Phylogenetic Diversity and Phenotypic Characterization of Phlyctema vagabunda (syn. Neofabraea alba) and Neofabraea kienholzii Causing Postharvest Bull's Eye Rot of Apple in Northern Italy

Bull's eye rot, caused by Phlyctema vagabunda and Neofabraea species, is one of the most important postharvest diseases of apple. South Tyrol (northern Italy) is the largest continuous apple-producing area in Europe, with approximately 1 million tons being produced yearly and conserved in technologically advanced storage facilities for several months. Still, studies on the pathogen species causing postharvest bull's eye rot of apple, as well as their diversity and biology, are lacking for this region. Therefore, the main purpose of the present work was to identify and characterize fungal isolates obtained from decayed apple fruit with symptoms of bull's eye rot that were collected in 2018 and 2019 in different packinghouses in South Tyrol. Among more than 1,000 fungal isolates that were obtained, 419 could be assigned to the genera Phlyctema and/or Neofabraea based on rot symptoms on apple fruit and colony morphology on potato dextrose agar. A smaller subset of 101 representative isolates was further analyzed by DNA sequencing of the internal transcribed spacer region. Furthermore, partial segments of the beta-tubulin gene, the translation elongation factor 1 alpha gene, and the 16S mitochondrial ribosomal RNA gene were studied. The phylogenetic analyses, including sequences of reference species, showed that P. vagabunda is the dominant species associated with bull's eye rot of apple in the study area, whereas Neofabraea kienholzii was found only on a small number of apple fruit samples. The combination of multilocus sequence data revealed 11 unique genotypes that belonged to P. vagabunda and four to N. kienholzii. To the best of our knowledge, this study is the first to report N. kienholzii as a postharvest pathogen of apple in Italy. Finally, a pathogenicity test demonstrated different degrees of virulence among selected isolates of P. vagabunda and N. kienholzii on the cultivar Golden Delicious. The present study emphasizes the importance of accurate species identification, because different species may vary in their biological and pathogenic characteristics, and consequently require distinct disease management strategies, both in the field and during the postharvest stages.

Automated summary

Bull's eye rot, caused by Phlyctema vagabunda and Neofabraea species, is a significant postharvest disease of apples. This study investigated the diversity and biology of these pathogens in South Tyrol, Italy, which is the largest apple-producing region in Europe. The dominant species associated with bull's eye rot in this area was identified as P. vagabunda, while N. kienholzii was found only in a small number of samples. Genetic analysis revealed the presence of unique genotypes within these species. Pathogenicity tests demonstrated variations in virulence among different isolates. Accurate species identification is crucial for effective disease management.