Flax rust infection transcriptomics reveals a transcriptional profile that may be indicative for rust Avr genes

PLoS One. 2019 Dec 12;14(12):e0226106. doi: 10.1371/journal.pone.0226106. eCollection 2019.

Abstract

Secreted effectors of fungal pathogens are essential elements for disease development. However, lack of sequence conservation among identified effectors has long been a problem for predicting effector complements in fungi. Here we have explored the expression characteristics of avirulence (Avr) genes and candidate effectors of the flax rust fungus, Melampsora lini. We performed transcriptome sequencing and real-time quantitative PCR (qPCR) on RNA extracted from ungerminated spores, germinated spores, isolated haustoria and flax seedlings inoculated with M. lini isolate CH5 during plant infection. Genes encoding two categories of M. lini proteins, namely Avr proteins and plant cell wall degrading enzymes (CWDEs), were investigated in detail. Analysis of the expression profiles of 623 genes encoding predicted secreted proteins in the M. lini transcriptome shows that the six known Avr genes (i.e. AvrM (avrM), AvrM14, AvrL2, AvrL567, AvrP123 (AvrP) and AvrP4) fall within a group of 64 similarly expressed genes that are induced in planta and show a peak of expression early in infection with a subsequent decline towards sporulation. Other genes within this group include two paralogues of AvrL2, an AvrL567 virulence allele, and a number of genes encoding putative effector proteins. By contrast, M. lini genes encoding CWDEs fall into different expression clusters with their distribution often unrelated to their catalytic activity or substrate targets. These results suggest that synthesis of M. lini Avr proteins may be regulated in a coordinated fashion and that the expression profiling-based analysis has significant predictive power for the identification of candidate Avr genes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Basidiomycota / genetics*
  • Basidiomycota / pathogenicity*
  • Computational Biology
  • Flax / genetics*
  • Flax / microbiology*
  • Fungal Proteins / genetics
  • Gene Expression Profiling
  • Gene Expression Regulation, Fungal
  • Genes, Fungal
  • Host-Pathogen Interactions / genetics*
  • Mycoses / genetics
  • Mycoses / microbiology
  • Plant Diseases / genetics
  • Plant Diseases / microbiology*
  • Plant Leaves / microbiology
  • Spores, Fungal / genetics
  • Transcriptome / physiology
  • Virulence / genetics
  • Virulence Factors / genetics*

Substances

  • Fungal Proteins
  • Virulence Factors

Grants and funding

This work was conducted with the support of the Australian Research Council grants DP1093850 (Role of fungal secreted proteins as plant disease effectors. ARH, DAJ, and PND) https://www.arc.gov.au/, DP130104098 (Molecular basis of rust infection and host plant resistance: ARH, DAJ, and PND) https://www.arc.gov.au/, and the China Scholarship Council grant (No.2010630010: WW) https://www.csc.edu.cn. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. KWS SAAT SE provided support for this study in the form of salary for first author AN. The commercial company: KWS SAAT SE did not have any role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of all authors are articulated in the “author contributions” section.