The diversity of Botryosphaeriaceae species associated with “Botryosphaeria dieback” of grapevine was investigated in 18 vineyards in Sardinia, Italy. Lasiodiplodia isolates obtained from different woody hosts including holm oak, sweet orange and broom bush in Italy, Algeria and Tunisia were also characterized. Morphological and cultural characteristics as well as ITS and EF1-α sequence data were used to identify the fungal isolates. Forty-eight botryosphaeriaceous isolates were obtained from 113 symptomatic grapevine samples, from which ten species were identified. Diplodia seriata was the dominant species (25 % of isolates), followed by Neofusicoccum parvum (21.7 %). Two species, Diplodia olivarum and D. africana are reported for the first time on grapevine. In addition, two new species namely Lasiodiplodia mediterranea sp. nov. from grapevine, holm oak and sweet orange and Lasiodiplodia exigua sp. nov. from broom bush are described. In artificial inoculation experiments conducted on excised green grapevine shoots and lignified canes as well as holm oak seedlings, L. mediterranea was shown to be an aggressive pathogen.
Studies on the taxonomy and phylogeny of Diplodia have been hampered by the lack of an ex-type culture linked to the holotype of D. mutila, which is the type of the genus. In this study a large collection of Diplodia strains, obtained from ash and other woody hosts showing V-shaped cankers and branch dieback, were identified based on morphological characters and DNA sequence data from ITS and EF1-α loci. Results of combined morphological and phylogenetic analyses showed that the Fraxinus isolates from Italy, the Netherlands, Portugal and Spain belong to three distinct species namely Diplodia fraxini, Diplodia mutila and Diplodia subglobosa sp. nov. An epitype was designated for Diplodia mutila, with associated ex-epitype cultures. The name D. fraxini is re-instated and a neotype designated. Two species, Diplodia seriata and Diplodia pseudoseriata were reported for the first time on Fraxinus spp.
Monitoring surveys of Phytophthora related diseases in four forest nurseries in Italy revealed the occurrence of fourteen Phytophthora species to be associated with collar and root rot on fourteen plants typical of Mediterranean and alpine regions. In addition, a multilocus phylogeny analysis based on nuclear ITS and ß-tubulin and mitochondrial cox1 sequences, as well as micromorphological features, supported the description of a new species belonging to the phylogenetic clade 7c, Phytophthora mediterranea sp. nov. Phytophthora mediterranea was shown to be associated with collar and root rot symptoms on myrtle seedlings. Phylogenetically, P. mediterranea is closely related to P. cinnamomi but the two species differ in 87 nucleotides in the three studied DNA regions. Morphologically P. mediterranea can be easily distinguished from P. cinnamomi on the basis of its smaller sporangia, colony growth pattern and higher optimum and maximum temperature values. Data from the pathogenicity test showed that P. mediterranea has the potential to threaten the native Mediterranean maquis vegetation. Finally, the discovery of P. cinnamomi in alpine nurseries, confirms the progressive expansion of this species towards cold environments, probably driven by climate change.