Taxonomic observations & Morphological characterization of Russula species
Russula pauriensis A. Ghosh, K. Das & Buyck, Cryptogamie, Mycologie 38 (3): 392 var. caulocystidii var. nov.:
Mycobank Number 822859 Fig. 1–6
Genbank Number MW 882089
Entomology The variety is named after the presence of bunches of regular tufts of caulocystidia.
Basidiocarps 8.0-9.5 cm in height. Pileus 6.0-6.2 cm in diameter, reflexed with depressed centre, umbo absent, surface grayish magenta (13A4) to dark purple (14F6) towards apex with greenish and yellow tings towards margins; dry; scaly, appressed fibrillose scales; glabrous; feebly striate; margins irregular, splitting at maturity; cuticle half peeling; flesh up to 0.6 cm broad, creamish white, unchanging. Pileal veil absent. Odour mild. Lamellae up to 0.8 cm broad, adnexed, equal, close, creamish white with yellowish white (1A2) tinge, forked near the attachment to stipe, unchanging; gill edges smooth. Stipe excentric, 7.0-7.4 cm long, 2.1 cm towards apex, 1.8 cm at centre and 1.8 cm at base, slightly tapering downward towards the base, creamish white with reddish lilac (14B5) tinges; solid; surface fibrillose; unchanging; exannulate.
Basidiospores (6.4) 7.47–8.3 × 6.6–7.47µm; Q = 0.96 (1.13)-1.1 µm; globose to sub-globose; warty, warts 0.8–1.6 µm high, isolated, catenulate, incomplete reticulum; amyloid; apiculate, apiculus 2.4–3.6 µm long. Basidia 38.4–65.6 × 11.2–12.8 µm, clavate, granular, tetra-sterigmate, sterigmata 4.0–8.0 µm, granular, pointed. Pleurocystidia 65.6–120.0 × 9.6–14.5 µm, fusiform with long tubular neck, blunt and acute apices, double walled, thick walled, granular, tip is filled with thick granulation, deeply embedded, abundant. Cheilocystidia 28.8–57.6 × 6.4–12.8 µm, fusoid ventricose with blunt and acute apices, thick walled, granular. Hymenophoral trama regular. Gill edges sterile. Pileus cuticle hyphal, ixocutis, made up of 4.8–8.8 µm broad, loosely arranged, septate, horizontally tangled hyphae giving rise to regular turf of broad, granular, septate hyphae, terminal hyphal cells subulate, ellipsoid to subfusoid; context hyphal made up of 3.2–8.0 µm broad, septate, granular hyphae intermixed with 4.8–52.8 µm, irregular, hyaline cells. Stipe cuticle hyphal, non-gelatinized, 3.2–7.2 µm broad, granular hyphae intermixed with 4.0–12.8 µm, granular, irregular cells, giving rise to regular tufts of bunches of caulocystidia; context cellular, made up of 14.4–40.0 µm broad, hyaline, irregular cells. Caulocystidia 56.0–72.8 × 5.6–8.0 µm, fusoid ventricose with blunt and acute apices, thick walled, densely granular. Clamp connections absent.
Chemical Reactions KOH: Dry pileus surface turns reddish orange in KOH.
Collection examined Narkanda, Kotgarh road (2084m) 31̊ 17’0ʺN − 77̊ 27’15ʺE, growing scattered on humicolous soil in mixed forest during late monsoon season, 14th October 2018, Amandeep Kaur, PUN 11187.
Result Morpho-antomically the present collection matches well with the description given for Russula pauriensis [14]. This collection differs as there is present a regular tuft of bunches of caulocystidia on the stipe cuticle which are all together lacking in R. pauriensis or its allied species like R. mukteshwarica [15]. Thus, new variety is being proposed as Russula pauriensis var. caulocystidii. R. pauriensis is recorded from study area for the first time and a new var. is proposed. Morpho-anatomical characters matches well with molecular characterization of Russula pauriensis. Molecular analysis gave 98% query cover and 98.94% percent identification with Russula pauriensis.
Phylogen The ITS sequences (Fig. 1) derived from MW882089 (Genbank number) i.e. the holotype for Russula pauriensis var. caulocystidii var. nov. was fixed among the other species of sub sect. Amoeninae and retrieved with 37% bootstrap value as a sister species to R. pauriensis (Genbank accession number: MF53185) from India. The tree shows the highest log likelihood (-3714.64). The extralimital taxa i.e. Cortinarius furvolaesus was chosen as an out group.
Russula laeta F.H. Møller & Jul. Schäff., Bull. Soc. Mycol. France: 231, t. 62 (1934)
Mycobank Number 305383 Fig. 7–12
Genbank Number MW 882091
Basidiocarp up to 6.7 cm in height. Pileus up to 5.8 cm in diameter, convex, umbo absent, surface brownish red (8C7) with purplish and pastel red (8A5) tinges; dry; glabrous; margins regular, splitting at maturity; cuticle half peeling ; flesh up to 0.4 cm broad, white (1A1), unchanging. Pileal veil absent. Odour fruity. Lamellae up to 0.5 cm broad, adnate, unequal, crowded, pale orange (6A3), unchanging; gill edges smooth. Stipe central, up to 6.1 cm long, 1.6 cm towards apex, 1.7 cm at centre and 2.2 cm at base, equal in diameter with distinctly bulbous base, white (1A1) with yellowish white (1A2) tinge at base; hollow; smooth; unchanging; exannulate.
Basidiospores 7.47–9.13 × 6.64–7.47µm; Q = 1.12–1.2 µm; globose to sub-globose; warty, warts 0.8–2.6 µm high, isolated, blunt, catenulate, incomplete reticulum; amyloid; apiculate, apiculus 1.6–3.2 µm long. Basidia 33.6–48.0 × 9.6–14.4 µm, clavate, granular, tetra-sterigmate, sterigmata 4.8-8.0 µm, granular, pointed. Pleurocystidia 52.0–70.4 × 8.0–12.8 µm, clavate, broadly clavate to sub-cylindrical, blunt and acute apices, granular. Cheilocystidia 40.0–54.4 × 8.0–12.8 µm, clavate with inflated to beaked tips, granular. Hymenophoral trama cellular. Gill edges sterile. Pileus cuticle gelatinized, gelatinization up to 85 cm broad, epicutis made up of 3.2–4.98 µm broad dermatocystidia, hairy, filamentous, loosely arranged, encrustated, granular, septate hyphae, context heteromerous, made up of 3.2–6.4 µm broad, septate, granular hyphae intermixed with 4.8–22.4 µm, irregular, granular cells. Stipe cuticle hyphal, gelatinized, made up of 2.4–6.4 µm broad, granular, septate hyphae intermixed with 3.2–8.0 µm, granular, irregular cells, giving rise to caulocystidia; context cellular, made up of 3.2–40.0 µm broad, hyaline, irregular cells forming spherocystis intermixed with 3.2–6.4 µm broad, septate, granular hyphae. Caulocystidia 38.4–56.0 × 6.4–11.2 µm, sub-cylindical to fusoid ventricose with blunt and acute apices, densely granular, septate. Clamp connections absent.
Collection examined Narkanda (2636m) 31̊ 15’6ʺN − 77̊ 28’20ʺE, growing solitary on moss in pure Cedrus deodara forest during late monsoon season, October 13, 2018, Amandeep Kaur, PUN 11186.
Results This collection falls very close to Russula laeta in its morpho-anatomical details as well as molecular analysis. Although according to [15] it seems to be very similar to R. velenoveskyi but the cap in this species does not have any purplish or copper tinges as are present in above described species and R. laeta. Also the size of the basidiospores and dermatocystidia of the present collection are in an arrangement with R. laeta. This species has been not earlier reported from India. So, it is the first report of this species from India. The molecular analysis was done for futher conformity which gives 100% query cover and 98.73% of percentage identification with R. laeta.
Phylogeny The sequences derived from our second species (Fig. 12), R. laeta (Genbank accession number: MW882091) is placed among the other species of subsect. Integrae and clustered with unidentified R. species (Genbank accession number: MK389376) from Pakistan showing 50% bootstrap value and formed a sister clade to R. laeta (Genbank accession number: MK386826 and MK389377). The tree displays the highest log likelihood (-2427.83). The extralimital taxa i.e. Russula rajendrae was chosen as an out group.
Russula amethystina Quѐl., Comptes Rendus de l' Association Franҫaise pour l' avancement des Sciences 26 (2): 450, 4:13 (1898)
Mycobank Number 174401 Fig. 13–16
Genbank MW882093
Basidiocarp up to 8.0 cm in height. Pileus up to 5.5 cm in diameter, convex; umbo absent; surface dark brown (6F5) with grayish brown (6D3) and pale yellow (1A3) tinges towards center; scales absent; margins regular, spilitting at maturity, dry; cuticle half peeling; flesh up to 0.4 cm thick, yellowish white (3A2), unchanging; odour mild. Pileal veil absent. Lamellae up to 0.8 cm, moderately broad, adnate, subdistant, equal, pale orange (5A3); gill edges smooth. Stipe central, up to 6.2 cm long, up to 1.5 cm at apex, up to 2.4 cm at center and 3.0 cm in diameter at base, obclavate; white (1A1); smooth; hollow; exannulate.
Basidiospores 8.0–11.2 × 6.4–9.6 µm; Q = 1.25–1.16 µm; sub-globose, warty, warts 0.8–1.6 µm, warts 2–3 connected, incomplete reticulum; amyloid; apiculate, apiculus 2.4–4.8 µm long, supra-hilar pledge present near apiculus. Basidia 36.8–60.8 × 9.6–17.6 µm, clavate to broadly clavate, granular, bi - tetrasterigmate, sterigmata up to 3.2–8.0 µm long, hyaline to granular, pointed. Pleurocystidia 54.4–89.6 × 11.2–16.0 µm, clavate to fusoid ventricose with blunt to beaked tips, granular, protruding out of the basidial layer and deeply placed. Cheilocystidia 32.0–48.0 × 6.4–9.6 µm, clavate with rounded and blunt tips, granular. Hymenophoral trama hyphal as well as cellular. Gill edges fertile, heteromerous. Pileus cuticle hyphal intermixed with pileocystidia, made up of 3.2–6.4 µm broad, highly septate, horizontally tangled hyphae, granular, intermixed with 3.2–4.8 µm broad, irregular, granular cells; context hyphal made up of 3.2–12.0 µm broad, septate, granular hyphae intermixed with 3.2–32.0 µm, irregular, hyaline to granular cells. Oleiferous hyphae are present. Pileocystidia 27.2–46.4 × 4.8–6.4 µm, clavate with rounded, blunt apices, granular. Stipe cuticle hyphal, made up of 3.2–6.4 µm, longitudinally tangled, granular, septate hyphae giving rise to caulocystidia intermixed with 3.2–8.0 µm broad, irregular, granular cells; context hyphal, made up of 3.2–8.0 µm broad, longitudinally tangled, septate, granular hyphae intermixed with 3.2–19.5 µm broad, irregular, hyaline to granular cells. Oleiferous hyphae are present. Caulocystidia 14.4–30.4 × 4.8–7.2 µm, clavate to fusoid ventricose with round, blunt apex, granular. Clamp connections absent.
Collection examined Hattu Peak road (2945 m) 31̊ 15’6ʺN- 77̊ 29’ 19ʺE, growing solitary on Cedrus deodara needles in mixed forest during late monsoon season, 13th October 2018, Amandeep Kaur, PUN 11183.
Edibility Edible, flesh eating insect was reported.
Results All the morphological and anatomical characters matches well with the details given for Russula amethystina [15, 16]. The present collection is almost similar to its closely allied R. turci Bres., but due to the presence of sub isolated warts forming incomplete reticulum and presence of yellow tinges on the pileus due to persistent rainfall it is placed under R. amethystine. [16] also differentiate the two allied species on the basis of odour which is weaker in R. amethysina whereas more clear and limited to the base of stem in R. turci. The present collection differs from those in possessing much larger spore size [15, 16]. Morpho-anatomical characters matches well with molecular characterization of Russula amethystina. Earlier this species is reported for the first time from India by [17]. Now, it is rereported from India.
Phylogeny The ITS sequences from R. amethystina (Genbank accession number: MW882093) (Fig. 16), are grouped with other species in the subsection Amethystinae and placed with an unidentified R. amethystina from Pakistan (Genbank accession number: KT953612), forming a sister clade to R. cuprea (Genbank accession number: KT953616) with an 75% bootstrap value. The tree displays the highest log likelihood (-4124.55). Lactifluus rajendrae was used as an extralimital taxon.
Russula globispora (J. Blum) Bon, Documents Mycologiques 17 (65): 55 (1986)
Mycobank Number 129550 Fig. 17–22
Genbank Number MW882090
Basidiocarp up to 5.2 cm in height. Pileus up to 4.8 cm in diameter, applanate; umbo absent; surface grayish orange (6B3) with brownish orange (6C5) tinges towards margins; scales absent; margins regular, spilitting at maturity, moist; cuticle half peeling; flesh up to 0.4 cm thick, white (1A1), unchanging; odour fragrant. Pileal veil absent. Lamellae up to 0.6 cm moderately broad, adnexed forked towards stipe, lamellulae rare, close, equal, orange white (6A2); gill edges smooth. Stipe excentric, up to 4.2 cm long, up to 1.8 cm at apex, up to 1.5 cm at center and 1.7 cm in diameter at base, obclavate; white (1A1) with pale orange tinges at base; smooth; solid; exannulate.
Basidiospores 8.0–12.8 × 7.2–10.4 µm; Q = 1.1 − 1.2 µm; sub-globose, warty, warts 0.8–1.6 µm, warts isolated, rarely connected, incomplete reticulum; amyloid; apiculate, apiculus 3.2–4.8 µm long. Basidia 24.0–60.8 × 9.6–16.0 µm, clavate to broadly clavate, hyaline to granular, bi - tetrasterigmate, sterigmata up to 0.8 − 6.4 µm long, hyaline, pointed. Pleurocystidia 72.0–97.6 × 10.4–17.6 µm, clavate to broadly clavate with blunt to beaked tips, granular, protruding out of the basidial layer and deeply placed, abundant. Cheilocystidia 35.2–49.6 × 8.8–12.8 µm, clavate to broadly clavate with rounded, beaked, blunt and capitate tips, granular. Hymenophoral trama hyphal as well as cellular. Gill edges fertile, heteromerous. Pileus cuticle hyphal intermixed with pileocystidia, geletinized, ixocutis, made up of 2.4–8.0 µm broad, highly septate, horizontally tangled, branched hyphae, granular, intermixed with 2.4–4.8 µm broad, irregular, granular cells; context hyphal made up of 3.2–8.0 µm broad, septate, granular hyphae intermixed with 4.8–41.6 µm, irregular, hyaline to granular cells. Oleiferous hyphae are present in cuticle as well as context. Pileocystidia 40.0–96.0 × 6.4–11.2 µm, clavate to broadly clavate to sub clavate with blunt apices, granular. Stipe cuticle hyphal, made up of 3.2–9.6 µm, longitudinally tangled, granular, septate hyphae giving rise to caulocystidia intermixed with 3.2–12.8 µm broad, irregular, granular cells; context hyphal, made up of 3.2–10.4 µm broad, longitudinally tangled, septate, granular hyphae intermixed with 4.0–25.6 µm broad, irregular, hyaline to granular cells. Caulocystidia 32.0–48.0 × 6.4–11.2 µm, clavate to fusoid ventricose with round, blunt apex, hyaline to granular. Clamp connections absent.
Collection examined Narkanda (2044 m) 31̊ 17’3ʺN- 77̊ 27’ 17ʺE, growing solitary on leaf litter in mixed forest during late monsoon season, 14th October 2018, Amandeep Kaur, PUN 11185.
Results All the morpho-anatomical characters matches well with the description given for Russula globispora [15, 18]. The present collection is featured by the presence of grayish orange (6B3) with brownish orange (6C5) tinges towards margins, lamellae forked towards stipe, basidiospores isolated to rarely connected (mostly 2 warts connected), presence of pileocystidia in pileus cuticle along with highly septate projecting hyphae with intermixed oleiferous hyphae described by [15]. Morpho-anatomical characters matches well with molecular characterization of Russula globispora. This species has been not earlier reported from India. So, it is the first report of this species from India. The molecular analysis was done for futher conformity which gives 95% query cover and 96.31% of percentage identification with R. globispora.
Phylogeny The sequences derived from our second species (Fig. 22), R. globispora (Genbank accession number: MW882090) is placed among the other species of subsect. Urentes and clustered with unidentified R. species (Genbank accession number: MG386702) from Slovakia showing 53% bootstrap value and formed a sister clade to uncultured Russula (Genbank accession number: LC367737). The tree displays the highest log likelihood (-1575.26). The extralimital taxa i.e. Cortinarius bubulus was chosen as an out group.
Russula cuprea Krombh., Naturgetreue Abbildungen und Beschreibungen der Schwӓmme 9:11, t. 66:1–3 (1845)
Mycobank Number 251691 Fig. 23–26
Genbank Number MW882092
Basidiocarp up to 5.2 cm in height. Pileus up to 4.7 cm in diameter, applanate; umbo absent; surface pale red (10B3) to violet brown (10E4) with light green tinges and pale yellow center; scales absent; margins regular, spilitting at maturity, striated; dry; cuticle fully peeling; flesh up to 0.3 cm thick, white (1A1), unchanging; odour mild. Pileal veil absent. Lamellae up to 0.6 cm broad, adnexed, crowded, equal, pale orange (5A2), interveined bottom; gill edges smooth. Stipe central, up to 3.9 cm long, up to 1.0 cm at apex, up to 1.1 cm at center and 1.3 cm in diameter at base, almost equal in diameter throughout with slightly bulbous base; white (1A1), turns yellowish on handling; smooth; hollow; exannulate.
Basidiospores 8.3–10.79 × 6.64–9.13 µm; Q = 1.25 − 1.18µm; globose to broadly ellipsoid, warty, warts 0.8–3.32 µm, warts isolated, incomplete reticulum, pladge present; amyloid; apiculate, apiculus 1.66–3.32 µm long. Basidia 41.6–56.0 × 12.8–17.6 µm, clavate to broadly clavate, densely granular, tetrasterigmate, sterigmata upto 5.6–9.6 µm long, granular, pointed. Pleurocystidia 62.4–92.8 × 9.6–14.4 µm, clavate with long tubular neck with captate to beaked to flamed tips, granular, protruding out of the basidial layer and deeply placed, abundant. Cheilocystidia 43.2–64.0 × 6.4–11.2 µm, clavate to subcylindrical, mucronate having inflated tips, granular. Hymenophoral trama cellular. Gill edges fertile, heteromerous. Pileus cuticle hyphal intermixed with pileocystidia, geletinized, ixocutis, made up of 3.2–6.4 µm broad, thin hairy, septate, horizontally tangled hyphae, granular, intermixed with 3.2–6.4 µm broad, irregular, granular cells, gelatinization up to 120 µm thick; context hyphal made up of 3.2–8.0 µm broad, septate, granular hyphae intermixed with 4.8–32.0 µm, irregular, hyaline to granular cells. Pileocystidia 51.2–97.6 × 8.0–12.8 µm, long cylindrical to obtuse to pyriform with blunt tips, granular. Stipe cuticle hyphal, made up of 2.4–4.8 µm, longitudinally tangled, granular, septate hyphae giving rise to caulocystidia intermixed with 4.0–8.0 µm broad, irregular, granular cells; context hyphal, made up of 3.2–8.0 µm broad, longitudinally tangled, septate, branched, granular hyphae intermixed with 4.0–33.6 µm broad, irregular, hyaline to granular cells. Caulocystidia 16.0–41.6 × 4.8–12.8 µm, clavate to fusoid ventricose with round, blunt apex, granular. Clamp connections absent.
Collection examined Narkanda (2619m) 31̊ 15’6ʺN- 77̊ 28’ 21ʺE, growing solitary near Cedrus deodar on humicolous soil in coniferous forest during late monsoon season, 13th October 2018, Amandeep Kaur, PUN 11184.
Results The morpho-anatomical characters goes well with the description given for Russula cuprea [15, 16]. The present collection is featured by the presence of pale red to violet brown surface with light green tinges and pale yellow center, stipe white turns yellow on handling, pileus having thin hairy turf of hyphae with embeded long cylindrical to obtuse blunt tiped elements which are similar in our species and are described as pileocysidia as described by Romagnesi. This species has been not earlier reported from India. So, it is the first report of this species from India. Morpho-anatomical characters matches well with molecular characterization of Russula cuprea. This species has been not earlier reported from India. Hence, this species is the first record from India. The molecular analysis was done for futher conformity which gives 95% query cover and 99.66% of percentage identification with R. cuprea.
Phylogeny The ITS sequences (Fig. 26), from R. cuprea (Genbank accession number: MW882092), are placed with the other species of the subsection Urentes and grouped with unidentified R. sp. from China (Genbank accession number: KX441137) and formed a sister clade to R. cuprea (Genbank accession number: KF850403) showing 82% bootstrap value. In the tree, the highest log likelihood is shown (-2013.85). Lactarius alboscrobiculatus, an extralimital taxon, was selected.