Analysis of the laccase gene family and miR397-/miR408-mediated posttranscriptional regulation in Salvia miltiorrhiza
Analysis of the laccase gene family and miR397-/miR408-mediated posttranscriptional regulation in Salvia miltiorrhiza
复制标题
丹参漆酶基因家族及miR397/miR408介导的转录后调控分析
DOI：
10.7717/peerj.7605
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发表时间：
2019-08-29
期刊：
影响因子：
2.7
通讯作者：
Lu, Shanfa
中科院分区：
文献类型：
--
作者：
Li, Caili;Li, Dongqiao;Lu, Shanfa
丹参是最常用的中药材之一。它含有重要的生物活性酚类化合物，如丹酚酸、类黄酮和花青素。阐明酚类化合物的生物合成及其调控机制对于丹参品质改良具有重要意义。漆酶（LAC）是一种含多铜的酶，可能参与酚类化合物的聚合。迄今为止，人们对丹参中的 LAC 基因知之甚少。通过对丹参全基因组序列和转录组的系统研究，我们鉴定了65个全长SmLAC基因（SmLAC1-SmLAC65）。系统发育分析表明，62 个已鉴定的 SmLAC 与来自拟南芥和毛果杨的 LAC 聚类在 7 个进化枝（C1-C7）中，而其他 3 个属于丹参特异性进化枝（C8）。所有推导的SmLAC蛋白都含有4个保守的特征序列和3个典型的Cu氧化酶结构域，并且来自丹参、拟南芥和毛果的大多数LAC的基因结构高度保守，然而编码C8蛋白的SmLAC表现出独特的内含子-外显子结构。这表明植物LAC在基因结构上具有保守性和多样性。大多数SmLACs表现出组织特异性表达模式，表明SmLACs在丹参中发挥多种功能。高通量小RNA序列和降解组数据的分析以及使用5'RACE方法的实验验证表明，23个SmLAC是Smi-miR397的靶标。其中，三个也是 Smi-miR408 的目标。这表明 miR397 和 miR408 在 SmLAC 基因转录后调控中的重要性。我们的结果为进一步证明SmLACs在丹参生产生物活性酚类化合物中的功能奠定了基础。
Salvia miltiorrhiza is one of the most commonly used traditional Chinese medicine materials. It contains important bioactive phenolic compounds, such as salvianolic acids, flavonoids and anthocyanins. Elucidation of phenolic compound biosynthesis and its regulatory mechanism is of great significance for S. miltiorrhiza quality improvement. Laccases (LACs) are multicopper-containing enzymes potentially involved in the polymerization of phenolic compounds. So far, little has been known about LAC genes in S. miltiorrhiza. Through systematic investigation of the whole genome sequence and transcriptomes of S. miltiorrhiza, we identified 65 full-length SmLAC genes (SmLAC1-SmLAC65). Phylogenetic analysis showed that 62 of the identified SmLACs clustered with LACs from Arabidopsis and Populus trichocarpa in seven clades (C1-C7), whereas the other three fell into one S. miltiorrhiza-specific clade (C8). All of the deduced SmLAC proteins contain four conserved signature sequences and three typical Cu-oxidase domains, and gene structures of most LACs from S. miltiorrhiza, Arabidopsis and P. trichocarpa were highly conserved, however SmLACs encoding C8 proteins showed distinct intron-exon structures. It suggests the conservation and diversity of plant LACs in gene structures. The majority of SmLACs exhibited tissue-specific expression patterns, indicates manifold functions of SmLACs played in S. miltiorrhiza. Analysis of high-throughput small RNA sequences and degradome data and experimental validation using the 5' RACE method showed that 23 SmLACs were targets of Smi-miR397. Among them, three were also targeted by Smi-miR408. It suggests the significance of miR397 and miR408 in posttranscriptional regulation of SmLAC genes. Our results provide a foundation for further demonstrating the functions of SmLACs in the production of bioactive phenolic compounds in S. miltiorrhiza.