DNA methylation plays a role on in vitro culture induced loss of virulence in Botrytis cinerea

BioRxiv : the Preprint Server for Biology
Jimmy BreenCarlos Marcelino Rodriguez Lopez


Little is known about the mechanisms causing loss of virulence in pathogenic fungi as a result of protracted culture. We studied the extent to which patterns of DNA methylation varied between virulent and reduced virulence derivative cultures of Botrytis cinerea, and identify the genes/genomic regions affected by these epigenetic modifications. B. cinerea was cultured in vitro for eight months involving subculture every four weeks. Fungal conidia were harvested at every four-week subculturing stage and inoculated onto Arabidopsis thaliana Col-0 plants for virulence testing. Global epi/genetic changes in B. cinerea during culture were assessed using methylation-sensitive amplified polymorphisms (MSAPs) on mycelium from eight different sub-culture time points and from mycelium recovered after eight months in culture and then inoculated onto A. thaliana. Culture induced epi/allele characterisation was carried out by whole genome sequencing and bisulfite sequencing of gDNA from samples after two and eight months in culture and after 8 months in culture and following inoculation onto an A. thaliana plant. Virulence declined with time in culture and recovered after one fungal generation on A. thaliana. MSAP data show that epi/genetic...Continue Reading

Related Concepts

Gene Polymorphism
Hydrogen sulfite
DNA Methylation [PE]
Baptisia cinerea
Botrytis cinerea
Protein Methylation
Regulation of Biological Process
Pathogenic Organism

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