Pirrò Stefano, Zanella Letizia, Kenzo Maurice, Montesano Carla, Minutolo Antonella, Potestà Marina, Sobze Martin Sanou, Canini Antonella, Cirilli Marco, Muleo Rosario, Colizzi Vittorio, Galgani Andrea
Department of Biology, University of Rome "Tor Vergata", Rome, Italy.
Mir-Nat s.r.l., Rome, Italy.
PLoS One. 2016 Mar 1;11(3):e0149495. doi: 10.1371/journal.pone.0149495. eCollection 2016.
Moringa oleifera is a widespread plant with substantial nutritional and medicinal value. We postulated that microRNAs (miRNAs), which are endogenous, noncoding small RNAs regulating gene expression at the post-transcriptional level, might contribute to the medicinal properties of plants of this species after ingestion into human body, regulating human gene expression. However, the knowledge is scarce about miRNA in Moringa. Furthermore, in order to test the hypothesis on the pharmacological potential properties of miRNA, we conducted a high-throughput sequencing analysis using the Illumina platform. A total of 31,290,964 raw reads were produced from a library of small RNA isolated from M. oleifera seeds. We identified 94 conserved and two novel miRNAs that were validated by qRT-PCR assays. Results from qRT-PCR trials conducted on the expression of 20 Moringa miRNA showed that are conserved across multiple plant species as determined by their detection in tissue of other common crop plants. In silico analyses predicted target genes for the conserved miRNA that in turn allowed to relate the miRNAs to the regulation of physiological processes. Some of the predicted plant miRNAs have functional homology to their mammalian counterparts and regulated human genes when they were transfected into cell lines. To our knowledge, this is the first report of discovering M. oleifera miRNAs based on high-throughput sequencing and bioinformatics analysis and we provided new insight into a potential cross-species control of human gene expression. The widespread cultivation and consumption of M. oleifera, for nutritional and medicinal purposes, brings humans into close contact with products and extracts of this plant species. The potential for miRNA transfer should be evaluated as one possible mechanism of action to account for beneficial properties of this valuable species.
辣木是一种广泛分布的植物，具有很高的营养和药用价值。我们推测，微小RNA（miRNA）作为内源性非编码小RNA，在转录后水平调节基因表达，摄入人体后可能有助于该物种植物的药用特性，调节人类基因表达。然而，关于辣木中miRNA的知识却很少。此外，为了验证关于miRNA药理潜在特性的假设，我们使用Illumina平台进行了高通量测序分析。从辣木种子分离的小RNA文库中总共产生了31,290,964条原始读数。我们鉴定出94个保守的和2个新的miRNA，它们通过qRT-PCR分析得到验证。对20个辣木miRNA表达进行的qRT-PCR试验结果表明，这些miRNA在多种植物物种中保守，这是通过在其他常见作物植物组织中的检测确定的。计算机分析预测了保守miRNA的靶基因，进而使miRNA与生理过程的调节相关联。一些预测的植物miRNA与其哺乳动物对应物具有功能同源性，当它们转染到细胞系中时可调节人类基因。据我们所知，这是基于高通量测序和生物信息学分析发现辣木miRNA的首次报道，我们为人类基因表达的潜在跨物种调控提供了新的见解。辣木出于营养和药用目的而广泛种植和消费，使人类与该植物物种的产品和提取物密切接触。应评估miRNA转移的可能性，将其作为解释这种有价值物种有益特性的一种可能作用机制。
