Instituto de Biotecnología, CICVyA, INTA Castelar, CP 1686 Hurlingham, Buenos Aires, Argentina.
BMC Plant Biol. 2009 Dec 30;9:152. doi: 10.1186/1471-2229-9-152.
Micro RNAs (miRs) constitute a large group of endogenous small RNAs that have crucial roles in many important plant functions. Virus infection and transgenic expression of viral proteins alter accumulation and activity of miRs and so far, most of the published evidence involves post-transcriptional regulations.
Using transgenic plants expressing a reporter gene under the promoter region of a characterized miR (P-miR164a), we monitored the reporter gene expression in different tissues and during Arabidopsis development. Strong expression was detected in both vascular tissues and hydathodes. P-miR164a activity was developmentally regulated in plants with a maximum expression at stages 1.12 to 5.1 (according to Boyes, 2001) along the transition from vegetative to reproductive growth. Upon quantification of P-miR164a-derived GUS activity after Tobacco mosaic virus Cg or Oilseed rape mosaic virus (ORMV) infection and after hormone treatments, we demonstrated that ORMV and gibberellic acid elevated P-miR164a activity. Accordingly, total mature miR164, precursor of miR164a and CUC1 mRNA (a miR164 target) levels increased after virus infection and interestingly the most severe virus (ORMV) produced the strongest promoter induction.
This work shows for the first time that the alteration of miR pathways produced by viral infections possesses a transcriptional component. In addition, the degree of miR alteration correlates with virus severity since a more severe virus produces a stronger P-miR164a induction.
Micro RNAs(miRs)是一组重要的内源性小分子 RNA,在许多重要的植物功能中发挥着关键作用。病毒感染和病毒蛋白的转基因表达改变了 miRs 的积累和活性,到目前为止,大多数已发表的证据涉及转录后调控。
我们使用表达报告基因的转基因植物,该报告基因受已鉴定的 miR(P-miR164a)启动子区域的调控,在不同组织和拟南芥发育过程中监测报告基因的表达。在维管束组织和水孔中均检测到强烈的表达。在从营养生长向生殖生长过渡阶段 1.12 到 5.1(根据 Boyes,2001),P-miR164a 的活性受到发育调控。在用烟草花叶病毒 Cg 或油菜花叶病毒(ORMV)感染以及激素处理后,对 P-miR164a 衍生的 GUS 活性进行定量后,我们证明 ORMV 和赤霉素提高了 P-miR164a 的活性。相应地,总成熟 miR164、miR164a 的前体和 CUC1 mRNA(miR164 的靶标)水平在病毒感染后增加,有趣的是,最严重的病毒(ORMV)产生了最强的启动子诱导。
这项工作首次表明,病毒感染引起的 miR 途径的改变具有转录成分。此外,miR 改变的程度与病毒的严重程度相关,因为更严重的病毒会产生更强的 P-miR164a 诱导。