Morphoanatomical and molecular characterization of two new Russula species in Section Ingratae from Pakistan Author Razzaq, Annum 0000-0002-3593-5821 Fungal Biology and Systematics Research Laboratory, Institute of Botany, University of the Punjab, Lahore, Pakistan. & annum. phd. botany @ pu. edu. pk; https: // orcid. org / 0000 - 0002 - 3593 - 5821 annum.phd.botany@pu.edu.pk Author Naseer, Arooj 0000-0002-4458-9043 Fungal Biology and Systematics Research Laboratory, Institute of Botany, University of the Punjab, Lahore, Pakistan. & arooj. hons @ pu. edu. pk; https: // orcid. org / 0000 - 0002 - 4458 - 9043 arooj.hons@pu.edu.pk Author Shahid, Soumia 0000-0001-7327-6495 Fungal Biology and Systematics Research Laboratory, Institute of Botany, University of the Punjab, Lahore, Pakistan. & somiashahid 03 @ gmail. com; https: // orcid. org / 0000 - 0001 - 7327 - 6495 somiashahid03@gmail.com Author Khalid, Abdul Nasir 0000-0002-5635-8031 Fungal Biology and Systematics Research Laboratory, Institute of Botany, University of the Punjab, Lahore, Pakistan. & drankhalid @ gmail. com; https: // orcid. org / 0000 - 0002 - 5635 - 8031 drankhalid@gmail.com text Phytotaxa 2023 2023-04-13 592 1 21 38 http://dx.doi.org/10.1094/PDIS-04-22-0755-PDN journal article 10.11646/phytotaxa.592.1.2 1179-3163 7835643 Russula bhurbanensis Razzaq, Naseer & Khalid sp. nov. Figures 3 & 4 MycoBank:—MB841965 Diagnosis:— Russula bhurbanensis is a medium sized species characterized by sticky, orange to dark brown, largesized pileus ( 7.5‒11.2 cm ); adnexed and crowded lamellae; amyloid and fairly large spore (7.1‒9.9×6.1‒9.2 µm) with isolated and occasionally fused warts with rare line connections; abundant lanceolate cheilocystidia with contents. Holotype :— Pakistan , Punjab , Rawalpindi , Bhurban , 33.9554° N , 73.4519° E , elev. 1828 m , on soil under Pinus wallichiana , solitary, 07 September 2020 , Annum Razzaq BR39 ( LAH36938 , GenBank accession number for ITS : MW886216 , GenBank accession number for LSU : MW886218 ). Etymology:—‘bhurbanensis’ is referring to the type locality Bhurban, Punjab , Pakistan . Description: — Basidiomata medium to large sized. Pileus 7.5‒11.2 cm diameter, applanate to plano-concave when immature becoming depressed to infundibuliform at maturity, margin tuberculate-striate, cuticle sticky to viscid, peeling to half of the radius; prominent radial striations at periphery, with smooth central disc, in young specimen orange (2.3Y 4.9/12.6) margins and dark brown (7.9YR 5.6/5.6) central disc, in mature specimen centrally orange (2.3Y 4.9/12.6) becoming reddish yellow (6.7YR 6.2/11.3) with yellow (2.6Y 8.6/4) towards margins, in old specimen dark brown (0.4YR 3.5/5.3) centrally with lighter shade of brown (7.9YR 5.6/5.6) towards margins. Lamellae well developed, creamy yellow (3.9Y 9.7/0.5) with brown spots (6YR 7.2/3.6), crowded, adnexed, rarely alternating with short lamellulae in single tier, subventricose to ventricose, edge even, entire and concolorous. Stipe 6.9–12× 1.8–2.2 cm , equal to subclavate, central, longitudinally striated, yellowish-brown (9.7YR/3.3) to dark brown (5YR 3.6/5.9) patches towards the lower half of the stipe, beige (6.6B 9.9/0.6) at base. Context thin in pileus, white, hollow in stipe. Taste not recorded. Odor strongly unpleasant. Spore print not recorded. Basidiospores [80/3/3] (7.1‒) 7.5‒8.7‒9.7 (‒9.9) × (6.1‒) 6.4‒7.6‒9.0 (‒9.2) µm, Q = 1.07‒1.13‒1.18, subglobose to broadly ellipsoid, ornamented, distant to moderately distant [(3–)4–5(–6) in a 3 µm diam. circle], mostly isolated, occasionally fused in pairs or triplets, rarely connected by fine line connections prominent hilar appendix, 0.85–2.12 µm, guttulated, amyloid. Basidia 43.4‒61.6×11.4‒15.8 µm, 4-spored, sterigmata avl =2.2‒5.5 µm, broadly clavate, content present. Basidiola 26.3 – 40.3×7.9 – 11.9 µm, cylindrical to clavate. Pleurocystidia 74.9 ‒ 119×7.7 ‒ 14.2 µm, mainly clavate, fusiform or lanceolate similar to cystidia on lamellae edge but relatively larger. Lamellar edges fertile. Cheilocystidia 72.4 ‒ 89.4×6.7 ‒ 12.2 µm, abundant, lanceolate or fusiform, apically acute to obtuse, mucronate 2‒8 µm long appendage, thin to moderately thick walled, contents crystalline like masses. Marginal cells 10.3‒20.5×4.8‒6.3 µm, small, subcylindrical to clavate. Hymenophoral trama mainly composed of numerous sphaerocytes measuring 19.3 – 29.7×15.6 – 23.4 µm. Pileipellis a mixture of cylindrical hyphae 2.26 – 8.14 μm and sphaerocytes 23.8 – 46.1×18.9 – 24.7 µm,, septate, branched, parallel in arrangement, thin walled, highly flexuous. Hyphal terminations near the pileus margin intricate, less flexuous, thin walled, terminal cells 22.7‒54.5×3.4 ‒ 9.4 µm, clavate with content. Hyphal terminations near the pileus center more flexuous, thin walled, terminal cells 18.7 ‒ 59.9×3.1‒8.1 µm, clavate, thin walled, heteromorphous content. Pileocystidia near pileus margins infrequent, 54.7‒94.4×3.2‒5.7 µm, 1‒3 celled, cylindrical, mostly clavate, apically obtuse to slightly constricted, dense content. Pileocystidia near pileus center 60.2‒91.4×2.5‒5.1 µm, more frequent, thin walled, heteromorphous content, apically obtuse to constricted, always 1- celled. Clamp connections absent in all tissues. FIGURE 3. Macromorphological characteristics of Russula bhurbanensis A – B = BR39 ( Holotype LAH36938 ) , C – D = GB24 (LAH36937). FIGURE 4. Microscopic characteristics of Russula bhurbanensis BR 39 ( Holotype LAH36938 ): A. Basidia; B. Basidioles; C. Pleurocystidia; D. Cheilocystidia; E. Pileocystidia (on left side) and hyphal terminations (on right side) at pileus center; F. Pileocystidia (on left side) and hyphal terminations (on right side) at pileus margin. Additional material examined: — Pakistan , Punjab , Rawalpindi , Bhurban , 33.9554° N , 73.4519° E , elev. 1828 m , on soil under coniferous tress, 04 September, 2020, Annum Razzaq BR97 ( LAH36939 ! , GenBank accession number for ITS : MW886217 , GenBank accession number for LSU : MZ343175 ) . 07 September , 2020, Annum Razzaq GB24 ( LAH36937 ! GenBank accession number for ITS : MZ686262 ) .