Nitrate transporter protein NPF5.12 and major latex-like protein MLP6 are important defense factors against Verticillium longisporum

Abstract

Plant defense responses to the soil-borne fungus Verticillium longisporum causing stem stripe disease on oilseed rape (Brassica napus) are poorly understood. In this study, a population of recombinant inbred lines (RILs) using the Arabidopsis accessions Sei-0 and Can-0 was established. Composite interval mapping, transcriptome data, and T-DNA mutant screening identified the NITRATE/PEPTIDE TRANSPORTER FAMILY 5.12 (AtNPF5.12) gene as being associated with disease susceptibility in Can-0. Co-immunoprecipitation revealed interaction between AtNPF5.12 and the MAJOR LATEX PROTEIN family member AtMLP6, and fluorescence microscopy confirmed this interaction in the plasma membrane and endoplasmic reticulum. CRISPR/Cas9 technology was applied to mutate the NPF5.12 and MLP6 genes in B. napus. Elevated fungal growth in the npf5.12 mlp6 double mutant of both oilseed rape and Arabidopsis demonstrated the importance of these genes in defense against V. longisporum. Colonization of this fungus depends also on available nitrates in the host root. Accordingly, the negative effect of nitrate depletion on fungal growth was less pronounced in Atnpf5.12 plants with impaired nitrate transport. In addition, suberin staining revealed involvement of the NPF5.12 and MLP6 genes in suberin barrier formation. Together, these results demonstrate a dependency on multiple plant factors that leads to successful V. longisporum root infection.Recognition of the fungal pathogen Verticillium longisporum triggers nitrate starvation and MLP6-mediated defenses in Arabidopsis, together reducing the suberin barrier.

Keywords

Arabidopsis; Brassica napus; major latex protein; MLP6; nitrate; NPF5.12; Verticillium longisporum

Published in

Journal of Experimental Botany
2024, volume: 75, number: 13, pages: 4148-4164
Publisher: OXFORD UNIV PRESS

SLU Authors

• Dölfors, Fredrik

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

• Roos, Jonas

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

• Bejai, Sarosh

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

• Fogelqvist, Johan

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

• Dixelius, Christina

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Botany

Publication identifier

• DOI: https://doi.org/10.1093/jxb/erae185

Permanent link to this page (URI)

https://res.slu.se/id/publ/130841