LmCBP1, a secreted chitin-binding protein, is required for the pathogenicity of Leptosphaeria maculans on Brassica napus

Fungal Genet Biol. 2020 Mar;136:103320. doi: 10.1016/j.fgb.2019.103320. Epub 2019 Dec 19.

Abstract

Leptosphaeria maculans is the causal agent of blackleg disease on Brassica napus. Determining the underlying functions of genes required for pathogenesis is essential for understanding the infection process. A chitin-binding protein (LmCBP1) was discovered as a pathogenicity factor for the infection of B. napus by L. maculans through gene knockout using the CRISPR-Cas9 system. Chitin-binding activity was demonstrated through a chitin-protein binding assay. A secreted signal peptide was detected using a yeast secreted-signal peptide trap assay. An increased expression level during the infection stage was also observed, suggesting that LmCBP1 is a secreted protein. The knockout mutants showed decreased infection on B. napus, with reduced pathogenicity on ten cultivars with/without diverse R genes. The mutants were more sensitive to H2O2 compared to wild type L. maculans isolate JN3. This study provides evidence of the virulence of a novel chitin-binding protein LmCBP1 on B. napus through mutants created via the CRISPR-Cas9 system.

Keywords: Blackleg; Brassica napus; CRISPR/Cas9; Canola; Chitin binding protein; Hydrogen peroxide; Leptosphaeria maculans; LmCBP1; Pathogenicity; ROS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brassica napus / microbiology*
  • CRISPR-Cas Systems
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism
  • Chitin / metabolism
  • DNA, Fungal
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism
  • Gene Knockout Techniques
  • Host-Pathogen Interactions
  • Hydrogen Peroxide / pharmacology
  • Leptosphaeria / genetics*
  • Leptosphaeria / metabolism
  • Leptosphaeria / pathogenicity*
  • Phylogeny
  • Plant Diseases / microbiology*
  • Reactive Oxygen Species / metabolism
  • Virulence / genetics

Substances

  • Carrier Proteins
  • DNA, Fungal
  • Fungal Proteins
  • Reactive Oxygen Species
  • Chitin
  • Hydrogen Peroxide

Supplementary concepts

  • Leptosphaeria maculans