Abstract
English walnut (Juglans regia) is a species that is highly valued for the quality of its wood and the nutritional and nutraceutical properties of its fruit. A severe dieback of J. regia trees was observed recently in orchards located in three geographically distinct areas of Tuscany, central Italy. Symptoms included root and collar rot, necrosis of the under-bark tissue, bleeding cankers, stunted growth, and crown dieback. Four Phytophthora species were obtained from 239 isolates found on symptomatic J. regia individuals. They were identified, on the basis of macro-morphological (colony shape and texture), micro-morphometric (shape and size of oogonia, antheridia, oospores, sporangia, and chlamydospores) and molecular (ITS sequencing) characters, as P. gonapodyides, P. cactorum, P. citricola, and P. plurivora. Among these species, P. plurivora was the species isolated with overwhelming frequency from symptomatic tissue and rhizosphere soil, suggesting it to be the putative etiological agent. Pathogenicity assays were conducted on 20 cm long detached J. regia branches for a definitive establishment of disease causation. Severe symptoms (extended necroses) were exhibited by branches infected with P. plurivora, proving its pathogenicity and high virulence on this host. The other Phytophtora species produced negligible necroses around the inoculation site. P. plurivora was recovered from all the investigated orchards, providing evidence that it is quite widespread. This study highlights the growing threat posed by the polyphagous P. plurivora to walnut cultivation and the sustainable business it fuels in Europe, underscoring the need for integrated management strategies to mitigate its economic and ecological impacts.