Evernia prunastri Ach.

Evernia prunastri (L.) Ach., Lich. Univ.: 442, tab. 10, fig. 1 (1810) Fig. 3 Description. Thallus foliose but appearing fruticose, dorsiventral, tufted, caespitose to occasionally pendent, cortex cracked at some places, palmately to dichotomously branched, up to 5.5 cm across. Branches: numerous, dorsiventral, elongated, 0.3–4 mm wide, plane, strap-shaped, pitted, twisted, tangled, blunt or pointed. Axils: right angles to V-shaped. Color: greyish-green to greenish-white when dry, pale olive green to yellow-green when wet. Soredia: marginal to rarely laminal, white, often on ridges, granular. Upper surface: matt, dull, pitted, strongly rugose, rough, ridges discontinuous. Lower surface: white to pale yellow, channelled, less pitted and wrinkled than upper surface. Cortex: two-layered, upper layer brown, 9–14 µm high, lower layer hyaline, 7–12 µm high, paraplectenchymatous, cell 7–12 µm in diameter. Medulla: prosoplectenchymatous, hyphae loosely arranged, white, 2–4.5 µm wide. Algal layer: discontinuous, uneven, forms small groups, algal cells present below cortex, 5–10 µm in diameter. Apothecia: not observed. Chemical study. K + ve (yellow), C - ve, KC + ve (yellow). Usnic acid, atranorin and evernic acid detected through TLC. Phylogenetic analysis. For phylogenetic analysis, sequences of the genus Evernia were retrieved from GenBank based on similarity and those used in a study by Piercey-Normore (2006). A total of 22 ITS nrDNA sequences were analyzed during this study. The final aligned file was comprised of 532 nucleotides, among which 359 were conserved, 158 were variable, 150 were parsimony-informative, and 8 were singleton variants. Umbilicaria kappenii Sancho, B. Schroet. & Vallad. (AY 603130 and AY 603131) was chosen as an outgroup. In our analysis, Evernia prunastri forms a distinct clade and the ITS sequences of E. prunastri (KUM- 04 and KUM- 09) from Pakistan are identical to those reported from Norway (MK 811838, MK 811993, MK 812247, and MK 812058), confirming their identification as the same species (Fig. 4). Additionally, E. divaricata was distinctly separated from E. mesomorpha and E. esorediosa in the phylogenetic tree, yet all three species formed a clade with a high bootstrap support (92 % ML). This study represents the first molecular confirmation of E. prunastri from Pakistan. Notes. Evernia prunastri, commonly known as oakmoss, is a fruticose lichen found predominantly on exposed tree trunks, characterized by flat, isotomic-dichotomous branches with frequent anisotomous bifurcations and occasional adventive branches or “ lobuli ” on the branch surface (Beltman 1978; Stone and McCune 1990). The morpho-anatomical comparison of Pakistani collections confirms their identity as E. prunastri, as the characteristics of Pakistani samples match with the already published description of E. prunastri from Britain and Ireland (Cannon et al. 2023). Habitat and ecology. The samples were found at an elevation above 2000 m, growing on bark in the cold temperate mountainous regions. The coldest month is December, while June is the hottest month of the year, with average monthly temperatures of 3 ° C and 25 ° C, respectively, and rainfall varying between 600 and 1200 mm. The dominant vegetation includes Indian horse-chestnut (Aesculus indica (Wall. ex Cambess.) Hook.), deodar (Cedrus deodara (Roxb. ex D. Don) G. Don) and West-Himalayan fir (Abies pindrow (Royle ex D. Don) Royle). Distribution. This species is widely distributed in Northern hemisphere including Asia, Europe and North America (Studzińska-Sroka et al. 2023; Staples et al. 2020). Materials examined. Pakistan ∙ Khyber Pakhtunkhwa Province, District Upper Dir, Kumrat Valley, 35°23.2'N, 74°35.0'E; alt. 2,500 m a. s. l.; 23 Sep. 2023; M. Usman & A. N. Khalid leg.; on bark; KUM- 4, ITS GenBank accession number PQ 113696 ∙ same collection data as for preceding, 35°33.9'N, 72°11.4'E; alt. 2,400 m a. s. l.; 23 Sep. 2023; M. Usman & A. N. Khalid leg.; on bark; KUM- 9, ITS GenBank accession number PQ 113697.