Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China.
J Biosci. 2020;45.
miRNAs are important regulatory components involving in many biological processes, including plant development, vegetative and reproductive growth, and stress response. However, identification and characterization of miRNAs still remain limited for conifer species. In this study, with deep sequencing, we obtained 1,314,450 unique reads with 18-30 nt length from a stress-tolerant conifer, . We identified 37 conserved and 103 novel miRNAs, their unique characteristics were further analyzed, and 10 randomly selected were validated by qRT-PCR. Through miRNA target predictions and annotations, we found miRNA may have several targets as well a target could be regulated by several miRNAs, and a total of 2,397 mRNAs were predicted to be targets of the 140 miRNAs. These targets included not only important transcription factors such as auxin response factors, but also indispensable non-transcriptional factor proteins. Pathway-based analysis showed that miRNAs are involved in 172 metabolic pathways, of which 3 were discovered in adaptation-related pathways, indicating their possible relevance to the species' stress-tolerance characteristics. This study is expected to lay the foundation for exploring the regulative roles of miRNAs in development, growth, and response to environmental stresses of .
miRNAs 是重要的调控组成部分，参与许多生物过程，包括植物发育、营养和生殖生长以及应激反应。然而，对于针叶树物种，miRNAs 的鉴定和表征仍然有限。在这项研究中，通过深度测序，我们从一种耐应激的针叶树中获得了 1,314,450 条具有 18-30nt 长度的独特reads。我们鉴定了 37 个保守和 103 个新的 miRNAs，进一步分析了它们的独特特征，并通过 qRT-PCR 验证了 10 个随机选择的 miRNAs。通过 miRNA 靶标预测和注释，我们发现 miRNA 可能有几个靶标，一个靶标也可能被几个 miRNAs 调控，总共预测了 2,397 个 mRNAs 为 140 个 miRNAs 的靶标。这些靶标不仅包括生长素响应因子等重要的转录因子，还包括不可或缺的非转录因子蛋白。基于途径的分析表明，miRNAs 参与了 172 个代谢途径，其中 3 个在适应相关途径中被发现，表明它们可能与该物种的应激耐受特性有关。本研究有望为探索 miRNAs 在发育、生长和对环境应激的响应中的调控作用奠定基础。
