Jeyaraj Anburaj, Wang Xuewen, Wang Shuangshuang, Liu Shengrui, Zhang Ran, Wu Ailin, Wei Chaoling
State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei, China.
Department of Biotechnology, Karpagam Academy of Higher Education, Coimbatore, India.
Front Plant Sci. 2019 Sep 12;10:1096. doi: 10.3389/fpls.2019.01096. eCollection 2019.
Anthracnose disease is caused by , and is common in leaves of the tea plant (). MicroRNAs (miRNAs) have been known as key modulators of gene expression in response to environmental stresses, disease resistance, defense responses, and plant immunity. However, the role of miRNAs in responses to remains unexplored in tea plant. Therefore, in the present study, six miRNA sequencing data sets and two degradome data sets were generated from -inoculated and control tea leaves. A total of 485 conserved and 761 novel miRNAs were identified. Of those, 239 known and 369 novel miRNAs exhibited significantly differential expression under stress. One thousand one hundred thirty-four and 596 mRNAs were identified as targets of 389 conserved and 299 novel miRNAs by degradome analysis, respectively. Based on degradome analysis, most of the predicted targets are negatively correlated with their corresponding conserved and novel miRNAs. The expression levels of 12 miRNAs and their targets were validated by quantitative real-time PCR. A negative correlation between expression profiles of five miRNAs (PC-5p-80764_22, csn-miR160c, csn-miR828a, csn-miR164a, and csn-miR169e) and their targets (WRKY, ARF, MYB75, NAC, and NFY transcription factor) was observed. The predicted targets of five interesting miRNAs were further validated through 5'RLM-RACE. Furthermore, Gene Ontology and metabolism pathway analysis revealed that most of the target genes were involved in the regulation of auxin pathway, ROS scavenging pathway, salicylic acid mediated pathway, receptor kinases, and transcription factors for plant growth and development as well as stress responses in tea plant against stress. This study enriches the resources of stress-responsive miRNAs and their targets in and thus provides novel insights into the miRNA-mediated regulatory mechanisms, which could contribute to the enhanced susceptibility of in tea plant.
炭疽病由[病原体名称未给出]引起,在茶树([茶树学名未给出])叶片中很常见。微小RNA(miRNA)是已知的响应环境胁迫、抗病性、防御反应和植物免疫过程中基因表达的关键调节因子。然而,miRNA在茶树对[病原体名称未给出]反应中的作用尚未得到探索。因此,在本研究中,从接种[病原体名称未给出]和对照的茶叶中生成了六个miRNA测序数据集和两个降解组数据集。共鉴定出485个保守miRNA和761个新miRNA。其中,239个已知miRNA和369个新miRNA在[病原体名称未给出]胁迫下表现出显著差异表达。通过降解组分析,分别有1134个和596个mRNA被鉴定为389个保守miRNA和299个新miRNA的靶标。基于降解组分析,大多数预测靶标与其相应的保守和新miRNA呈负相关。通过定量实时PCR验证了12个miRNA及其靶标的表达水平。观察到五个miRNA(PC-5p-80764_22、csn-miR160c、csn-miR828a、csn-miR164a和csn-miR169e)与其靶标(WRKY、ARF、MYB75、NAC和NFY转录因子)的表达谱之间呈负相关。通过5'RLM-RACE进一步验证了五个有趣miRNA的预测靶标。此外,基因本体论和代谢途径分析表明,大多数靶基因参与生长素途径、活性氧清除途径、水杨酸介导途径、受体激酶以及茶树生长发育和对[病原体名称未给出]胁迫的应激反应中的转录因子的调控。本研究丰富了茶树中胁迫响应miRNA及其靶标的资源,从而为miRNA介导的调控机制提供了新的见解,这可能有助于提高茶树对[病原体名称未给出]的易感性。