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高通量测序揭示了 fenoxaprop-P-ethyl 抗性稗草中 miRNA 的差异调控。

High-throughput sequencing reveals differential regulation of miRNAs in fenoxaprop-P-ethyl-resistant Beckmannia syzigachne.

机构信息

College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.

Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education, Nanjing 210095, China.

出版信息

Sci Rep. 2016 Jun 29;6:28725. doi: 10.1038/srep28725.

Abstract

Non-target site resistance (NTSR) to herbicides is an increasing concern for weed control. The majority of previous studies have focused on metabolic resistance mechanisms of NTSR, but no research exists on gene regulation mechanisms behind herbicide resistance, such as microRNA (miRNA). Here, we identified 3 American sloughgrass (Beckmannia syzigachne Steud.) populations containing fenoxaprop-P-ethyl-resistant plants. We then constructed small RNA libraries and subjected them to deep sequencing and bioinformatics analyses. Forty known and 36 potentially novel, predicted miRNAs were successfully identified. Of these, we identified 3 conserved, predicted candidate NTSR-determinant miRNAs and their potential corresponding target genes, as well as 4 novel potential miRNAs with high count. Target gene prediction and annotation indicated that these 7 differentially expressed miRNAs potentially play a role in regulating specific stress-responsive genes, very likely related to herbicide resistance. Expression profiles were determined with quantitative real-time PCR. The present study is a novel, large-scale characterization of weed miRNAs. The results should further our understanding of miRNA expression profiles associated with herbicide resistance, allowing for the development of more effective weed management strategies.

摘要

非靶标位点抗除草剂性(NTSR)是杂草防治中越来越令人关注的问题。以前的大多数研究都集中在 NTSR 的代谢抗性机制上,但对于除草剂抗性背后的基因调控机制,如 microRNA(miRNA),则没有研究。在这里,我们鉴定了 3 个含有精噁唑禾草灵抗性植株的美国野黍(Beckmannia syzigachne Steud.)种群。然后,我们构建了小 RNA 文库,并进行了深度测序和生物信息学分析。成功鉴定出 40 个已知和 36 个潜在的新预测 miRNA。其中,我们鉴定出 3 个保守的、预测的候选 NTSR 决定 miRNA 及其潜在的对应靶基因,以及 4 个具有高计数的新型潜在 miRNA。靶基因预测和注释表明,这 7 个差异表达的 miRNA 可能在调节特定的应激响应基因中发挥作用,很可能与除草剂抗性有关。采用定量实时 PCR 确定了表达谱。本研究是对杂草 miRNA 的一项新颖的、大规模的描述。研究结果将进一步加深我们对与除草剂抗性相关的 miRNA 表达谱的理解,从而为制定更有效的杂草管理策略提供依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/063e/4926119/088eef271715/srep28725-f1.jpg

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