Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
ID Marton, Kristina (Author), ID Flajšman, Marko (Author), ID Radišek, Sebastjan (Author), ID Košmelj, Katarina (Author), ID Jakše, Jernej (Author), ID Javornik, Branka (Author), ID Berne, Sabina (Author)

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The vascular plant pathogen Verticillium nonalfalfae causes Verticillium wilt in several important crops. VnaSSP4.2 was recently discovered as a V. nonalfalfae virulence effector protein in the xylem sap of infected hop. Here, we expanded our search for candidate secreted effector proteins (CSEPs) in the V. nonalfalfae predicted secretome using a bioinformatic pipeline built on V. nonalfalfae genome data, RNA-Seq and proteomic studies of the interaction with hop. The secretome, rich in carbohydrate active enzymes, proteases, redox proteins and proteins involved in secondary metabolism, cellular processing and signaling, includes 263 CSEPs. Several homologs of known fungal effectors (LysM, NLPs, Hce2, Cerato-platanins, Cyanovirin-N lectins, hydrophobins and CFEM domain containing proteins) and avirulence determinants in the PHI database (Avr-Pita1 and MgSM1) were found. The majority of CSEPs were non-annotated and were narrowed down to 44 top priority candidates based on their likelihood of being effectors. These were examined by spatio-temporal gene expression profiling of infected hop. Among the highest in planta expressed CSEPs, five deletion mutants were tested in pathogenicity assays. A deletion mutant of VnaUn.279, a lethal pathotype specific gene with sequence similarity to SAM-dependent methyltransferase (LaeA), had lower infectivity and showed highly reduced virulence, but no changes in morphology, fungal growth or conidiation were observed. Several putative secreted effector proteins that probably contribute to V. nonalfalfae colonization of hop were identified in this study. Among them, LaeA gene homolog was found to act as a potential novel virulence effector of V. nonalfalfae. The combined results will serve for future characterization of V. nonalfalfae effectors, which will advance our understanding of Verticillium wilt disease.

Keywords:plant fungal pathogens, protein domains, Verticillium, fungal genomics, fungal genetics, proteases, plant pathogens, gene expression
Work type:Article
Typology:1.01 - Original Scientific Article
Organization:BF - Biotechnical Faculty
Publication status:Published
Publication version:Version of Record
Number of pages:28 str.
Numbering:Vol. 13, iss. 6, art. e0198971
PID:20.500.12556/RUL-128210 This link opens in a new window
ISSN on article:1932-6203
DOI:10.1371/journal.pone.0198971 This link opens in a new window
COBISS.SI-ID:8940665 This link opens in a new window
Publication date in RUL:06.07.2021
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Record is a part of a journal

Title:PloS one
Publisher:Public Library of Science
COBISS.SI-ID:2005896 This link opens in a new window


License:CC BY 4.0, Creative Commons Attribution 4.0 International
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:12.06.2018

Secondary language

Keywords:Verticillium nonalfalfae, hmelj, genetika


Funder:ARRS - Slovenian Research Agency
Project number:P4-0077
Name:Kmetijske rastline - genetika in sodobne tehnologije

Funder:ARRS - Slovenian Research Agency
Project number:J4-8220
Name:Funkcionalna in strukturna analiza verticilijskih efektorskih proteinov in njihovih tarč v hmelju in modelnih rastlinah

Funder:ARRS - Slovenian Research Agency
Project number:342250

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