Associations Between the Leaf Microbiome and the Health of Irish Ash Trees Affected by Hymenoscyphus fraxineus

Ash dieback, caused by the fungus Hymenoscyphus fraxineus, continues to threaten European ash (Fraxinus excelsior), yet the contribution of the leaf microbiome to disease severity remains poorly understood. We surveyed 133 ash trees across nine sites in Northern Ireland, using canopy cover as a proxy for health, and characterised leaf-associated microbial communities using Pacific Biosciences (PacBio) long-read amplicon sequencing (full-length 16S and ITS) and QIIME2-based workflows. Many trees exhibited partial tolerance to ash dieback, with most maintaining stable canopy cover year-on-year, while fewer trees show a decline and a smaller portion showing improvement. Microbial communities were largely stable irrespective of ash health with little difference in alpha diversity (Shannon) or beta diversity (Bray–Curtis PERMANOVA) for either bacteria or fungi. Differential abundance and correlation analyses showed that H. fraxineus was, as expected, negatively associated with canopy cover. Only one fungal species, Papiliotrema flavescens, demonstrated a strong positive association with healthier trees, consistent with previous findings. These results indicate that Northern Ireland hosts a reservoir of ash trees displaying tolerance to ash dieback. While the leaf microbiome does not appear to drive this tolerance at the community level, one fungus, P. flavescens, was correlated with healthier ash.