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Archive > Volume 15 Issue 1 > 2023,15(1):69-75. DOI:https://doi.org/10.1016/j.chmed.2022.08.006 Prev Next

人參microRNA的鑒定及靶基因分析

Original articles

Identification of microRNA and analysis of target genes in Panax ginseng

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    摘要:
    目的：人參中的主要有效成分人參皂苷、多糖和酚類物質(zhì)在3~5年生的園參和十年以上的石柱參中的含量差異不顯著，主要化合物不能完全解釋它們之間功效上的差異。根據(jù)金銀花(Lonicerae Japonicae Flos)和甘草(Glycyrrhizae Radix et Rhizoma)的報(bào)道，microRNA可能在藥效中發(fā)揮作用，因此我們?cè)诓煌L(zhǎng)年限的人參中鑒定了microRNA，并分析了它們的靶基因。 方法：采用高通量測(cè)序技術(shù)，構(gòu)建了人參RNA-Seq、small RNA-Seq和降解基因組數(shù)據(jù)庫(kù)。通過(guò)qRT-PCR鑒定差異表達(dá)的microRNAs。 結(jié)果：從人參根部共獲得63,875個(gè)unigenes和24,154,579個(gè)small RNA clean reads。通過(guò)生物信息學(xué)靶標(biāo)預(yù)測(cè)軟件從這些small RNA中識(shí)別出71個(gè)miRNA家族，包括34個(gè)保守的miRNAs，37個(gè)非保守的miRNA家族，以及17個(gè)已知miRNAs的179個(gè)靶基因。通過(guò)降解組測(cè)序和計(jì)算，我們最終驗(yàn)證了參與轉(zhuǎn)錄、能量代謝、生物脅迫和抗病的8個(gè)miRNAs中的13個(gè)靶點(diǎn)，表明miRNAs在人參的發(fā)育中具有重要意義。一直以來(lái)，主要的miRNA靶標(biāo)表現(xiàn)出組織特異性和表達(dá)模式的復(fù)雜性。 結(jié)論：不同生長(zhǎng)年限的人參(石柱參和園參)存在差異表達(dá)的microRNAs，miRNA靶標(biāo)在人參中的調(diào)控作用和功能注釋有待進(jìn)一步研究。

    Abstract:
    Objective: Ginsenosides, polysaccharides and phenols, the main active ingredients in Panax ginseng, are not different significantly in content between 3 and 5 years old of ginsengs called Yuan ginseng and more than ten years old ones called Shizhu ginseng. The responsible chemical compounds cannot fully explain difference in efficacy between them. According to reports in Lonicerae Japonicae Flos (Jinyinhua in Chinese) and Glycyrrhizae Radix et Rhizoma (Gancao in Chinese), microRNA may play a role in efficacy, so we identified microRNAs in P. ginseng at the different growth years and analyzed their target genes. Methods: Using high-throughput sequencing, the RNA-Seq, small RNA-Seq and degradome databases of P. ginseng were constructed. The differentially expressed microRNAs was identified by qRT-PCR. Results: A total of 63,875 unigenes and 24,154,579 small RNA clean reads were obtained from the roots of P. ginseng. From these small RNAs, 71 miRNA families were identified by bioinformatics target prediction software, including 34 conserved miRNAs, 37 non-conserved miRNA families, as well as 179 target genes of 17 known miRNAs. Through degradome sequencing and computation, we finally verified 13 targets of eight miRNAs involved in transcription, energy metabolism, biological stress and disease resistance, suggesting the significance of miRNAs in the development of P. ginseng. Consistently, major miRNA targets exhibited tissue specificity and complexity in expression patterns. Conclusion: Differential expression microRNAs were found in different growth years of ginsengs (Shizhu ginseng and Yuan ginseng), and the regulatory roles and functional annotations of miRNA targets in P. ginseng need further investigation.

    關(guān)鍵詞:
    高通量測(cè)序;miRNA;靶基因;人參

    Keywords:


    Project Supported:
    This work was supported by the National Natural Science Foundation for Young Scholars of China (No. 81403195).

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