Designer transcription activator-like effectors enable discovery of cell death-inducer genes

verfasst von

Roxana A Roeschlin, Sepideh M Azad, René P Grove, Ana Chuan, Lucila García, Regina Niñoles, Facundo Uviedo, Liara Villalobos-Piña, Maria E Massimino, María R Marano, Jens Boch, José Gadea

Abstract

TALEs (transcription activator-like effectors) in plant-pathogenic Xanthomonas bacteria activate expression of plant genes and support infection or cause a resistance response. PthA4AT is a TALE with a particularly short DNA-binding domain harbouring only 7.5-repeats which triggers cell death in Nicotiana benthamiana; however, the genetic basis for this remains unknown. To identify possible target genes of PthA4AT that mediate cell death in N. benthamiana, we exploited the modularity of TALEs to stepwise enhance their specificity and reduce potential target sites. Substitutions of individual repeats suggested that PthA4AT-dependent cell death is sequence-specific. Stepwise addition of repeats to the C-terminal or N-terminal end of the repeat region narrowed the sequence requirements in promoters of target genes. Transcriptome profiling and in silico target prediction allowed the isolation of two cell death-inducer genes, which encode a patatin-like protein and a bifunctional monodehydroascorbate reductase/carbonic anhydrase protein. These two proteins are not linked to known TALE-dependent resistance genes. Our results show that the aberrant expression of different endogenous plant genes can cause a cell death reaction, which supports the hypothesis that TALE-dependent executor resistance genes can originate from various plant processes. Our strategy further demonstrates the use of TALEs to scan genomes for genes triggering cell death and other relevant phenotypes.

Organisationseinheit(en)

Institut für Pflanzengenetik

Externe Organisation(en)

Instituto Nacional de Tecnología Agropecuaria (INTA)
Instituto de Biología Molecular y Celular de Rosario (IBR)-Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET)
Universidad Politecnica de Valencia
Universidad Nacional de Rosario

Typ

Artikel

Journal

Plant Physiology

Band

195

Seiten

2985–2996

Anzahl der Seiten

12

ISSN

0032-0889

Publikationsdatum

08.2024

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Genetik, Physiologie, Pflanzenkunde

Elektronische Version(en)

https://doi.org/10.1093/plphys/kiae230 (Zugang: Offen)