Cell-specific RNA profiling reveals host genes expressed in Arabidopsis cells haustoriated by downy mildew

Shuta Asai, Volkan Cevik, Jonathan D G Jones, Ken Shirasu

Research output: Contribution to journalArticlepeer-review

2 Citations (SciVal)

Abstract

The downy mildew oomycete Hyaloperonospora arabidopsidis, an obligate filamentous pathogen, infects Arabidopsis (Arabidopsis thaliana) by forming structures called haustoria inside host cells. Previous transcriptome analyses have revealed that host genes are specifically induced during infection; however, RNA profiling from whole-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 translating ribosome affinity purification system using 2 high-affinity binding proteins, colicin E9 and Im9 (immunity protein of colicin E9), applicable to 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 insights into the Arabidopsis–downy mildew interaction. We propose that our protocol for profiling cell-specific transcripts will apply to several stimulus-specific contexts and other plant–pathogen interactions.
Original languageEnglish
Article numberkiad326
Number of pages12
JournalPlant Physiology
DOIs
Publication statusPublished - 12 Jun 2023

Bibliographical note

Funding:
This work was supported by JSPS KAKENHI Grant Numbers 17K07679 (S.A.), 20H02995 (S.A.), JP20H05909 (K.S.), and JP22H00364 (K.S.) and by funding to TSL from the Gatsby Foundation (J.D.G.J).

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