Inducible TRAP RNA profiling reveals host genes expressed in Arabidopsis cells haustoriated by downy mildew
Inducible TRAP RNA profiling reveals host genes expressed in Arabidopsis cells haustoriated by downy mildew
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DOI：
10.1101/2023.01.17.524393
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发表时间：
2023-01
期刊：
影响因子：
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通讯作者：
S. Asai;V. Cevik;Jonathan D. G. Jones;K. Shirasu
中科院分区：
文献类型：
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作者：
S. Asai;V. Cevik;Jonathan D. G. Jones;K. Shirasu
霜霉病卵菌透明霜霉是一种专性丝状病原体，通过在宿主细胞内形成称为吸器的取食结构来感染拟南芥。先前的转录组分析显示，宿主基因在感染期间被特异性诱导;然而，来自感染组织的RNA谱分析可能无法捕获仅发生在吸器化宿主细胞中的关键转录事件，其中病原体注射毒力效应子以调节宿主免疫力。为了研究拟南芥与H.在细胞水平上，我们设计了一种新的翻译核糖体亲和纯化系统，适用于诱导型启动子，包括病原体响应性启动子，从而使吸器化细胞特异性RNA谱。在H.在拟南芥吸器化细胞中，我们发现了促进对病原体的易感性或抗性的基因，为拟南芥/霜霉病相互作用提供了新的见解。我们建议，我们的新协议，分析细胞特异性转录将适用于几个刺激特定的背景和其他植物病原体的相互作用。
The downy mildew oomycete Hyaloperonospora arabidopsidis, an obligate filamentous pathogen, infects Arabidopsis by forming feeding structures called haustoria inside host cells. Previous transcriptome analyses revealed host genes are specifically induced during infection; however, RNA profiling from infected tissues may fail to capture key transcriptional events occurring exclusively in haustoriated host cells where the pathogen injects virulence effectors to modulate host immunity. To determine interactions between Arabidopsis and H. arabidopsidis at the cellular level, we devised a new translating ribosome affinity purification system applicable to inducible, including pathogen-responsive, promoters thus enabling haustoriated cell-specific RNA profiling. Among the host genes specifically expressed in H. arabidopsidis-haustoriated cells, we found genes that promote either susceptibility or resistance to the pathogen, providing new insights into the Arabidopsis/downy mildew interaction. We propose that our novel protocol for profiling cell-specific transcripts will be applicable to several stimulus-specific contexts and other plant-pathogen interactions.