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A vector system for fast-forward in vivo studies of the ZAR1 resistosome in the model plant Nicotiana benthamiana

View ORCID ProfileAdeline Harant, View ORCID ProfileToshiyuki Sakai, View ORCID ProfileSophien Kamoun, View ORCID ProfileHiroaki Adachi
doi: https://doi.org/10.1101/2020.05.15.097584
Adeline Harant
The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, Norwich NR4 7UH, UK
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Toshiyuki Sakai
The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, Norwich NR4 7UH, UK
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Sophien Kamoun
The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, Norwich NR4 7UH, UK
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  • For correspondence: sophien.kamoun@tsl.ac.uk hiroaki.adachi@tsl.ac.uk
Hiroaki Adachi
The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, Norwich NR4 7UH, UK
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  • For correspondence: sophien.kamoun@tsl.ac.uk hiroaki.adachi@tsl.ac.uk
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ABSTRACT

Nicotiana benthamiana has emerged as a complementary experimental system to Arabidopsis. It enables fast-forward in vivo analyses primarily through transient gene expression and is particularly popular in the study of plant immunity. Recently, our understanding of NLR plant immune receptors has greatly advanced following the discovery of Arabidopsis ZAR1 resistosome. Here, we describe a novel vector system of 52 plasmids that enables functional studies of the ZAR1 resistosome in N. benthamiana. We showed that ZAR1 stands out among the coiled coil class of NLRs for being highly conserved across distantly related dicot plant species and confirmed NbZAR1 as the N. benthamiana ortholog of Arabidopsis ZAR1. NbZAR1 triggers autoimmune cell death in N. benthamiana and this activity is dependent on a functional N-terminal α1 helix. C-terminally tagged NbZAR1 remains functional in N. benthamiana thus enabling cell biology and biochemical studies in this plant system. We conclude that the NbZAR1 open source plasmids form an additional experimental system to Arabidopsis for in planta resistosome studies.

Competing Interest Statement

S.K. receives funding from industry on NLR biology.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted May 16, 2020.
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A vector system for fast-forward in vivo studies of the ZAR1 resistosome in the model plant Nicotiana benthamiana
Adeline Harant, Toshiyuki Sakai, Sophien Kamoun, Hiroaki Adachi
bioRxiv 2020.05.15.097584; doi: https://doi.org/10.1101/2020.05.15.097584
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A vector system for fast-forward in vivo studies of the ZAR1 resistosome in the model plant Nicotiana benthamiana
Adeline Harant, Toshiyuki Sakai, Sophien Kamoun, Hiroaki Adachi
bioRxiv 2020.05.15.097584; doi: https://doi.org/10.1101/2020.05.15.097584

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