National Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Genetic Improvement of Soybean (General, Ministry of Agriculture), National Center for Soybean Improvement, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.
Subtropical Horticulture Research Institute, Jeju National University, Jeju, 63243, South Korea.
BMC Genomics. 2022 Mar 2;23(1):171. doi: 10.1186/s12864-022-08385-z.
Soybean mosaic virus (SMV) is one of the most devastating pathogens of soybean. MicroRNAs (miRNAs) are a class of non-coding RNAs (21-24 nucleotides) which are endogenously produced by the plant host as part of a general gene expression regulatory mechanisms, but also play roles in regulating plant defense against pathogens. However, miRNA-mediated plant response to SMV in soybean is not as well documented.
In this study, we analyzed 18 miRNA libraries, including three biological replicates from two soybean lines (Resistant and susceptible lines to SMV strain SC3 selected from the near-isogenic lines of Qihuang No. 1 × Nannong1138-2) after virus infection at three different time intervals (0 dpi, 7 dpi and 14 dpi). A total of 1,092 miRNAs, including 608 known miRNAs and 484 novel miRNAs were detected. Differential expression analyses identified the miRNAs profile changes during soybean-SMV interaction. Then, miRNAs potential target genes were predicted via data mining, and functional annotation was done by Gene Ontology (GO) analysis. The expression patterns of several miRNAs were validated by quantitative real-time PCR. We also validated the miRNA-target gene interaction by agrobacterium-mediated transient expression in Nicotiana benthamiana.
We have identified a large number of miRNAs and their target genes and also functional annotations. We found that multiple miRNAs were differentially expressed in the two lines and targeted a series of NBS-LRR resistance genes. It is worth mentioning that many of these genes exist in the previous fine-mapping interval of the resistance gene locus. Our study provides additional information on soybean miRNAs and an insight into the role of miRNAs during SMV-infection in soybean.
大豆花叶病毒(SMV)是大豆最具破坏性的病原体之一。MicroRNAs(miRNAs)是一类非编码 RNA(21-24 个核苷酸),它们作为植物宿主中普遍基因表达调控机制的一部分内源性产生,但也在调节植物对病原体的防御中发挥作用。然而,miRNA 介导的大豆对 SMV 的反应尚未得到很好的记录。
在这项研究中,我们分析了 18 个 miRNA 文库,包括来自两个大豆品系(从 Qihuang No.1×Nannong1138-2 的近等基因系中选择的对 SMV 菌株 SC3 具有抗性和敏感性的品系)在三个不同时间间隔(0dpi、7dpi 和 14dpi)病毒感染后的三个生物学重复。总共检测到 1092 个 miRNA,包括 608 个已知 miRNA 和 484 个新 miRNA。差异表达分析确定了大豆-SMV 相互作用过程中 miRNA 谱的变化。然后,通过数据挖掘预测了 miRNA 潜在靶基因,并通过基因本体论(GO)分析进行了功能注释。通过定量实时 PCR 验证了几个 miRNA 的表达模式。我们还通过农杆菌介导的瞬时表达在 Nicotiana benthamiana 中验证了 miRNA-靶基因的相互作用。
我们已经鉴定了大量的 miRNA 及其靶基因和功能注释。我们发现,两个品系中多个 miRNA 表达差异,靶向一系列 NBS-LRR 抗性基因。值得一提的是,这些基因中的许多存在于抗性基因座的先前精细定位区间内。我们的研究为大豆 miRNA 提供了更多信息,并深入了解了 miRNA 在大豆 SMV 感染过程中的作用。