Agromonas: a rapid disease assay for Pseudomonas syringae growth in agroinfiltrated leaves

Buscaill, Pierre, Sanguankiattichai, Nattapong, Lee, Yoon Joo, Kourelis, Jiorgos ORCID: https://orcid.org/0000-0002-9007-1333, Preston, Gail and van der Hoorn, Renier A. L. (2021) Agromonas: a rapid disease assay for Pseudomonas syringae growth in agroinfiltrated leaves. Plant Journal, 105 (3). pp. 831-840. ISSN 0960-7412

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Abstract

The lengthy process to generate transformed plants is a limitation in current research on the interactions of the model plant pathogen Pseudomonas syringae with plant hosts. Here we present an easy method called agromonas, where we quantify P. syringae growth in agroinfiltrated leaves of Nicotiana benthamiana using a cocktail of antibiotics to select P. syringae on plates. As a proof of concept, we demonstrate that transient expression of PAMP receptors reduces bacterial growth, and that transient depletion of a host immune gene and transient expression of a type-III effector increase P. syringae growth in agromonas assays. We show that we can rapidly achieve structure−function analysis of immune components and test the function of immune hydrolases. The agromonas method is easy, fast and robust for routine disease assays with various Pseudomonas strains without transforming plants or bacteria. The agromonas assay offers a reliable approach for further comprehensive analysis of plant immunity.

Item Type: Article
Additional Information: Acknowledgements: The authors thank Alan Collmer for providing the DC3000() mutant; Yuki Ichinose for providing the 6605 strain; Tolga Bozkurt for providing the pRNAiGG vector; Nicola J. Patron and Sylvestre Marillonnet for providing Golden Gate plasmids; Ursula Pyzio for plant care; Sarah Rodgers and Caroline O’Brien for technical support. ERC Consolidator grant 616447 ‘GreenProteases’ (PB, RH); BBSRC grants BB/R017913/1 (PB, RH); the Royal Thai Government Scholarship (NS); the Clarendon foundation (JK); the Oxford Interdisciplinary Bioscience DTP (BB/M011224/1 to NS, GP).
Uncontrolled Keywords: agrobacterium,disease assay,nicotiana benthamiana,plant immunity,pseudomonas syringae,technical advance,genetics,plant science,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1311
Faculty \ School: Faculty of Science > The Sainsbury Laboratory
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Depositing User: LivePure Connector
Date Deposited: 07 Dec 2023 01:46
Last Modified: 01 Feb 2024 03:07
URI: https://ueaeprints.uea.ac.uk/id/eprint/93896
DOI: 10.1111/tpj.15056

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