantitative real-time PCR during latex regeneration in rubber tree.
abstract: In rubber tree, latex regeneration is one of the decisive factors influencing the rubber yield, although its molecular regulation is not well known. Quantitative real-time PCR (qPCR) is a popular and powerful tool used to understand the molecular mechanisms of latex regeneration. However, the suitable reference genes required for qPCR are not available to investigate the expressions of target genes during latex regeneration. In this study, 20 candidate reference genes were selected and evaluated for their expression stability across the samples during the process of latex regeneration. All reference genes showed a relatively wide range of the threshold cycle values, and their stability was validated by four different algorithms (comparative delta Ct method, Bestkeeper, NormFinder and GeNorm). Three softwares (comparative delta Ct method, NormFinder and GeNorm) exported similar results that identify UBC4, ADF, UBC2a, eIF2 and ADF4 as the top five suitable references, and 18S as the least suitable one. The application of the screened references would improve accuracy and reliability of gene expression analysis in latex regeneration experiments.
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China.
authors: Xiangyu
authors: X
authors: Long
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China; College of Agronomy, Hainan University, Haikou, Hainan 570228, PR China.
authors: Bin
authors: B
authors: He
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China.
authors: Xinsheng
authors: X
authors: Gao
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China.
authors: Yunxia
authors: Y
authors: Qin
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China.
authors: Jianghua
authors: J
authors: Yang
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China.
authors: Yongjun
authors: Y
authors: Fang
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China.
authors: Jiyan
authors: J
authors: Qi
authors: Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, PR China. Electronic address: chaorongtang@126.com.
authors: Chaorong
authors: C
authors: Tang
mesh_terms: D000465:Algorithms; D020869:Gene Expression Profiling / Q000379:methods; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics*; D028482:Hevea / Q000235:genetics* / Q000378:metabolism; D007840:Latex / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012015:Reference Standards; D012038:Regeneration / Q000235:genetics*; D015203:Reproducibility of Results
chemical_list: D007840:Latex
keywords: Algorithms; Hevea brasiliensis; Latex regeneration; Quantitative real-time PCR; Reference genes