Quantitative Hormone Signaling Output Analyses of Arabidopsis thaliana Interactions With Virulent and Avirulent Hyaloperonospora arabidopsidis Isolates at Single-Cell Resolution

2020 | journal article. A publication with affiliation to the University of Göttingen.

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​Quantitative Hormone Signaling Output Analyses of Arabidopsis thaliana Interactions With Virulent and Avirulent Hyaloperonospora arabidopsidis Isolates at Single-Cell Resolution​
Ghareeb, H. ; El-Sayed, M.; Pound, M.; Tetyuk, O.; Hanika, K.; Herrfurth, C.   & Feussner, I.  et al.​ (2020) 
Frontiers in Plant Science11.​ DOI: https://doi.org/10.3389/fpls.2020.603693 

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Authors
Ghareeb, Hassan ; El-Sayed, Mohamed; Pound, Michael; Tetyuk, Olena; Hanika, Katharina; Herrfurth, Cornelia ; Feussner, Ivo ; Lipka, Volker 
Abstract
The phytohormones salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) are central regulators of biotic and abiotic stress responses in Arabidopsis thaliana. Here, we generated modular fluorescent protein-based reporter lines termed COLORFUL-PR1pro, -VSP2pro, and -PDF1.2apro. These feature hormone-controlled nucleus-targeted transcriptional output sensors and the simultaneous constitutive expression of spectrally separated nuclear reference and plasma membrane-localized reporters. This set-up allowed the study of cell-type specific hormone activities, cellular viability and microbial invasion. Moreover, we developed a software-supported high-throughput confocal microscopy imaging protocol for output quantification to resolve the spatio-temporal dynamics of respective hormonal signaling activities at single-cell resolution. Proof-of-principle analyses in A. thaliana leaves revealed distinguished hormone sensitivities in mesophyll, epidermal pavement and stomatal guard cells, suggesting cell type-specific regulatory protein activities. In plant-microbe interaction studies, we found that virulent and avirulent Hyaloperonospora arabidopsidis (Hpa) isolates exhibit different invasion dynamics and induce spatio-temporally distinct hormonal activity signatures. On the cellular level, these hormone-controlled reporter signatures demarcate the nascent sites of Hpa entry and progression, and highlight initiation, transduction and local containment of immune signals.
Issue Date
2020
Publisher
Frontiers Media S.A.
Journal
Frontiers in Plant Science 
eISSN
1664-462X
Language
English
Sponsor
Open-Access-Publikationsfonds 2020

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