Validation of reference genes for quantitative real-time PCR during latex regeneration in rubber tree
Validation of reference genes for quantitative real-time PCR during latex regeneration in rubber tree
复制标题
橡胶树乳胶再生过程中定量实时 PCR 参考基因的验证
DOI：
10.1016/j.gene.2015.03.026
复制
发表时间：
2015-06-01
期刊：
影响因子：
3.5
通讯作者：
Tang, Chaorong
中科院分区：
文献类型：
--
作者：
Long, Xiangyu;He, Bin;Tang, Chaorong
在橡胶树中，胶乳再生是影响橡胶产量的决定性因素之一，但其分子调控尚不清楚。实时定量聚合酶链式反应(QPCR)是研究胶乳再生分子机制的有力工具。然而，没有合适的qPCR所需的参考基因来研究目的基因在胶乳再生过程中的表达。在这项研究中，选择了20个候选参考基因，并评估了它们在胶乳再生过程中跨样本表达的稳定性。所有参考基因的阈值周期范围都比较宽，并用四种不同的算法(比较增量Ct法、BestKeeper法、NormFinder法和GeNorm法)验证了它们的稳定性。3个软件(比较增量CT法、NormFinder和GeNorm)输出了相似的结果，确定UBC4、ADF、UBC2a、eIF2和ADF4是最合适的5个参考文献，18S是最不合适的参考文献。筛选的参考文献的应用将提高胶乳再生实验中基因表达分析的准确性和可靠性。(C)2015爱思唯尔B.V.保留所有权利。
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. (C) 2015 Elsevier B.V. All rights reserved.