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Oaks are dominant and key tree species in Mediterranean forest ecosystems. However, in recent decades, oak forests have been heavily impacted by oak decline, a worldwide phenomenon exacerbated by climate change. The charcoal disease agent Biscogniauxia mediterranea is involved in the decline of Mediterranean oak formations in a variety of contexts. Here, we investigated the impact and role of B. mediterranea in the decline of oaks in Castelfidardo Forest, a relict wood of the late Holocene and a Site of Community Importance. We established five plots within which we recorded tree positions, any symptoms and signs of decline, association of B. mediterranea to declining trees, and deadwood and associated mycota. Of 471 oaks inspected, 7.0% showed brownish exudates on the stems, 46.9% showed epicormic shoots along the main trunk, and 24.4% showed black carbonaceous stromata on diseased branches and trunks. The decline was most severe for Quercus cerris, which comprised plots #4 and #5, at 50.0% (81/162 trees) and 29.0% (33/114), respectively, then for Quercus robur for plot #3, at 40.0% (38/95), and finally for Quercus pubescens for plots #1 and #2, at 13.7% (7/51) and 12.3% (6/49), respectively. Bark tissue was collected from trees with charcoal cankers via microscopy examination and identified by mycological and molecular methods. This investigation revealed a close association between oaks with pronounced reduction of vitality and incidence of B. mediterranea. Deadwood was equally distributed among the five plots, and was heavily colonized by Basidiomycota. The high incidence of the charcoal canker pathogen B. mediterranea appeared to be related to environmental stresses. However, the absence of silvicultural management, high competition among physiologically mature trees, and the geographic isolation of this residual forest may have predisposed oaks to decline.

Oaks are a dominant and key tree species in Mediterranean forest ecosystems. However, in recent decades, oak forests have been heavily impacted by oak decline, a worldwide phenomenon exacerbated by climate change. The charcoal disease agent Biscogniauxia mediterranea is involved in the decline of Mediterranean oak formations in a variety of contexts. Here, we investigated the impact and role of B. mediterranea in decline of oaks in Castelfidardo Forest, a relict wood of the late Holocene and a Site of Community Importance. We established five plots within which we recorded the tree positions, any symptoms and signs of decline, association of B. mediterranea to declining trees, and the deadwood and associated mycota. Of 471 oaks inspected, 7.0% showed brownish exudates on the stems, 46.9% showed epicormic shoots along the main trunk, and 24.4% showed black carbonaceous stromata on diseased branches and trunks. The decline was most severe for Quercus cerris, which comprised plots #4 and #5, at 50.0% (81/162 trees) and 29.0% (33/114), respectively; then for Quercus robur for plot #3, at 40.0% (38/95); and finally for Quercus pubescens for plots #1 and #2, at 13.7% (7/51) and 12.3% (6/49), respectively. Bark tissue was collected from trees with charcoal cankers, with microscopy examination and identification by mycological and molecular methods. This investigation revealed close association between oaks with pronounced reduction of vitality and incidence of B. mediterranea. The deadwood was equally distributed among the five plots, and was heavily colonized by Basidiomycota. The high incidence of this oak-weakening pathogen B. mediterranea appears to be related to stress conditions (e.g., water scarcity, prolonged drought). However, absence of silvicultural management, high competition among physiologically mature trees, and the geographic isolation of this residual forest might have predisposed these oaks to decline.

Castanea sativa is a widespread and important multi-purpose tree in the Mediterranean area. Recently, intensive infestation of gall wasp decreased the production of chestnut, and makes the plants more susceptible to Cryphonectria parasitica, the causal agent of chestnut blight. Generally, biological control of chestnut blight is through natural spread or artificial release of hypovirulent strains of C. parasitica. However, this approach is not always successful. The aim here was to evaluate alternative approaches for control chestnut blight based on application of antagonistic microorganisms. In-vitro antagonistic activities of fungal species Trichoderma and Penicillium were tested after in vitro isolation from chestnut bark. In dual cultures, these antagonistic fungi consistently reduced growth of C. parasitica. Three commercial formulations based on Trichoderma spp., Glomus spp. and Bacillus subtilis were tested at three doses, both in vitro and directly applied to cuttings of chestnut stems as co-inoculations and as separated inoculations with C. parasitica mycelium plugs. With co-inoculation, B. subtilis showed the best performance against chestnut blight, with necrotic areas reduced by >70%. In separated inoculations, B. subtilis reduced the size of cankers on chestnut stems, by 29%–67%, as also for Trichoderma spp., by 36%–65%, and Glomus spp., by 31%–63%. Application of commercial formulations based on antagonistic microorganisms are effective tools in management of chestnut blight.

Grapevine trunk diseases (GTDs) are the most widespread fungal diseases, affecting grapevines in all the major growing regions of the world, and their complete eradication is still not possible. Aiming to search alternatives to avoid the spread and high incidence of these diseases, the present work intended to molecularly identify the grapevine endophytic community, the phytopathogenic fungi associated with GTDs in vineyards within the Alentejo region, and to test potential antagonist microorganisms as biological control candidates against GTDs-associated fungi. Grapevine endophytic community showed a wide variety of fungi in GTDs&rsquo, asymptomatic and symptomatic plants, nine of them previously described as GTDs-associated fungi. GTDs prevalent fungi identified in symptomatic plants were Diaporthe sp., Neofusicoccum sp., and H. viticola. Almost all these fungi were also detected in asymptomatic plants, which shows the importance of investigating the interactions of fungal communities and confirms the need for early diagnosis of these diseases. Direct inhibition antagonism tests were performed among identified endophytes and GTDs phytopathogenic fungi, and all the endophyte fungi showed potential as biocontrol agents. Our findings suggest that endophytes are promising candidates for their use in biological control due to their antagonistic activity against the mycelia growth of some GTDs-associated fungi.