Diversity of wild Morchella (Ascomycota) revealed by integrated morphological and molecular analyses.

Morels (Morchella spp.) are highly valued edible and medicinal ascomycetes in temperate ecosystems, yet their taxonomy remains difficult because of marked morphological plasticity and the limited resolving power of single-locus markers. This study documents wild Morchella diversity from the Murree Hills, Pakistan, using an integrated morpho-cultural and molecular approach. Fifteen ascocarps collected from six localities during July-August 2024 were used for tissue isolation, yielding thirty-nine cultures, with multiple cultures recovered from individual ascocarps as independent isolation replicates. From these, nine representative isolates were selected for ITS-based sequencing and phylogenetic assessment. Culture recovery averaged 2.60 isolates per ascocarp, indicating successful establishment of cultures across sampling sites. Morphological and cultural observations supported the recognition of two principal forms: a pale morphotype assignable to the Esculenta clade (cf. Morchella deliciosa) and a darker morphotype affiliated with the Elata clade (within the Morchella elata complex). Bayesian ITS phylogeny and sequence-similarity comparisons supported this two-lineage pattern at the clade level. Because ITS alone is insufficient for confident delimitation of closely related Morchella species, these results should be regarded as preliminary lineage-level documentation rather than definitive species resolution. Nevertheless, this study provides baseline evidence for the occurrence of Esculenta- and Elata-affiliated morels in an understudied Himalayan region and establishes a foundation for future multilocus, population-level, and ecological investigations.