Validation of Suitable Reference Genes for Assessing Gene Expression of MicroRNAs in Lonicera japonica.
Validation of Suitable Reference Genes for Assessing Gene Expression of MicroRNAs in Lonicera japonica.
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用于评估忍冬中 MicroRNA 基因表达的合适参考基因的验证
DOI：
10.3389/fpls.2016.01101
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发表时间：
2016
影响因子：
5.6
通讯作者：
Huang L
中科院分区：
文献类型：
--
作者：
Wang Y;Liu J;Wang X;Liu S;Wang G;Zhou J;Yuan Y;Chen T;Jiang C;Zha L;Huang L
MicroRNA（miRNAs）在植物次生代谢和环境响应中起着重要的调控作用，可作为中草药不同品种和产地的生物标志物。然而，有限的信息是从日本，这是广泛使用的东亚国家，由于各种药物活性的次级代谢产物的miRNAs。选择合适的参考基因，通过实时荧光定量PCR（qRT）定量检测目的miRNA的表达，对于阐明不同组织和不同品种乳杆菌次级代谢调控的分子机制具有重要意义。日本。为了精确标准化L.在日本，16个候选miRNAs在三个组织中，以及从16个生产区域收集的21个栽培品种中，使用GeNorm，Normands和Referral算法进行了检查。我们的结果显示u534122和u3868172的组合是所有样品中最好的参考基因。通过测定不同品种的循环阈值（Ct）范围，证实了它们的特异性。从不同产区收集的日本粳稻中，这表明使用这两种参考miRNA足以用于不同组织、品种和产区的准确转录物标准化。据我们所知，这是第一个报告验证参考miRNAs在金银花（忍冬属）。本研究的结果可以进一步促进发现不同品种L.日本。
MicroRNAs (miRNAs), which play crucial regulatory roles in plant secondary metabolism and responses to the environment, could be developed as promising biomarkers for different varieties and production areas of herbal medicines. However, limited information is available for miRNAs from Lonicera japonica, which is widely used in East Asian countries owing to various pharmaceutically active secondary metabolites. Selection of suitable reference genes for quantification of target miRNA expression through quantitative real-time (qRT)-PCR is important for elucidating the molecular mechanisms of secondary metabolic regulation in different tissues and varieties of L. japonica. For precise normalization of gene expression data in L. japonica, 16 candidate miRNAs were examined in three tissues, as well as 21 cultivated varieties collected from 16 production areas, using GeNorm, NormFinder, and RefFinder algorithms. Our results revealed combination of u534122 and u3868172 as the best reference genes across all samples. Their specificity was confirmed by detecting the cycling threshold (Ct) value ranges in different varieties of L. japonica collected from diverse production areas, suggesting the use of these two reference miRNAs is sufficient for accurate transcript normalization with different tissues, varieties, and production areas. To our knowledge, this is the first report on validation of reference miRNAs in honeysuckle (Lonicera spp.). Restuls from this study can further facilitate discovery of functional regulatory miRNAs in different varieties of L. japonica.