As a contribution to the lichen flora, two taxa of lichens are reported for the first time. The critical morpho-anatomical comparison and molecular phylogenetic construction based on Internal Transcribed Spacer (ITS-5.8-ITS2) barcode region revealed the identity of two lichenized Ascomycetes Leprocaulon nicholsiae and Candelaria asiatica from Margalla Hills of Pakistan. Their existence here indicates their first contemporary report from South Asia. We present the taxonomic characters, ecology, distribution along with an ITS-based phylogenetic trees of the newly collected specimens. Margalla Hills, Pakistan, DNA barcoding, lichenized ascomycetes, hills national-park, Life Sciences & Biomedicine - Other Topics