Yin Fuqiang, Gao Jian, Liu Ming, Qin Cheng, Zhang Wenyou, Yang Aiguo, Xia Mingzhong, Zhang Zhiming, Shen Yaou, Lin Haijian, Luo Chenggang, Pan Guangtang
Maize Research Institute of Sichuan Agricultural University/Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region, Ministry of Agriculture, Chengdu, 611130, China.
Funct Integr Genomics. 2014 Jun;14(2):319-32. doi: 10.1007/s10142-014-0365-4. Epub 2014 Mar 25.
MicroRNAs (miRNAs) play a pivotal role in post-transcriptional regulation of gene expression in plants. In this study, we investigate miRNAs in an agronomically important common tobacco in China, named Honghua Dajinyuan (a drought-tolerant cultivar). Here, we report a comprehensive analysis of miRNA expression profiles in mock-treat grown (CK) and 20 % polyethylene glycol-grown (PEG-grown) tobacco roots using a high-throughput sequencing approach. A total of 656 unique miRNAs representing 53 miRNA families were identified in the two libraries, of which 286 unique miRNAs representing 162 microRNAs were differentially expressed. In addition, nine differentially expressed microRNAs selected from different expressed miRNA family with high abundance were subjected to further analysis and validated by quantitative real-time PCR (Q-PCR). In addition, the expression pattern of these identified candidate conserved miRNA and target genes of three identified miRNA (nta-miR172b, nta-miR156i, and nta-miR160a) were also validated by Q-PCR. Gene ontology (GO) enrichment analysis suggests that the putative target genes of these differentially expressed miRNAs are involved in metabolic process and response to stimulus. In particular, 25 target genes are involved in regulating plant hormone signal transduction and metabolism, indicating that these association microRNAs may play important regulatory roles in responding to PEG resistance. Moreover, this study adds a significant number of novel miRNAs to the tobacco miRNome.
微小RNA（miRNA）在植物基因表达的转录后调控中起关键作用。在本研究中，我们对中国一种具有重要农艺价值的普通烟草——红花大金元（一个耐旱品种）中的miRNA进行了研究。在此，我们报告了使用高通量测序方法对模拟处理生长（CK）和20%聚乙二醇处理生长（PEG处理）的烟草根中miRNA表达谱的综合分析。在两个文库中总共鉴定出代表53个miRNA家族的656个独特miRNA，其中代表162个微小RNA的286个独特miRNA差异表达。此外，从不同表达的高丰度miRNA家族中选择的9个差异表达微小RNA进行了进一步分析，并通过定量实时PCR（Q-PCR）进行了验证。此外，还通过Q-PCR验证了这些鉴定出的候选保守miRNA以及三个鉴定出的miRNA（nta-miR172b、nta-miR156i和nta-miR160a）的靶基因的表达模式。基因本体（GO）富集分析表明，这些差异表达miRNA的推定靶基因参与代谢过程和对刺激的反应。特别是，25个靶基因参与调节植物激素信号转导和代谢，表明这些相关miRNA可能在响应PEG抗性中发挥重要调节作用。此外，本研究为烟草miRNome增加了大量新的miRNA。
