State Key Laboratory of Grassland Agro-ecosystems, Key Laboratory of Grassland Livestock Industry Innovation, Ministry of Agriculture and Rural Affairs, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, 730020, P.R. China.
Guangxi Institute of Animal Sciences, Nanning, 530001, P.R. China.
Sci Rep. 2018 Sep 20;8(1):14138. doi: 10.1038/s41598-018-32153-3.
MicroRNAs (miRNAs) exhibit diverse and important roles in regulation of various biological processes at the post-transcriptional level in plants. In this study, Melilotus albus miRNA and their target genes were elucidated from five M. albus near-isogenic lines which differ in coumarin content to construct small RNA libraries through high-throughput sequencing. A total of 417 known miRNAs and 76 novel miRNAs were identified in M. albus. In addition, 4155 different target genes for 114 known miRNA families and 14 target genes for 2 novel miRNAs were identified in M. albus. Moreover, mtr-miR5248 and mtr-miR7701-5p target c35498_g3 and gma-miR396a-3p target c37211_g1 involved in coumarin biosynthesis were identified by using the differential expression of the miRNAs and their target genes correlation analysis. The abundance of miRNAs and potential target genes were validated by qRT-PCR analysis. We also found that there were both positive and negative expression changing patterns between miRNAs and their related target genes. Our first and preliminary study of miRNAs will contribute to our understanding of the functions and molecular regulatory mechanisms of miRNAs and their target genes, and provide information on regulating the complex coumarin pathway in M. albus for future research.
MicroRNAs (miRNAs) 在植物的转录后水平上表现出多样化和重要的调节作用。在这项研究中,从五种香豆素含量不同的白三叶草近等基因系中阐明了 Melilotus albus miRNA 及其靶基因,通过高通量测序构建了小 RNA 文库。在白三叶草中鉴定出了 417 个已知 miRNA 和 76 个新的 miRNA。此外,在白三叶草中鉴定出了 114 个已知 miRNA 家族的 4155 个不同靶基因和 2 个新 miRNA 的 14 个靶基因。此外,通过 miRNA 及其靶基因相关性分析,鉴定出了参与香豆素生物合成的 mtr-miR5248 和 mtr-miR7701-5p 靶标 c35498_g3 和 gma-miR396a-3p 靶标 c37211_g1。通过 qRT-PCR 分析验证了 miRNA 和潜在靶基因的丰度。我们还发现 miRNA 和相关靶基因之间存在着正表达和负表达的变化模式。我们对 miRNA 的首次初步研究将有助于我们理解 miRNA 和其靶基因的功能和分子调控机制,并为未来研究调节白三叶草中复杂的香豆素途径提供信息。