Genome-wide profiling of sRNAs in the Verticillium dahliae-infected Arabidopsis roots.
Genome-wide profiling of sRNAs in the Verticillium dahliae-infected Arabidopsis roots.
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受大丽黄萎病菌感染的拟南芥根部 sRNA 的全基因组分析
DOI：
10.1080/21501203.2018.1426062
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Zhao JH
中科院分区：
文献类型：
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作者：
Jin Y;Zhao P;Fang YY;Gao F;Guo HS;Zhao JH
小RNA （sRNAs，包括小干扰RNA [sirna]和微RNA [miRNAs]）是RNA沉默（或RNA干扰）的关键介质，在植物发育和对生物和非生物刺激的反应中起着重要作用。黄萎病（Verticillium wilt）是由大丽花黄萎病（Verticillium dahliae）等土传真菌病原菌引起的植物血管病。我们之前报道了大丽花v感染增加了两个植物内源性mirna，这些mirna输出到真菌细胞以沉默毒力基因。为了研究植物对大丽花侵染的全基因组反应，本研究分别从感染和未感染大丽花的拟南芥根中构建了两个sRNA文库。通过sRNA测序，共发现31个保守mirna在大丽花感染早期存在差异表达。其中，miR160、miR164、miR166、miR167、miR390和miR156h的表达量通过northern blot检测。逆转录定量实时聚合酶链反应结果显示，在大丽花感染后诱导mirna （miR160、miR164、miR166和miR167）分别下调其靶基因（ARF10、NAC1、PHV和ARF6）的表达。此外，我们确定了从两个文库的不同区域产生的特定的阶段性sirna。这些miRNAs和sRNAs的分析为进一步了解和利用宿主诱导的基因沉默策略来控制植物维管病原体奠定了基础。
ABSTRACT Small RNAs (sRNAs, including small interfering RNAs [siRNAs] and micro RNAs [miRNAs]) are key mediators of RNA silencing (or RNA interference), which play important roles in plant development and response to biotic and abiotic stimulation. Verticillium wilt is a plant vascular disease caused by the soil-borne fungal pathogens, such as Verticillium dahliae. We previously reported that V. dahliae infection increased two plant endogenous miRNAs that were exported to fungal cell to silence virulence genes. To investigate plant sRNAs in genome-wide response to V. dahliae infection, in this study, we constructed two sRNA libraries from Arabidopsis roots with and without V. dahliae infection, respectively. In total, 31 conserved miRNAs were found to be differentially expressed during the early stage of infection with V. dahliae using sRNA sequencing. Among these, the expression levels of miR160, miR164, miR166, miR167, miR390 and miR156h were confirmed by northern blot. Reverse transcription quantitative real time polymerase chain reaction results showed that the induction of miRNAs (miR160, miR164, miR166 and miR167) upon V. dahliae infection downregulated the expression of their targeted genes (ARF10, NAC1, PHV and ARF6), respectively. In addition, we identified specific phased siRNAs generated from distinct regions of two libraries. Profiling of these miRNAs and sRNAs lay the foundation for further understanding and utilising the host-induced gene silencing strategy to control plant vascular pathogens.