Botrytis cinerea loss and restoration of virulence during in vitro culture follows flux in global DNA methylation

James Breen, Luis Alejandro Jose Mur, Anushen Sivakumaran, Aderemi Akinyemi, Michael James Wilkinson, Carlos M. Rodriguez Lopez

Research output: Contribution to journalArticle


Pathogenic fungi can lose virulence after protracted periods of culture but little is known of the underlying mechanisms. Here we present the first single-base resolution methylome for the plant pathogen B. cinerea and identify differentially methylated genes/genomic regions associated with virulence erosion.

Cultures were maintained for eight months with subcultures and virulence testing every month. Methylation-sensitive amplified polymorphisms were performed at monthly intervals to characterise global changes to the pathogen’s genome during culture and also on DNA from mycelium inoculated onto Arabidopsis thaliana after eight months in culture. Characterisation of culture-induced epialleles was assessed by whole-genome re-sequencing and whole-genome bisulfite sequencing.

Virulence declined with time in culture and recovered after inoculation on A. thaliana. Variation detected by methylation-sensitive amplified polymorphisms followed virulence changes during culture. Whole-genome (bisulfite) sequencing showed marked changes on global and local methylation during culture but no significant genetic changes.

We imply that virulence is a non-essential plastic character that is at least partly modified by changing levels of DNA methylation during culture. We hypothesise that changing DNA methylation during culture may be responsible for the high virulence/low virulence transition in B. cinerea and speculate that this may offer fresh opportunities to control pathogen virulence.
Original languageEnglish
Pages (from-to)1-49
Number of pages49
Publication statusE-pub ahead of print - 26 Aug 2016

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