Robust transcriptional indicators of immune cell death revealed by spatiotemporal transcriptome analyses
Salguero Linares, Jose (Centre de Recerca en Agrigenòmica)
Serrano, Irene (Université de Toulouse)
Ruiz Solaní, Nerea (Centre de Recerca en Agrigenòmica)
Salas Gómez, Marta (Centre de Recerca en Agrigenòmica)
Phukan, Ujjal Jyoti (Centre de Recerca en Agrigenòmica)
González, Víctor Manuel (Centre de Recerca en Agrigenòmica)
Bernardo-Faura, Martí (Centre de Recerca en Agrigenòmica)
Valls, Marc (Centre de Recerca en Agrigenòmica)
Rengel, David (Université de Toulouse)
Sánchez Coll, Núria (Centre de Recerca en Agrigenòmica)

Fecha:	2022
Resumen:	Recognition of a pathogen by the plant immune system often triggers a form of regulated cell death traditionally known as the hypersensitive response (HR). This type of cell death occurs precisely at the site of pathogen recognition, and it is restricted to a few cells. Extensive research has shed light on how plant immune receptors are mechanistically activated. However, two central key questions remain largely unresolved: how does cell death zonation take place, and what are the mechanisms that underpin this phenomenon? Consequently, bona fide transcriptional indicators of HR are lacking, which prevents deeper insight into its mechanisms before cell death becomes macroscopic and precludes early or live observation. In this study, to identify the transcriptional indicators of HR we used the paradigmatic Arabidopsis thaliana-Pseudomonas syringae pathosystem and performed a spatiotemporally resolved gene expression analysis that compared infected cells that will undergo HR upon pathogen recognition with bystander cells that will stay alive and activate immunity. Our data revealed unique and time-dependent differences in the repertoire of differentially expressed genes, expression profiles, and biological processes derived from tissue undergoing HR and that of its surroundings. Furthermore, we generated a pipeline based on concatenated pairwise comparisons between time, zone, and treatment that enabled us to define 13 robust transcriptional HR markers. Among these genes, the promoter of an uncharacterized AAA-ATPase was used to obtain a fluorescent reporter transgenic line that displays a strong spatiotemporally resolved signal specifically in cells that will later undergo pathogen-triggered cell death. This valuable set of genes can be used to define cells that are destined to die upon infection with HR-triggering bacteria, opening new avenues for specific and/or high-throughput techniques to study HR processes at a single-cell level.
Ayudas:	Agencia Estatal de Investigación PID2019-108595RB-I00 Agencia Estatal de Investigación AGL2016-78002-R Agencia Estatal de Investigación PID2019-108595RB-I00 Agencia Estatal de Investigación BES-2017-080210 Agencia Estatal de Investigación FPU19/03778 Ministerio de Economía y Competitividad SEV-2015-0533 Ministerio de Ciencia e Innovación CEX2019-000902-S European Commission 267196
Nota:	Altres ajuts: CERCA Programme/Generalitat de Catalunya
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió acceptada per publicar
Materia:	Arabidopsis thaliana ; Cell death indicator ; Effector-triggered immunity ; Hypersensitive response ; Pattern-triggered immunity ; Plant immunity ; Pseudomonas syringae
Publicado en:	Molecular Plant, Vol. 15, Issue 6 (June 2022) , p. 1059-1075

DOI: 10.1016/j.molp.2022.04.010

Postprint 105 p, 9.0 MB

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