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A new lichen species Lecaimmeria pakistanica K.Habib, R.Zulfiqar & Khalid sp. nov. is described and illustrated from rocks in the temperate forests of the Himalaya of Azad Jammu and Kashmir, Pakistan. This species is characterized by its yellow-brown to brown thallus having areoles 0.4 to 1.5 mm across, branched and anastomosing paraphyses, a tall hymenium, large ascospores 20–32 × 10–16 μm, and no substance detected by thin layer chromatography. All other species of the genus have ascospore dimensions in the range of 14–22 × 5–14 μm. A phylogenetic analysis is provided based on ITS nrDNA sequences, and supports the separation of the novel species. Photographs and a comparative analysis with related species of Lecaimmeria are provided to confirm the status of the species.

An updated key to the species of Oxneriaria 1 Non - zonate, distinctly areolate, grey to whitish grey upper surface, and prothallus absent, apothecia 0.5– 2 mm........................................................................................................................................................................................ Oxneriaria pakistanica sp. nov. 2 Usually lacks distinct areoles, up to 2 mm apothecia, prothallus present......................................................... Oxneriaria rivulicola 3 Often elongated, giving thallus subplacodoid appearance, 0.2–0.5 mm apothecia, black prothallus, 70–80 µm tall hymenium.............................................................................................................................................................................. Oxneriaria verruculosa 4 Elongated marginal areoles, upto 0.6 mm apothecia, prothallus present..................................................... Oxneriaria dendroplaca 5 Whitish mealy thallus, larger ascospores 20–24×15–20 µm......................................................................... Oxneriaria nikrapensis, Published as part of Iqbal, Muhammad Shahid, Usman, Muhammad, Habib, Kamran & Khalid, Abdul Nasir, 2023, Oxneriaria pakistanica sp. nov. (Megasporaceae, Pertusariales, Ascomycota) from Darel Valley, Gilgit Baltistan, Pakistan, pp. 125-131 in Phytotaxa 579 (2) on page 130, DOI: 10.11646/phytotaxa.579.2.6, http://zenodo.org/record/7543142

A new species in Megasporaceae, Oxneriaria pakistanica is described and illustrated from Pakistan. A comparative morpho–anatomical study and ITS–based molecular analyses confirmed its position within the recently resurrected genus, Oxneriaria. The taxon has non-zonate thallus, epruinose, distinctly areolate, 0.5–1.8 mm wide areole, grey to whitish grey at upper surface, prothallus absent, 0.1–2 mm in diameter apothecia, taller hymenium 100–155 µm, deeper hypothecium 90–170 µm, broadly ellipsoid to sub-spherical ascospores 10–18 × 7–10 µm size and chemistry unknown substance detected. Its forms a separate branch in the phylogenetic tree and also distinct from the other known taxa of the genus. Oxneriaria pakistanica sp. nov appeared to be a sister specie to Oxneriaria rivulicola. A detailed comparison and a key to all taxa of Oxneriaria is also given.

Oxneriaria pakistanica M.S Iqbal, Usman, K. Habib, Khalid sp. nov. MYCOBANK MB 845095 Etymology: The epithet ‘ pakistanica ’ refers to the type locality of the country Pakistan. Diagnosis: Oxneriaria pakistanica is differs from its closely related species Oxneriaria rivulicola by having nonzonate, distinctly areolate, grey to whitish grey upper surface, and prothallus absent, immersed apothecia (1–1.5mm), broadly ellipsoid to sub-spherical ascospores (10–18 × 7–10 µm). Holotype:— PAKISTAN: Gilgit Baltistan, Darel Valley 35° 37’N, 73° 27’E, elev. 1,900 m, on rocks, 21 October 2020, Muhammad Shahid Iqbal DR-42 (LAH37495), (ITS GenBank accession number OP114649). Description: Thallus: crustose, non zonate, inially placodoid (lobate), becoming non placodoid with age, 5 cm across. Areoles: distinctly areolate, uncracked, 0.5–1.8 mm wide, minute at ages, broadly attached, discrete to rarely contiguous, thinly wrinkled, irregular or angular to irregular elongated, flat to strongly convex. Upper surface: epruinose, dull, whitish grey to grayish, unchanged when wet. Prothallus: not found. Upper Cortex: paraplectenchymatous, two-layered, above brown, below hyaline, 10–25 µm thick, cells 5–10 µm in diam. Algal layer: discontinuous, 30–50 µm thick, photobiont chlorococcoid, cells ± spherical, 10–15 µm in diam. Medulla:, hyphae prosoplectenchymatous, 3–5 µm wide. Apothecia: frequent, aspiciloid, immersed, covering whole areole, 1–3 per areole, mostly one per areole, become confluent many, rounded to angular, 0.5–2 mm in diam. Disc: 0.5–2 mm in diam, black, dull, plane to weakly concave, irregular to elongated, often with depressions, epruinose. Margins: concolorous to thallus, elevated, thick, continuous, thinly wrinkled. Thalline exciple: 90–120 µm wide. Proper exciple: indistinct. Epihymenium: brown to dark brownish, 20–30 µm thick. Hymenium: hyaline, 100–155 µm tall. Hypothecium: hyaline, 90–170 µm deep. Paraphyses: moniliform, septate, 2–4 celled capitate, cell 3.5–5.5 µm wide elevated. Asci: clavate, 8-spored, 60–80 × 30–40 µm. Ascospores: simple, hyaline, broadly ellipsoid to sub-spherical, 10–18 × 7–10 µm. Chemistry: Cortex K+ (yellowish green), C-, KC + (light green) Medulla: all negative TLC: Unknown substance detected Habit and Habitat: The recognized collections of new species are from a moist temperate climate, in an open situation exposed to sun and rain, found at the hilly topography of Darel valley with an altitude of 3.843m. In area, summers are warm and clear and the winters are freezing, snowy, and partly cloudy. The specimens were on calcareous sedimentary rocks. Common floral species are trees and shrubs. Pinus gerardiana, Cedrus deodara, Pinus wallichiana, Fraxinus xanthoxyloides. The average annual precipitation in the valley is 100–300 mm, mostly occurring during winter and early spring in the form of snow. Mean temperatures range from -10 °C in winter to +35 °C in summer. Additional Specimen:— PAKISTAN: Gilgit Baltistan, Darel Valley 36° 38’N, 74° 28’E, elev. 2,000 m, on rocks, 10 August 2022, Muhammad Shahid Iqbal DR-106 (LAH 37501), (ITS GenBank accession number OP627196). Comments Morphologically, Oxneriaria pakistanica can be distinguished from the superficially similar taxon Oxneriaria verruculosa in having 0.5–1.8 mm wide areoles (vs 0.5–1mm wide), distinctly areolate (vs often elongate, giving thallus subplacodoid appearance), epruinose (vs. grey pruinose), prothallus absent (vs. sometimes delimited by black prothallus), 0.5–2 mm in diam apothecia (vs. 0.2–0.5 mm), taller hymenium 100–155 µm (vs. 70–80), deeper hypothecium 90–170 µm (vs 20–30 µm), larger ascospores 10–18×7–10 µm (vs 12–17×7.5–9 µm), chemistry unknown substance detected (vs stictic acid, rarely norstictic acid) (Nimis 2016). Phylogenetically, Oxneriaria rivulicola is close to the Pakistani taxon but can be easil differentiated as the former has zonate thallus, usually lacks distinct aeroles but with radiating cracks, often with a brownish ting and prothallus usually present, apothecia up to 2 mm, chemistry: no substance detected. Nordin et al., (2011). The Pakistani taxon is non - zonate, distinctly areolate, grey to whitish grey upper surface, and prothallus absent, apothecia 0.5–2 mm, chemistry: unknown substance detected. Another morphological similar taxon to Oxneriaria pakistanica is Oxneriaria dendroplaca, which has also distinctly - areolate thallus but has black Prothallus, elongated marginal areoles, smaller apothecia ( upto 0.6 mm ), and brown to dark greenish grey thallus, size of ascospores 13.6–[15.7]–19.2×7.9–[8.9]–11.9, whereas the Oxneriaria pakistanica has none elongated marginal areoles, prothallus absent, larger apothecia 1–2 mm and whitish gray to gray thallus, larger ascospores 10–18×7–10 µm.Nordin et al., (2011). The ITS nrDNA also support the separation of Oxneriaria pakistanica in phylogenetically. The sequence of Oxneriaria nikrapensis is not available in Genbank. Oxneriaria nikrapensis whitish mealy thallus, larger ascospores 20–24 × 15–20 µm and stictic acid detected by TLC. Chesnokov et al., (2018)., Published as part of Iqbal, Muhammad Shahid, Usman, Muhammad, Habib, Kamran & Khalid, Abdul Nasir, 2023, Oxneriaria pakistanica sp. nov. (Megasporaceae, Pertusariales, Ascomycota) from Darel Valley, Gilgit Baltistan, Pakistan, pp. 125-131 in Phytotaxa 579 (2) on pages 128-130, DOI: 10.11646/phytotaxa.579.2.6, http://zenodo.org/record/7543142, {"references":["Nimis, P. L. (2016) ITALIC - the information system on Italian lichens. Versions 5.0 University of Trieste. Dept. of Biology. [http: // dryades. units. it / italic]"]}

Lecaimmeria pakistanica K.Habib, R.Zulfiqar & Khalid sp. nov. MB844738 Fig. 2 Diagnosis Distinguished from all the known species of the genus by having large ascospores (20–32 × 10–16 μm), and relatively taller hymenium. All the other species of the genus have ascospore dimensions in the range of 14–22 × 5–14 μm. Also separated from other species of the genus by ITS nrDNA sequence data. Etymology The specific epithet ʻ pakistanica ʼ refers to country in which the new species was discovered. Material examined Holotype PAKISTAN • Azad Jammu and Kashmir, Muzaffarabad, Peer Chinasi; 34°23′ N, 73°32′ E; alt. 2924 m; on rocks; 9Aug. 2018; T. Saifullah and K. Habib leg.; PC-21; LAH [LAH-36674]; GenBank no.: MW508503. Paratype PAKISTAN • Azad Jammu and Kashmir, Muzaffarabad, Peer Chinasi; 34°23′ N, 73°32′ E; alt. 2700 m, on rocks; 22 Jul. 2019; T. Saifullah and K. Habib leg.; PC-22; LAH [LAH-36675]; GenBank no.: MW508504. Description Thallus crustose, areolate, up to 6 cm wide, in section 200–280 μm thick, upper surface yellow-brown to brown, no change when wet. Areoles separate, flat to weakly convex, irregular to angular, slightly pruinose near margin, glossy, adnate, without fissures, marginal areoles slightly larger, up to 1.5 mm across, up to 0.6 mm thick, rarely with whitish margins. Prothallus visible between areoles, blackish. Cortex two layered, ca 40–60 μm thick, paraplectenchymatous, cells 8–12 μm in diam., upper layer paler brown, 10–16 μm thick, lower layer hyaline, 30–40 μm thick, epinecral layer distinct, up to 20 μm high. Algal layer 80–120 μm thick, chlorococcoid, cells globose to subglobose, 10–20 μm in diam. Medulla: hyphae white, 15–30 μm thick, corresponding with areole, IKI+ blue. Apothecia crypto-lecanorine, frequent at center of thallus, 1–4 per areole, immersed, sometimes not completely surrounded by the areole. Disc contiguous to separate, flat to concave, reddish brown, rounded at first becoming irregular, sometimes surrounded by a white rim, up to 0.8 mm diam., thinly to rarely pruinose, margin pruinose. Proper exciple thin, poorly differentiated, reduced, hyaline, 10–25 μm thick. Hymenium hyaline, 130–160 μm tall including the epihymenium, which is pale brown to brown, 10– 15 μm thick, epinecral layer 3–7 μm thick; paraphyses apically branched, anastomosing, 1–2 μm wide, apically slight swollen, apices 2.5–3.5 μm wide. Hypothecium 60–100 μm tall, light grayish brown, containing algal cell in the lower part. Asci Porpidia - type, clavate, 80–130 × 25–45 μm, amyloid wall 4–6 μm thick, 8-spored; ascospores hyaline, ellipsoid to broadly ellipsoid, mature with germ tube, 20–32 × 10–16 μm. Spot tests cortex and medulla K-, C-, KC-; medulla IKI+ blue; TLC none detected. Ecology Growing on sun-exposed rocks in a dense forest at an altitude of 2900 m. Topography is mountainous in Himalayan region. Dominant tree species are Pinus roxburghii Sarg., Pinus wallichiana A.B.Jacks., Cedrus deodara (Roxb.) G.Don, Picea smithiana (Wall.) Boiss, Abies pindrow Royle. Maximum and minimum temperature of 32°C and -8°C, respectively. Annual rainfall varying between 1000–1500 mm., Published as part of Habib, Kamran, Zulfiqar, Rizwana & Khalid, Abdul Nasir, 2022, Lecaimmeria pakistanica, a new lichen from Azad Jammu and Kashmir, Pakistan, pp. 94-101 in European Journal of Taxonomy 834 on pages 96-99, DOI: 10.5852/ejt.2022.834.1901, http://zenodo.org/record/7010513

Aspiciliella pakistanica K. Habib, Q. Firdous, Sohrabi & Khalid MycoBank No: 838033 Diagnosis: The taxon is characterized by a whitish���grey thallus, 0.8���1.5 mm thick, with areoles 1���4.2 mm wide without pale lines on the surface and a discontinuous algal layer arranged in groups of vertical rows. Etymology: The specific epithet pakistanica (Latin) refers to the country of origin of this species Typus: PAKISTAN. Azad Jammu & Kashmir: Muzaffarabad, Pir Chinasi, 34�� 23��� 20.76��� N, 73�� 32��� 59.64��� E, 2,900 m a.s.l.; on rocks; August 3, 2018, T. S. Ullah & K. Habib PC ���15 (LAH 36686 ��� Holotype) and Isotype (ICH ��� Isotype). Thallus: crustose, areolate to rimose areolate, 5 to 8 cm wide, epruinose. Areoles: 1���4.2 mm wide and 0.8���1.5 mm thick, thicker centrally but thinner toward the periphery, broadly attached to stipitate, separated by distinct cracks, discrete to rarely contiguous, smooth to thinly wrinkled, angular or irregular, replicating by division, flat to weakly convex. Upper surface whitish���grey in the field, becoming pale yellow-brown to brown in the herbarium. Lower surface dark brown to black. Prothallus: not apparent. Upper Cortex: 10���20 ��m thick, smooth, upper layer dark gray���brown, hyaline adjacent to the algal layer, paraplectenchymtous, textura globularis, cells 5���10 ��m in diam. Algal layer: discontinuous, clustered in groups, arranged in vertical rows; rows 30���50 ��m wide and 65���120 ��m high, hyaline between the gaps. Photobiont chlorococcoid, cells �� spherical, 10���15 ��m in diam. Medulla: 300���570 ��m thick, corresponding with size of the areole, hyphae prosoplectenchymatous, 3���5 ��m wide. Pycnidia: immersed, rare, colorless, indistinct, 300���380 ��m high �� 140���230 ��m wide. Conidiospores: 15���25 �� 1���2.5 ��m. Conidia: acrogenous, filiform, straight, hyaline, 8���12 �� 0.5���1 ��m. Spot tests: Cortex; K+ (yellowish���red), C���, KC���. Medulla; K���, C���, KC���. Ecology: Saxicolous in a temperate climate at an altitude of 2900 m a.s.l., rain and sun exposed, with surrounding trees Abies pindrow (Royle ex D.Don)Royle, Cedrus deodara (Roxb.) G.Don, Pinus roxburghii Sarg., Pinus wallichiana Jacks and Picea smithiana, (Wall.) Boiss dominant and hilly topography, having a maximum and minimum temperature of 34 oC and ���3 oC, respectively and annual rainfall varying from 1400���1800 mm. The second collection was found at an elevation of 3000 m a.s.l in the partially forested Kaghan valley, with mean maximum and minimum temperature of 32��C and ���2��C, respectively, and temperate climate. It is a biologically, geo���physically, and hydrologically dynamic area. Additional Specimen Examined (Paratype): PAKISTAN. Khyber Pakhtunkhwa: Kaghan; 34.5417�� N, 73.3500�� E; 3000 m a.s.l; on rocks; July 19, 2019, A. N. Khalid & K. Habib; K ��� 23 (LAH 36686)., Published as part of Habib, Kamran, Firdous, Qudsia, Sohrabi, Mohammad & Khalid, Abdul Nasir, 2021, Aspiciliella pakistanica a new lichen species (Megasporaceae, Pertusariales, Ascomycota) from Pakistan, pp. 175-182 in Phytotaxa 511 (2) on pages 179-180, DOI: 10.11646/phytotaxa.511.2.5, http://zenodo.org/record/5426644