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Identification of novel microRNAs in the Verticillium wilt-resistant upland cotton variety KV-1 by high-throughput sequencing

机译:高通量测序鉴定耐黄萎病陆地棉品种KV-1中的新型microRNA

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摘要

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.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-3-564) contains supplementary material, which is available to authorized users.
机译:植物微RNA(miRNA)在发育,开花,植物生长,代谢和胁迫反应期间,在基因表达的转录后调控中发挥重要作用。黄萎病是植物中的一种血管疾病,由黄萎病菌引起,导致黄化,枯萎,倒伏,对血管组织的损害以及棉花植物的死亡。陆地棉品种KV-1对多株黄萎病表现出抗性。但是,关于抗性的转录后调控的知识是有限的。在这里,从陆地棉品种KV-1的幼苗中构建了两个新颖的小RNA(sRNA)库,该品种对黄萎病具有高度抗性,并分别接种了不同毒力菌株的V991和D07038大麦黄萎病菌(V. dahliae)。通过Illumina Solexa系统对这两个文库进行测序后,鉴定出了37种新颖的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与潜在靶标mRNA之间的序列错配不超过三个的情况下,我们预测了24种新型miRNA的49种靶标mRNA。这些靶标mRNA对应于与植物-病原体相互作用,内吞作用,有丝分裂原有关的基因。激活的蛋白激酶(MAPK)信号通路,以及异喹啉生物碱,萜类骨架,初级胆汁酸和次级代谢产物的生物合成。我们的结果表明,其中一些miRNA及其相关基因与黄萎病的抗性有关。陆地棉中miRNA的鉴定和表征可以帮助进一步研究黄萎病抗性的miRNA调控机制。电子补充材料本文的在线版本(doi:10.1186 / 2193-1801-3-564)包含补充材料,其中适用于授权用户。

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