Selection of suitable inner reference genes for normalisation of microRNA expression response to abiotic stresses by RT-qPCR in leaves, flowers and young stems of peach
Selection of suitable inner reference genes for normalisation of microRNA expression response to abiotic stresses by RT-qPCR in leaves, flowers and young stems of peach
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DOI：
10.1016/j.scienta.2013.10.030
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发表时间：
2014-01-22
影响因子：
4.3
通讯作者：
Gao, Zhihong
中科院分区：
文献类型：
--
作者：
Luo, Xiaoyan;Shi, Ting;Gao, Zhihong
RT-qPCR是目前研究基因表达最准确、最灵敏、应用最广泛的技术。然而，选择合适的内参基因是获得准确结果的关键因素。到目前为止，已有大量研究检测到miRNA在不同实验条件下的表达，为其生物学功能的研究提供了便利，而对参考基因在植物中的研究很少，在桃树(Prunus Persica L)中还没有。因此，本研究旨在寻找适合桃miRNA表达正常化的参考基因(S)。本研究以2个已报道的看家基因和15个PPE-miRNAs(Prunus Persica MiRNAs)为参考候选基因，研究了它们在桃不同组织以及干旱、盐分和内涝胁迫处理下的表达稳定性。经过分析和验证，miR5059和miR5072两对基因是最好的参考基因，这可能有助于进一步研究miRNA在桃非生物胁迫和其他发育过程中的功能。(C)2013爱思唯尔B.V.保留所有权利。
RT-qPCR is currently the most accurate, sensitive and widely used technique to investigate gene expression. However, the selection of a suitable internal reference gene is a crucial factor for an accurate and precise result. To date, numerous studies have detected miRNA expression in different experimental conditions to facilitate studies of their biological function, whereas only few studies on reference genes have been performed in plants and none in peach (Prunus persica L). Therefore, the present study aimed to identify suitable reference gene(s) for the normalisation of miRNA expression in peach. In this study, two previously reported housekeeping genes and 15 ppe-miRNAs (Prunus persica miRNAs) were selected as reference candidates and their expression stability was investigated in different peach tissues as well as following drought stress, salinity stress, and waterlogging stress treatments. After analysis and validation, the pair of miR5059 and miR5072 genes was found to be the best reference genes, which might aid further studies on miRNA function in response to abiotic stresses and other development processes in peach. (C) 2013 Elsevier B.V. All rights reserved.