Diversity and Pathogenicity of Diaporthe Species Revealed from a Survey of Blueberry Orchards in Portugal

Blueberries (Vaccinium corymbosum) are widely cultivated worldwide and largely consumed due to their known antioxidant and medicinal properties. Although Diaporthe species have been documented in Portugal as causal agents of blueberry twig blight and dieback, there is still scarce information on the species that cause these symptoms. Moreover, Diaporthe vaccinii, recently synonymized with D. eres, has been considered a concern to blueberry production worldwide. However, the current knowledge about its impact on blueberries remains unclear. The diversity of Diaporthe species associated with diseased blueberry plants were assessed through a national survey. A multilocus sequence analysis of the rDNA internal transcribed spacer (ITS) region, the translation elongation factor 1-alpha (tef1-alpha), beta-tubulin (tub2), calmodulin (cal) and histone 3 (his3) genes unveiled the presence of Diaporthe ambigua, D. amygdali, D. crousii, D. foeniculina, D. hybrida, D. leucospermi, D. malorum and D. rudis. Moreover, all species were fully characterized based on a detailed morphological description. Diaporthe amygdali, D. hybrida, D. leucospermi and D. malorum are reported for the first time on diseased blueberries in Portugal. Results show that D. eres exhibited a high level of intraspecific variability within isolates, given that the strain CBS 160.32 might be a minor pathogen on blueberry plants, whereas CAA829 was revealed to be the most aggressive. Overall, this study also demonstrates that Diaporthe amygdali and D. eres may be two of the most aggressive species to blueberry plants. This study improves our understanding of the Diaporthe species and it's causing of dieback and twig blight on Portuguese blueberry orchards. Additionally, the identification of these pathogens represents crucial information for blueberry producers to apply appropriate phytosanitary measures, as well as offering new insights into the potential pathogenicity of D. eres on this host.