Genetic diversity of true morels (Morchella) in Pakistan inferred from multilocus DNA sequence data augmented by a retrospective analysis of Pakistani and Indian morel sequences in GenBank

The current study was conducted to obtain the first detailed assessment of true morel (Morchella) species diversity in Pakistan. A collection of 179 morels was obtained by conducting field surveys in 13 districts during spring and autumn of 2014 through 2020. Species were identified phylogenetically by analyzing portions of two (RPB1, RPB2) or four (RPB1, RPB2, TEF1, ITS rDNA) marker loci together with authentic reference sequences of known species and by conducting BLASTn queries of National Center for Biotechnology Information (NCBI) GenBank. A retrospective analysis of morel sequences from Pakistani (N = 52) and Indian (N = 69) collections accessioned in GenBank was also conducted to expand and compare our knowledge of Morchella species diversity in both countries. Analyses of our 179-morel collection and the 52 Pakistani sequences deposited in GenBank indicated that at least 20 species are present in Pakistan, including five putatively novel taxa within the Elata subclade here informally distinguished as Morchella spp. (Mel-43 to Mel-47). Phylospecies Mel-45 and Mel-46 were described formally as M. pycnogranulata and M. waziristanica, respectively. By comparison, analyses of the 69 Indian sequences deposited in GenBank indicated that at least 12 species are extant in India. However, only 4/9 Pakistani and 3/12 Indian species were correctly identified in GenBank. Two collections of the steppe morel, M. steppicola (Mes-1), from Afghanistan were also included in the present study, which extends the southeasternmost range of this morphologically distinct species.