Multiple Botryosphaeriaceae and Phytophthora Species Involved in the Etiology of Holm Oak (Quercus ilex L.) Decline in Southern Italy

In recent years, severe decline and mortality events have been observed in holm oak (Quercus ilex L.) ecosystems in different Italian regions, including Puglia (southern Italy). Given the landscape and ecological relevance of holm oak forests in Apulia, a study was conducted to identify the causal agents related to this complex disease syndrome. The surveys, conducted in winter 2024 in three different woodlands, revealed the widespread occurrence of mature holm oak trees showing sudden death, crown thinning, shoot and branch dieback, sunken cankers, and root rot symptoms. Isolations performed from symptomatic samples collected from both stem and small roots yielded fungal and fungal-like colonies representing two distinct families: Botryosphaeriaceae and Peronosporaceae. Analysis of morphological and DNA sequence data allowed us to identify six distinct species, including Diplodia corticola and D. quercivora (Botryosphaeriaceae), Phytophthora cinnamomi, P. multivora, P. psychrophila, and P. asparagi (Peronosporaceae). For P. asparagi and P. psychrophila, isolated for the first time from declining holm oak trees in Italy, Koch’s postulates were satisfied by inoculating 1-year-old seedlings at the collar in controlled conditions. Thirty days after inoculation, all plants showed the same symptoms observed in the field. Overall, the data obtained highlights the co-occurrence of multiple Botryosphaeriaceae and Phytophthora species on declining holm oak trees and the discovery of a new haplotype of Diplodia quercivora.