He Xiaohong, Sun Quan, Jiang Huaizhong, Zhu Xiaoyan, Mo Jianchuan, Long Lu, Xiang Liuxin, Xie Yongfang, Shi Yuzhen, Yuan Youlu, Cai Yingfan
College of Bioinformation, Chongqing University of Posts and Telecommunications, Chongqing, 400065 China.
State Key Laboratory of Cotton Biology, Henan Key Laboratory of Plant Stress Biology, College of Life Sciences, Henan University, Kaifeng, 475004 China.
Springerplus. 2014 Sep 27;3:564. doi: 10.1186/2193-1801-3-564. eCollection 2014.
Plant microRNAs (miRNAs) play essential roles in the post-transcriptional regulation of gene expression during development, flowering, plant growth, metabolism, and stress responses. Verticillium wilt is one of the vascular disease in plants, which is caused by the Verticillium dahlia and leads to yellowing, wilting, lodging, damage to the vascular tissue, and death in cotton plants. Upland cotton varieties KV-1 have shown resistance to Verticillium wilt in multiple levels. However, the knowledge regarding the post-transcriptional regulation of the resistance is limited. Here two novel small RNA (sRNA) libraries were constructed from the seedlings of upland cotton variety KV-1, which is highly resistant to Verticillium wilts and inoculated with the V991 and D07038 Verticillium dahliae (V. dahliae) of different virulence strains. Thirty-seven novel miRNAs were identified after sequencing these two libraries by the Illumina Solexa system. According to sequence homology analysis, potential target genes of these miRNAs were predicted. With no more than three sequence mismatches between the novel miRNAs and the potential target mRNAs, we predicted 49 target mRNAs for 24 of the novel miRNAs. These target mRNAs corresponded to genes were found to be involved in plant-pathogen interactions, endocytosis, the mitogen-activated protein kinase (MAPK) signaling pathway, and the biosynthesis of isoquinoline alkaloid, terpenoid backbone, primary bile acid and secondary metabolites. Our results showed that some of these miRNAs and their relative gene are involved in resistance to Verticillium wilts. The identification and characterization of miRNAs from upland cotton could help further studies on the miRNA regulatory mechanisms of resistance to Verticillium wilt.
植物微小RNA（miRNA）在植物发育、开花、生长、代谢及应激反应过程中的基因表达转录后调控中发挥着重要作用。黄萎病是植物的一种维管束病害，由大丽轮枝菌引起，可导致棉花植株发黄、枯萎、倒伏、维管束组织受损及死亡。陆地棉品种KV-1在多个水平上表现出对黄萎病的抗性。然而，关于这种抗性的转录后调控的认识还很有限。在此，从对黄萎病具有高度抗性的陆地棉品种KV-1的幼苗中构建了两个新型小RNA（sRNA）文库，这些幼苗接种了不同毒力菌株的V991和D07038大丽轮枝菌（V. dahliae）。通过Illumina Solexa系统对这两个文库进行测序后，鉴定出了37个新型miRNA。根据序列同源性分析，预测了这些miRNA的潜在靶基因。在新型miRNA与潜在靶mRNA之间的序列错配不超过三个的情况下，我们为24个新型miRNA预测了49个靶mRNA。发现这些与靶mRNA对应的基因参与植物-病原体相互作用、内吞作用、丝裂原活化蛋白激酶（MAPK）信号通路以及异喹啉生物碱、萜类骨架、初级胆汁酸和次生代谢物的生物合成。我们的结果表明，其中一些miRNA及其相关基因参与了对黄萎病的抗性。从陆地棉中鉴定和表征miRNA有助于进一步研究miRNA对黄萎病抗性的调控机制。
