Research Article |
Corresponding author: Nakarin Suwannarach ( suwan.462@gmail.com ) Academic editor: Danushka Sandaruwan Tennakoon
© 2024 Wenhua Lu, Pumin Nutaratat, Jaturong Kumla, Saowaluck Tibpromma, Abdallah M. Elgorban, Samantha C. Karunarathna, Nakarin Suwannarach.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Lu W, Nutaratat P, Kumla J, Tibpromma S, Elgorban AM, Karunarathna SC, Suwannarach N (2024) Morphological and molecular identification of two new Marasmiellus species (Omphalotaceae, Agaricales) from Thailand. MycoKeys 109: 31-48. https://doi.org/10.3897/mycokeys.109.129791
|
Marasmiellus (Omphalotaceae, Agaricales) specimens collected in Thailand were investigated based on morphological characteristics and molecular phylogenetic analyses. In the present study, two species are introduced as new to science, namely Marasmiellus thailandicus and M. minutisporus. Phylogenetic analyses were carried out based on the internal transcribed spacer (nrITS) and nuclear ribosomal RNA large subunit (nrLSU) regions, and the results revealed that the two new taxa are distinct species within Marasmiellus. Another specimen was identified as M. scandens and is reported for the first time with morphology and molecular data from Thailand. Descriptions, illustrations, and phylogenetic results are provided. In addition, M. diaphanus and M. colocasiae are proposed as new combinations of Collybiopsis diaphana and Paramarasmius colocasiae, respectively, based on the phylogenetic evidence.
Basidiomycota, new taxa, omphalioid mushroom, taxonomy, wood-decaying mushroom
The genus Marasmiellus Murrill was proposed by
Some phylogenetic studies on Omphalotaceae based on nrITS and nrLSU in single and multigene analyses have been conducted to solve the stable placement where Gymnopus (Pers.) Gray and Marasmiellus still remain uncertain. Initially, Gymnopus and Marasmiellus were treated as multiple branches, nonmonophyletic groups (
According to the recent nomenclature, the type species of Marasmiellus (M. juniperinus Murrill) was transferred to Collybiopsis (J. Schröt.) Earle.
The present study aims to describe two new Marasmiellus species and a new report of M. scandens (Massee) Dennis & D.A. Reid collected from Thailand based on morphological characteristics and multigene phylogenetic analyses. In addition, we propose two new combinations of Collybiopsis diaphana and Paramarasmius colocasiae. Descriptions, illustrations, and a phylogenetic tree to show the placement of the taxa are provided.
Fresh basidiomata were collected from the northern part (Chiang Mai University, Chiang Mai Province) and the southern part (Phatthalung Province) of Thailand. Macromorphological features of the basidiomata were documented and photographed in the field. Color names and codes were determined following
The genomic DNA was extracted from fresh specimens using the DNA Extraction Mini Kit (FAVORGEN, China) according to the manufacturer’s instructions. Primer pairs ITS1/ITS4 (nrITS) and LR0R/LR5 (nrLSU) regions were amplified by the polymerase chain reaction (PCR) (
The newly generated forward and reverse of sequences from this study were assembled in the BioEdit v. 7.0.5 (
Names, voucher numbers, countries, and corresponding GenBank accession numbers of the taxa used in the phylogenetic analyses of this study.
Taxa Name | Voucher | Country | GenBank Accession Number | |
---|---|---|---|---|
nrITS | nrLSU | |||
Collybiopsis biformis | TENN58541 | USA | DQ450054 | NA |
C. biformis | HMJAU61116 | China | OQ597035 | OQ594445 |
C. brunneogracilis | SFSU-AWW01 | Indonesia | AY263434 | NA |
C. carneopallida | BRNM:747442 | Italy | OM522632 | NA |
C. clavicystidiata | SFC20180705–84 | South Korea | OL467252 | OL462817 |
C. confluens | HMJAU61120 | China | OQ597037 | NA |
C. confluens | TENN50524 | Sweden | DQ450044 | NA |
C. confluens | TENN-F-067864 | Germany | KP710296 | NA |
C. diaphana | Cesar202 | Mexico | MT232390 | NA |
C. diaphana | Cesar44 | Mexico | MT232391 | NA |
C. furtive | SFSU-F-024524h1 | USA | MN413341 | NA |
C. furtive | SFSU-F-024524h2 | USA | MN413342 | NA |
C. istanbulensis | KATO fungi 3596 | Turkey | KX184795 | KX184796 |
C. juniperina | TENN59540 | USA | AY256708 | KY019637 |
C. juniperina | TENN-F-58988 | Argentina | KY026661 | KY026661 |
C. luxurians | TENN-F-057910 | USA | AY256709 | AY256709 |
C. luxurians | HMJAU61101 | China | OQ597045 | OQ594455 |
C. luxurians | HMJAU61198 | China | OQ597046 | OQ594456 |
C. melanopus | SFSU AW54 | Indonesia | OR818034 | OR817634 |
C. melanopus | CUH AM093 | India | KM896875 | KP100305 |
C. orientisubnuda | NIBRFG0000500990 | Turkey | OL467262 | OL546546 |
C. peronata | TENN-F-065120 | Belgium | KY026677 | KY026677 |
C. quercophila | TENN-F-69267 | Slovakia | KY026729 | NA |
C. quercophila | TENN-F-69320 | USA | KY026736 | NA |
C. ramealis | TENN-F-065146 | Belgium | MN413346 | MW396882 |
C. ramealis | TENN-F-065145 | Belgium | MN413345 | MN413345 |
C. ramulicola | GDGM44256 | China | KU321529 | NA |
C. stenophylla | TENN-F-051099 | USA | MN413330 | MW396887 |
C. stenophylla | TENN-F-065943 | USA | MN413331 | MW396886 |
C. ugandensis | SFSU-BAP 614 | Sao Tome | MF100986 | NA |
C. vellerea | SFC20140821-29 | South Korea | OL467267 | OL462810 |
Gymnopus alkalivirens | TENN51249 | USA | DQ450000 | NA |
G. brunneiniger | XAL-Cesar 49 | Mexico | MT232389 | NG075396 |
G. efibulatus | HGASMF01-7052 | China | OM970865 | OM970865 |
G. fusipes | TENN59300 | China | AF505777 | NA |
G. fusipes | TENN59217 | France | AY256710 | AY256710 |
Marasmiellus agrianum | NJ201111 | Pakistan | MZ044839 | NA |
M. agrianum | NJ201112 | Pakistan | MZ044840 | NA |
M. alnicola | URM90019 | Brazil | KY302681 | KY302682 |
M. bicoloripes | CAL1524 | India | KY807129 | KY817233 |
M. candidus | CBS:252.39 | USA | MH856003 | NA |
M. candidus | MSM#0017 | Pakistan | KJ906507 | NA |
M. celebanticus | TO HG2281 | Spain | JF460781 | NA |
M. gregarius | G0197 | Japan | NA | MK278330 |
M. griseobrunneus | AMH 10117 | India | MK656132 | MK660195 |
M. griseobrunneus | CAL 1752 | India | MK660191 | MK660192 |
M. griseobrunneus | AMH 10118 | India | MK660194 | MK660193 |
M. lucidus | s1 | China | OP459424 | NA |
M. lucidus | HT10 | Japan | AB968237 | AB968237 |
M. minutisporus | CMUB40054 | Thailand | PP889931 | PP890011 |
M. minutisporus | CMUB40055 | Thailand | PP889932 | PP890012 |
M. omphaloides | PDD:95810 | New Zealand | HQ533031 | NA |
M. paspali | AHH65 | USA | EF175515 | NA |
M. paspali | AHH26 | USA | EF175511 | NA |
M. pilosus | iNat91483993 | Cayman Islands | OP651730 | NA |
M. rhizomorphogenus | BRNM:715003 | South Korea | GU319116 | GU319120 |
M. scandens | GH-80 | Ghana | MN794179 | NA |
M. scandens | GH-21 | Ghana | MN794139 | NA |
M. scandens | KUNCC22-12451 | China | OP536418 | NA |
M. scandens | CMUB40056 | Thailand | PP889933 | PP890013 |
M. thailandicus | CMUB40052 | Thailand | PP889929 | PP890009 |
M. thailandicus | CMUB40053 | Thailand | PP889930 | PP890010 |
M. tenerrimus | TENN61596H1 | USA | FJ596840 | NA |
M. tenerrimus | TENN61596H2 | USA | FJ596841 | NA |
M. tricolor | M01452 | Estonia | LR872638 | NA |
M. venosus | TNS-F-52281 | Japan | AB968236 | NA |
M. violaceogriseus | PDD:95788 | New Zealand | HQ533014 | NA |
M. volvatus | URM 84466 | Brazil | KC348449 | KC348442 |
M. sacchari | CBS:215.32 | USA | NA | MH866745 |
Moniliophthora perniciosa | CMR UB 2041 | Brazil | AY317136 | NA |
Paragymnopus foliiphilus | TENN-F-68183 | USA | KY026705 | KY026705 |
P. perforans | TENN-F-50318 | Sweden | KY026623 | KY026623 |
P. perforans | TENN-F-50319 | Sweden | KY026624 | KY026624 |
P. pinophilus | TENN-F-69207 | USA | KY026725 | KY026725 |
Paramarasmius colocasiae | SP376044 | Brazil | GQ452780 | NA |
Pa. mesosporus | TNS-T-48339 | Japan | OM522625 | OM522623 |
Pa. palmivorus | AKD 112/2015 | India | MG251431 | MG251441 |
Paramycetinis austrobrevipes | TENN-F-50135 | Australia | KY026622 | KY026622 |
Par. caulocystidiatus | TENN-F-5405 | New Zealand | KY026645 | NA |
Pseudomarasmius efibulatus | TENN-F-56187 | New Zealand | MK268234 | NA |
Ps. glabrocystidiatus | BRNM 718676 | Korea | NR152899 | KF251093 |
Ps. nidus-avis | Cesar36 | Mexico | MH560576 | NA |
Ps. pallidocephalus | TENN-F-52401 | USA | KY026635 | KY026635 |
Ps. patagonianus | TENN-F-54424 | Chile | KY352649 | NA |
Ps. quercophylloides | TENN-F-49177 | China | MK268235 | NA |
Pusillomyces manuripioides | JO674 | Brazil | MK434210 | MK434211 |
Pu. manuripioides | JO1121 | Brazil | MK434212 | MK434213 |
In the dataset, the combined nrITS and nrLSU sequence dataset consisted of a total number of 86 taxa, and the aligned dataset was comprised of 1593 characters, including gaps (nrITS: 1–709 and nrLSU: 710–1593). The best RAxML tree was obtained with a final ML optimization likelihood value of -15536.515830. The matrix had 780 distinct alignment patterns, with 34.45% undetermined characters or gaps. Estimated base frequencies were as follows: A = 0.248165, C = 0.187111, G = 0.251637, T = 0.313087, with substitution rates AC = 1.027131, AG = 5.242142, AT = 1.916708, CG = 0.743477, CT = 6.311829, GT = 1.000000; gamma distribution shape parameter α = 0.681313, tree-Length = 3.382466. Notably, the phylograms of the ML and BI analyses were similar in topology. Therefore, the phylogenetic tree obtained from ML analysis was selected and is presented in this study (Fig.
A combined phylogenetic tree was generated from maximum likelihood analysis (RAxML) based on a combined nrITS and nrLSU dataset. Bootstrap values (BS) ≥ 60% from ML analysis (left) and Bayesian posterior probabilities (right) (PP) ≥ 0.90 are shown on the branches. Newly sequenced collections are indicated in red, new combinations are in blue, and the type specimens are denoted by ‘T’.
Two specimens in this study, CMUB40054, and CMUB40055, were clustered in a well-supported lineage (1.00 PP/100% BS), introduced as M. minutisporus related to M. pilosus (Dennis) Singer (iNat91483993), and sister to M. griseobrunneus Sharafudheen & Manim (AMH 10117, AMH 10118, and CAL1752) with a support value of 0.99 PP and 84% BS). The second species of this study (CMUB40052 and CMUB40053) formed a separate lineage within the genus Marasmiellus with higher support values (1.00 PP/100% BS) and formed a clade with M. bicoloripes K.P.D. Latha, K.N.A. Raj & Manim (CAL1524) and M. minutisporus cluster, and we introduced it as M. thailandicus. Additionally, one collection (CMUB40056) was clustered with M. scandens strain (GH80 and GH21) with strong statistical support (0.99 PP/100% BS). Noteworthily, M. scandens (KUNCC22-12451) differs from M. scandens (GH21) and (GH80) by 29 base pair differences out of 577, but our strain (CMUB40056) only has two base pair differences out of 570 bases (Fig.
“thailandicus” refers to the country Thailand, where the type species was collected.
Differs from M. candidus by the presence of a reddish gray to dull red, dry, radially wrinkled surface, grooved pileus with distant lamellae, and obovate or ellipsoid spores.
Basidiomata small-sized, marasmioid. Pileus 6–20 mm diam., hemispherical first, expanding to plano-convex with slightly concave, radially sulcate with age, with involute then deflexed or straight margin, not distinctly hygrophanous, translucently striate up to center, reddish gray (10B2) to dull red (10B3) all over with grayish red (7B3) center, grayish red (7B3) margin, surface smooth, dry, and dull. Lamellae distant, broadly adnate to subdecurrent, sometimes anastomosing, white to cream-colored, with a reddish white (7A2) edge, particularly in young specimens, I = 2–3, L = 11–14. Stipe 4–8 × 1–2 mm, cylindrical, curved, base often slightly swollen, inconspicuous fibrils or scurfy, creamy-white at apex, slightly reddish white (7A2) at the base, dry, pruinose all over, Context thin, fistulose or solid, concolorous with surface. Smell and taste indistinct.
Basidiospores (–12)13–16(–17) × 4–5(–6) μm (average = 15 × 5 μm), Q = (2.1)2.5–3.5(–3.75), Qm = 3.05 ± 0.38, sub-cylindrical to elongate with apiculus, inamyloid, thin-walled. Basidia 20–45 × 7–11 μm, 4-spored, clavate, sterigmata up to 5 μm. Cheilocystidia 50–53 × 8–13 μm, clavate. Pleurocystidia absent. Trama hyphae cylindrical, thin-walled, hyaline, inamyloid. Pileipellis, a cutis with transitions to a trichoderm, made up of cylindrical; pigment brown, intracellular and minutely incrusting, hyphal 2–4 μm, negative in Melzer’s reagent. Stipitipellis, a cutis of parallel, somewhat skewed, cylindrical, or clavate-shaped, smooth hyphae, thin-walled, up to 7.5 um wide. Caulocystidia absent. Clamp connection present.
Caespitose, in small groups growing on the bark of Lagerstroemia macrocarpa. Known only from the type locality in northern Thailand.
Thailand • Phatthalung Province, Khuan Khanun, Sago Palm (Metroxylon sagu) Forest, 7°44'02"N, 99°59'47"E; elevation 23 m; on decaying leaf and branches of deciduous tree; 6 September 2023; P. Nutaratat, P., Suwannarach & J. Kumla, (CMUB40054). GenBank accession numbers PP889930 (nrITS) and PP890011 (nrLSU).
“minutisporus” refers to the small basidiospores of this species.
Differs from M. virgatocutis by the grayish-brown, convex, wrinkled pileus, longer pileus terminal, smaller elongated spores, and caulocystidia.
Basidiomata small-sized, marasmioid. Pileus 5–11 mm diam., thin, then expanding to applanate, with slightly inflexed, pulvinate when young, then deflexed, finally reflexed, and undulating margin, convex when age with depressed to umbilicate at disc., grayish brown (7D3), often with grey, brown, or dark gray (1F1) at the center, gray at the margin; often radially wrinkled, surface dry, slightly pruinose to tomentose under the lens. Lamellae distant, often more or less reduced, white to sordid beige with a concolorous overall, pruinose edge, I = 1–3, L = 13–16. Stipe 4–9 × 1 mm, cylindrical, often subbulbous at the base, off-white at the apex, fourth to fifth downward, and black or gray at the stipe base, entirely white pruinose, with basal tomentum. Context thin, soft, white, fistulose. Smell and taste none.
Basidiospores (11)12–15(–16) × (3)4–5 μm (average = 14 × 4 μm), Q = (2.6)2.8–4(4.67), Qm = 3.5 ± 0.56, hyaline, inamyloid, cylindrical to fusiform, thin-walled. Basidia 25–28 × 7–9 μm, 4-spored, sterigmata up to 5 μm long, clavate. Lamella edge sterile. Cheilocystidia 30–33 × 12–15 μm, somewhat clavate to subglobose, Pleurocystidia absent. Pileipellis a cutis made up of 4.0–10 μm wide, inamyloid, inflated or cylindrical hyphae, with scattered suberect to erect, thin-walled, smooth, sometimes terminal elements up to 61–99 × 3.0–10 μm, gradually tapering to an acute or rounded apex. Stipitipellis a cutis composed of cylindrical, inamyloid, parallel, slightly, not incrusted, smooth, thick-walled. Caulocystidia 25–50 × 4–5 μm, adpressed to erect, cylindrical, clavate, smooth, thin-walled. Clamp connection present.
Solitary to caespitose, in small groups growing on decaying leaves and twigs of deciduous trees. Known only from the type locality in southern Thailand.
Thailand • Phatthalung Province, Khuan Khanun, Sago Palm (Metroxylon sagu) Forest, 7°44'02"N, 99°59'46"E; elevation 23 m; on decaying leaf and twigs of deciduous tree; 7 September 2023; P. Nutaratat, N. Suwannarach & J. Kumla, (CMUB40055). GenBank accession numbers PP889932 (nrITS) and PP890012 (nrLSU).
Basidiomata small-sized, marasmioid. Pileus 5–10 mm diam., orbicular when young and then convex at age, streaked from disc to margin, margin entire, wavy to irregular, decurved or greatly reflexed, surface dry, smooth, white pruinose, white to grayish orange (5B4–5), grayish orange (5B4) at the margin. Lamellae adnate, subdistant, with 2–3 series of lamellulae, 14–16 major lamellae, unequal, narrow, pale white at face and edge. Smell and taste none. Stipe 4–6 × 1–2 mm, often curved, lateral or central, disc at the base, dry, surface smooth, whitish to pale orange (5A3); Context thick, fistulose, orange white (5A2–3).
Basidiospores 7–9(–10) × (3)3.5–4.5(–6) µm (average = 8 × 4.5 μm), Q = 1.4–1.8(–2), Qm = 1.74 ± 0.11, broadly ellipsoid, smooth, hyaline, inamyloid, thin-walled. Basidia 10–12.5 × 3–4.5 µm, clavate, 4-spored, sterigmata up to 1.5 µm. Cheilocystidia abundant, 15–17(–22) × 8–9(–11) µm, broom-cell type with finger-like excrescences in the upper half, hyaline, thin-walled. Pleurocystidia absent. Pileipellis with poorly Ramealis-structure, trama hyphae 4–9 µm, negative in Melzer’s reagent. Stipitipellis hyphae up to 5 μm wide, smooth, thin-walled, trama not observed, negative in Melzer’s reagent. Clamp connections present.
Caespitose, in small groups on decaying leaf of Metroxylon sagu in southern Thailand (This study), on Aquilaria sinensis (agarwood) trees in China (
≡ Marasmiellus diaphanus César, Bandala & Montoya, in César, Montoya, Bandala & Ramos, Mycol. Progr. 19(10): 1022 (2020)
≡ Marasmiellus colocasiae Capelari & Antonín, Cryptog. Mycol. 31(2): 138 (2010)
A combination of morphological characteristics and molecular phylogeny inferred from sequence data of the nrITS and nrLSU region revealed two new species of Marasmiellus (M. thailandicus and M. minutisporus) and one known species (M. scandens) in this study. According to the phylogeny results, Marasmiellus is well-clustered in a strongly supported clade (1.00 PP/100% BS) (Fig.
Morphologically, M. thailandicus is closely related to M. subnigricans (Murrill) Singer, M. candidus, and M. bicoloripes. Marasmiellus subnigricans (11−45 mm diam.) and M. candidus (3–22 mm diam) contain dingy cream and purely white to sordid white (
Based on morphological characteristics, M. minutisporus is closely related to M. virgatocutis Robich, Esteve-Rav. & G. Moreno and M. griseobrunneus. However, M. virgatocutis is distinguished by the radially fibrillose-virgate pileus surface and the variable shape of cheilocystidia, which range from clavate to lageniform to molariform. While M. griseobrunneus has a surface of dark brown, the largest pileus (8−38 mm diam.), and observed pleurocystidia compared with M. minutisporus (
Marasmiellus scandens was previously reported worldwide (in Asia, Africa, Oceania, and North America) as a pathogen and endophytic form in cocoa and coffee (
Prior to the present study, eight species of Marasmiellus have been reported based on morphological characteristics in Thailand viz. M. albofuscus, M. alliiodorus, M. amygdalosporus, M. candidus, M. chamaecyparidis, M. collybioides, M. corticum, and M. paspali (
The authors thank Shaun Pennycook for the nomenclatural advice and Russell Kirk Hollis for his help with the English correction.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This work was supported by Chiang Mai University and National Higher Education, Science Research and Innovation Policy Council, Thaksin University Research Grant Fiscal Year 2023, Thailand. Nakarin Suwannarach and Samantha C. Karunarathna thank CMU Visiting Researcher 2024, Chiang Mai University (Grant number CMUVP038/2567), Thailand. Samantha C. Karunarathna and Saowaluck Tibpromma thank the National Natural Science Foundation of China (No. 32260004), Yunnan Revitalization Talents Support Plan (High-End Foreign Experts and Young Talents Programs), and the Key Laboratory of Yunnan Provincial Department of Education of the Deep-Time Evolution on Biodiversity from the Origin of the Pearl River. The authors extend their appreciation to the Deputyship for Research and Innovation, "Ministry of Education" in Saudi Arabia for funding this research (IFKSUOR3-299-19).
Conceptualization: NS, SCK. Data curation: WL, JK, PN. Investigation: WL, NS, JK, PN, ST, SCK. Project administration: NS, SCK. Software: WL, JK. Supervision: NS, SCK. Writing- review and editing: WL, NS, JK, PN, ST, AME, SCK.
Wenhua Lu https://orcid.org/0000-0001-7283-7596
Pumin Nutaratat https://orcid.org/0000-0001-7924-4822
Jaturong Kumla https://orcid.org/0000-0002-3673-6541
Saowaluck Tibpromma https://orcid.org/0000-0002-4706-6547
Abdallah M. Elgorban https://orcid.org/0000-0003-3664-7853
Samantha C. Karunarathna https://orcid.org/0000-0001-7080-0781
Nakarin Suwannarach https://orcid.org/0000-0002-2653-1913
All of the data that support the findings of this study are available in the main text