FIRST RECORD OF INOSPERMA SUBSPHAEROSPORUM (INOCYBACEAE) IN INDONESIA Текст научной статьи по специальности «Биологические науки»

Turczaninowia 26, 2: 103-113 (2023) DOI: 10.14258/turczaninowia.26.2.8 http://turczaninowia.asu.ru

ШШ ISSN 1560-7259 (print edition)

M TURCZANINOWIA

¡¡j| ISSN 1560-7267 (online edition)

УДК 582.287:581.95(594) First record of Inosperma subsphaerosporum (Inocybaceae) in Indonesia

I. P. Putra1*, R. Riffiani2' 4, O. D. Nurhayat2' 5, A. I. Toemon3' 6

1IPB University, Darmaga Campus, Raya Dramaga St., Bogor, West Java, 16680, Indonesia.

E-mail: ivanpermanaputra@apps.ipb.ac.id; ORCID iD: https://orcid.org/0000-0001-8328-8764

2 Research Center for Applied Microbiology, National Research and Innovation Agency (BRIN), Raya Jakarta-Bogor St., No. Km. 46, Bogor, West Java, 16680, Indonesia

3 Palangka Raya University, Yos Sudarso St., Central Kalimantan, Indonesia 4E-mail: riniriffiani@gmail.com; ORCID iD: https://orcid.org/0000-0002-0783-0483 5E-mail: oktandn25@gmail.com; ORCID iD: https://orcid.org/0000-0002-2790-8743 6E-mail: agnestoemon@gmail.com; ORCID iD: https://orcid.org/0000-0002-4067-2931

* Corresponding author

Keywords: Basidiomycota, Borneo, morphology, phylogeny, wild mushroom.

Summary. Inosperma (Inocybaceae) was originally described as a subgenus of Inocybe. Some species of Inosperma are considered edible mushrooms, but this genus also includes a few toxic species with high levels of muscarine. To date, the knowledge of this genus in Indonesia remains poor. During a mushroom hunting in Central Kalimantan (Indonesia) by the Indonesian mushroom hunter community, some fruiting bodies of inocyboid fungi which consumed and traded by the local people were collected. At glance, the specimens resembled Inosperma or Pseudosperma by the mac-romorphological characters in the field. Our goal was to justify the taxonomical position of our inocyboid specimens based on morphological and molecular evidence. The fresh fruiting bodies were used for the analysis. The basidiomata was described based on the macro- and micromorphological characters. The molecular analysis and phylogenetic tree construction were performed based on ITS 1-2 sequences. The combination of morphological and molecular analyses confirmed our collection as Inosperma subsphaerosporum. Morphologically, our specimens can be determined by the conical applanate cap with umbo, incurved to decurved margin, dry pileus with fibrillose to rimulose surface, lamellae adnexed to sinuate, cylindrical stipe, subglobose to globose smooth basidiospores, and abundant cheilocystidia in clusters. The yellow intracellular contents of cheilocystidia distinguished our specimen from Pseudosperma, while the outline of basidiospores differed our specimens to closely morphological species of I. carnosibulbosum, I. akirnum, and I. saragum. The BLAST result revealed that our specimen exhibits 96 % similarity to I. subsphaerosporum as the top hit. The phylogenetic tree (RAxML) nested our specimens in the I. subsphaerosporum clade from type material. The current study provides the first information on the occurrence of I. subsphaerosporum in Indonesia. Future study should be done to analyze both nutritional and muscarine content of this Indonesian Inosperma.

Первая находка Inosperma subsphaerosporum (Inocybaceae) в Индонезии

И. П. Путра1*, Р. Риффиани2, О. Д. Нурхайят2, А. И. Тоемон3

1 Богорский институт сельского хозяйства, кампус Дармага, ул. Райя Дарамага, г. Богор, Западная Ява, 16680, Индонезия

2 Исследовательский центр прикладной микробиологии, Национальное агентство исследований и инноваций, ул. Райя Джакарта-Богор, д. 46, г. Богор, Западная Ява, 16680, Индонезия

3 Палангкарайский университет, ул. Йос Сударсо, г. Паланкарая, Центральный Калимантан, Индонезия

* Автор для переписки

Поступило в редакцию 19.01.2023 Принято к публикации 12.07.2023

Submitted 19.01.2023 Accepted 12.07.2023

Ключевые слова: Борнео, дикорастущие грибы, морфология, филогенез, Basidiomycota.

Аннотация. Inosperma - род грибов семейства Inocybaceae, первоначально описанный в качестве подрода рода Inocybe. Некоторые виды рода Inosperma считаются съедобными грибами, но этот род также включает несколько токсичных видов с высоким содержанием мускарина. На сегодняшний день изученность этого рода в Индонезии остается фрагментарной. Во время сбора грибов в Центральном Калимантане (Индонезия) индонезийским обществом грибников-любителей было собрано несколько плодовых тел иноцибоидных грибов, которые местные жители употребляли в пищу и продавали. По макроморфологическим признакам в поле образцы напоминали Inosperma или Pseudosperma. Нашей целью было установить видовую принадлежность собранных иноцибоидных образцов на основе морфологических и молекулярных данных. Для анализа использовали свежие плодовые тела (базидиомы). Базидиомы описаны по макро- и микроморфологическим признакам. Молекулярный анализ и построение филогенетического дерева проводили на основе последовательностей ITS 1-2. Сочетание морфологического и молекулярного анализов подтвердило нашу находку как Inosperma subsphaerosporum. Морфологически наши образцы можно определить по конической уплощенной шляпке с вершиной, вогнутой к неизогнутому краю, сухим ворсинкам с волокнистой или слегка ребристой поверхностью, пластинкам, прикрепленным выемкой, цилиндрической ножке, гладким полушаровидным до шаровидных базидиоспорам и многочисленным скоплениям хейлоцистид. Желтое внутриклеточное содержимое хейлоцистид отличало наш образец от Pseudosperma, а очертание базидиоспор отличало наши образцы от близких по морфологии видов I. carnosibulbosum, I. akirnum и I. saragum. Результат BLAST показал, что наш образец имеет 96%-е сходство с I. subsphaerosporum. Филогенетический анализ (RAxML) подтвердил принадлежность наших образцов к кладе I. subsphaerosporum из типового материала. Настоящее сообщение представляет информацию о первой находке I. subsphaerosporum в Индонезии. Для анализа содержания питательных веществ и мускарина в этой индонезийской иносперме необходимы дополнительные исследования.

Introduction

The genus Inosperma (Kühner) Matheny et Esteve-Rav. (typified by I. calamistratum) was initially conceived as a subgenus of Inocybe by Kühner (1980). Currently, Inosperma is recognized as a genus rank in Inocybaceae Jülich (Matheny et al., 2019). Members of this genus are characterized by small to medium-fruiting bodies (basidiomata), rimose to scaly cap, reddening context when bruised, hyaline basidia, phaseoliform basidiospores, thin-walled cheilocystidia, lack of pleurocystidia, and often with distinctive odors (Matheny et al., 2019). The genus Inosperma accommodates species which were previously distributed to Inocybe sect. Cervicolores Singer (1975), Maculata clade (Larsson et al., 2009), and Old-World tropical clades (Kropp et al., 2013; Pradeep et al., 2016). A total of 80 taxa have been recorded in this genus worldwide (Index Fungorum, 2023). Currently, Inosperma is comprised of about 55 species in Africa, Australasia, Asia, Europe, and North America (Matheny et al., 2019). The number will increase as several recently described species of Inosperma have been reported (A'ignon et al., 2021; Deng et al., 2021; Li et al., 2022), including I. subsphaerosporum.

Inosperma subsphaerosporum, a new species of Inosperma was recently reported from Hainan Tropical Rainforest National Park China (Deng et al., 2021). The specific epithet "subsphaerosporum" refers to the shape of its basidiospores. Morphologi-

cally, this species can be determined by the brown to yellowish, scaly-fibrillose cap, grayish to brownish dense lamellae, a cylindrical longitudinally fibrillose stipe with recurved fibrils at the apex, and a salty or musty smell (Deng et al., 2021). To date, the distribution of this species was only known from the type locality in Hainan Province (China), grow on soil and associated with Fagaceae (Deng et al., 2021). Previously, I. subsphaerosporum was phylogenetical-ly placed in an Old-World tropical clade, in which all members are described from South Asia. However, knowledge of this species in Southeast Asia has yet to be found.

The information on wild Indonesian inocyboid mushroom is scarce, and species of Inosperma are poorly known. No information provided regarding the I. subsphaerosporum strain history, nor herbarium collection number that deposited in Indonesia. In 2022 and 2023, our field examination on the distribution of Inocybaceae in Central Kalimantan (Borneo) in Indonesia has revealed the consumption of wild inocyboid mushroom by the local community. The foraging and consuming of wild inocyboid mushrooms have been done for decades. During the fungus foray held by the Indonesian mushroom hunter community (KPJI), we obtained some basidiomata of wild edible inocyboid mushroom. The goal of this study was aimed to reveal the taxonomical position of our inocyboid specimens based on morphological and molecular evidence.

Material and Methods

The specimens were obtained at Buntok, Dusun Selatan, Barito Selatan, Central Kalimantan, Indonesia (1°42'30.3"S, 114°49'25.8"E), in 2022 and 2023 during a mushroom hunting carried by the KPJI. The fruiting bodies were photographed in situ and ecological information (coordinate, substrate, vegetations) were noted. Some of the specimens were deposited to Herbarium Bandungense Indonesia with the collection number FIPIA-DEP59. The morphological characteristics were examined from fresh basidiomata in situ and in Mycology Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Indonesia and in the Integrated Laboratory of Bioproducts (iLaB), BRIN, Bogor, Indonesia. The macromorphological features including color, size, pileus and stipe ornamentation, margin, and lamellae were observed directly and by a loupe. The micromorphological characters of basidium, cystidia, spores (shape, size, color, ornamentation), and clamp connection were observed using light microscope. Specimens were also subjected to examination via scanning electron microscopy (SEM), which was prepared following the methods of Goldstein et al. (1992). Lamellae were cut into small pieces (5 x 5 mm), pre-fixed in 2.5 % glutaraldehyde of a cacodylate buffer with a pH of 8.4 at 27 °C for two days. Next, they were pre-fixed in 2 % tannic acid for six hours and washed with four different cacodylate buffers. The samples were dehydrated in 50-100 % ethanol series, infiltrated with t-butanol twice for 10 minutes, and freeze dried. Freeze dried samples were mounted on an aluminum stub with double-sided carbon tape and coated with gold. Samples observed using the JSM IT 200 SEM system (JEOL, Tokyo, Japan). The specimens were identified using related identification references (Matheny et al., 2019; Deng et al., 2021; Li et al., 2022).

Fresh basidiomata were used to isolate DNA. DNA was extracted using a Qiagen DNeasy Plant Mini Kit according to the manufacturer's instructions. A Thermo Scientific Arktik Thermal Cycler was used for DNA amplification (ThermoFisher-Scientific). The amplification reaction was carried out using the primer pairs ITS 5 (5'-GGA AGT AAA AGT CGT AAC AAG G-3') and ITS 4 (5'-TCC TCC GCT TAT TGA TAT GC-3') (White et al., 1990). The PCR amplification was performed in 50 |L total reaction containing 9 |L ddH2O, 1.5 |L of 10 pmol of each primer, 25 |L PCR mix from 2x PCR buffer for KOD FX Neo (Toyobo), and 2 |L 100 ng template DNA. The PCR condition was set as follows: initial denaturation at 94 °C for 5 minutes, followed by 30 cycles of denaturation at 94 °C for 30 seconds, annealing at 56 °C for 45 seconds, and extension at 72 °C for 1 minute. The final extension was set at 72 °C for 10 minutes. PCR product was analyzed using 1.5 % agarose gel electrophoresis run with TAE buffer (40 mm Tris-acetate, pH 8.0 1 mM EDTA), stained with FloroSafe DNA stain and visualized by the Gel Doc EZ Gel Documentation System (Biorad). PCR products were sent to the 1st Base Malaysia for sequencing.

The sequences were assembled using Chro-masPro software. The final aligned sequences were deposited in GenBank (https://www.ncbi.nlm.nih. gov/) to obtain the accession number. The sequence was subjected to Basic Local Alignment Search Tool (BLAST) in NCBI to compare the homology with previous data. Selected published sequences based on BLAST results (Table) were used for phyloge-netic tree analyses with Auritella hispida as the out-group. The phylogenetic tree of Randomized Axeler-ated Maximum Likelihood (RaxML) Black Box was generated on CIPRES (Stamatakis, 2014). All trees were then edited using TreeGraph Software version 2.9.2-622 beta. The Bootstrap value (BS) > 70 % was shown on the branch of the phylogenetic tree.

Table

Species, outgroup, herbarium voucher, and GenBank accession numbers used in this study

Species Collection Code ITS Accession Number

Auritella hispida Strain TH1009 KT378203

Inosperma aff. calamistratum Strain REH8420 JQ801390

I. aff. calamistratum DED8134 GQ892983

I. aff. fastigiellum Strain PBM3325 JQ801399

I. aff. latericium Strain TR109-02 JQ801405

I. aff. maculatum Isolate AFTOL-ID-476 DQ241778

I. africanum Voucher MR00387 MN096189

I. africanum Voucher HLA0383 MT534298

I. akirnum Voucher CAL1358 KY440085

Table (continued)

Species Collection Code ITS Accession Number

I. apiosmotum Strain PBM3020 JQ801385

I. bicoloratum Voucher ZT12187 GQ892984

I. bulbomarginatum Voucher HLA0417 MT534300

I. bulbomarginatum Voucher MR00357 MN096190

I. calamistratoides Strain PBM3384 JQ801393

I. calamistratum Strain PBM1105 JQ801386

I. carnosibulbosum TBGT12047 KT329448

I. cervicolor Voucher TURA-4761 JQ801395

I. cf. lanatodiiscum Voucher TURA1812 JQ408763

I. changbaiense Voucher HMJAU25861 MH047251

I. cyanotrichium Strain I37 JQ801396

I. dodonae Voucher STU-SMNS-STU-F-0901253 MW647615

I. flavobrunneum Voucher HLA0372 MT534290

I. gregarium Voucher CAL1309 KX852305

I. hainanense Voucher Zeng4937 MZ374070

I. hainanense Voucher Zeng4936 MZ374069

I. ismeneanum Voucher STU-SMNS-STU-F-0901561 MW647625

I. latericium Voucher PDD-92382 GU233367

I. misakaense Strain PC96234 JQ801409

I. monastichum Voucher STU-SMNS-STU-F-0901533 MW647631

I. mucidiolens Voucher WTU: DG1824 HQ201339

I. muscarium Voucher Zeng4720 MZ373978

I. muscarium Voucher Zeng4736 MZ373979

I. neobrunnescens Strain SAT0427406 JQ801411

I. rimosoides Isolate AFT0L-ID-520 DQ404391

I. rubricosum Strain PBM3784 KP308817

I. saragum Voucher CAL1360 KY440103

I. subsphaerosporum Voucher FHMU3154 MW403826

I. subsphaerosporum Voucher FHMU3153 MW403825

I. subsphaerosporum Voucher FIPIA-DEP59 OP880257

I. subsphaerosporum Voucher FHMU3155 MW403827

I. vinaceum Voucher AMB-18747 MW561108

I. viridipes Voucher HO-562693 KP641646

I. virosum TBGT753 KT329452

Results Taxonomy

Inosperma subsphaerosporum Y. G. Fan, L. S. Deng, W. J. Yu et L. Y. Liu, 2021, in Deng, Yu, Zeng, Liu and Fan, Phytotaxa 502(2): 172. (Figs 1-5)

Fruiting bodies medium in size. Cap 4-7 mm, conical in young basidiomata, plano-convex to ap-planate in mature stages with an obtuse umbo, margin incurved at first and then slightly decurved in time, surface moist to dry, pileus radially fibrillose to rimulose with patches, scales of veil remnants, margin rough to split in in time, yellowish brown to brown around umbo, yellowish white with paler col-

or compared to the central region towards the margin, golden yellow in very mature stages. Lamellae adnexed to sinuate, dense, with series of lamellula, 20-40 mm length, brownish to white, edge slightly serrate. Stipe 40-100 x 8-11 mm, cylindrical, hollow, equal from apex to base with a slightly swollen base, squamulose at the apex, fibrillose downwards, white to cream. Context solid, fleshy, white to cream. Basidiospores subglobose to globose, 5-7x 4-6 ^m, smooth, yellowish to tan, with a prominent lipid body and an indentation in the center region when observed using SEM. Basidia clavate, notable oil inclusion, 26-31 x 7-8 ^m in dimension, hyaline, 4-spored, sterigmata 3-4.5 ^m in length. Pleurocys-tidia absent. Cheilocystidia abundant, 21-41 x 8-11

^m, in clusters, mostly club-shaped, septate, primarily thin-walled and some with thick-walled, hyaline, with yellow intracellular cytoplasm. Hyphal trama composed of cylindric to inflated hyphae, 12-15 ^m in diameter, hyaline. Pileipellis is a cutis, golden yellow, with thin-walled, cylindrical hyphae, 7-10 ^m

in diameter. Oleiferous hyphae prominent, 3-4 ^m wide, pale to yellow brown, smooth, occasionally dichotomously branched. Hymenial and stipe trama with dense interwoven hyphae, thin-walled, yellowish, 5-10 ^m wide. Clamp connections are present in all tissues.

Fig. 1. Field photograph of Inosperma subsphaerosporum FIPIA-DEP59: A, B - upper side of pileus; C - lamellae. Bars = 5 cm.

Habitat: gregarious near surrounding Euphor-biaceae and Arecaceae trees.

Distribution: To date, known only from Hainan Province, China.

Specimen examined: "Buntok, Dusun Selatan, Barito Selatan, Central Kalimantan, Indonesia, 1°42'30.3"S, 114°49'25.8"E, 12 m. a. s. l., on soil mixed with leaves litter, XII 2022, V 2023, collected by Andi B and Agnes I Toemon, FIPIA-DEP59".

Molecular Analyses

The aligned nucleotide sequence was deposited to GenBank with the accession number ITS OP880257.

The BLAST result revealed that our specimens had high similarity to Inosperma subsphaerosporum from type material (96 %) as the top hits. The phylogenetic tree (Fig. 6) constructed from ITS sequences resolved specimen FIPIA-DEP59 was in the same clade of I. subsphaerosporum from type material with 93 % BS value. The phylogenetic tree displayed that I. subsphaerosporum FIPIA-DEP59 was a sister clade to I. flavobrunneum. Inosperma subsphaerosporum FIPIA-DEP59 was in the different clade to morphologically similar species I. carnosibulbosum, I. akirnum, and I. saragum.

Fig. 2. Field collection and consumption of Inosperma subsphaerosporum FIPIA-DEP59: A, B - fresh basidiomata collection of I. subsphaerosporum from field; C - consumption of I. subsphaerosporum by the local people. Bars = 5 cm.

Discussion

Currently, there are only eighty taxa of Inosperma (Inocybaceae) documented worldwide (Index Fun-gorum, 2023). Inosperma sphaerobulbosum was recently introduced as a new species from China by Deng et al. (2021). The current study reports for the first time the distribution of this species aside from the type locality (Hainan Province, China) and contributes a new information to Indonesian macrofun-gi. Our work revealed the extension of geographical distribution of I. sphaerobulbosum in Kalimantan (Indonesia), which approximately 2474 km away from the type locality. Previously, Putra and Hafazal-lah (2020) reported the unidentified edible mushroom from Central Java (Indonesia) with the local name "jamur lengkuas" morphologically similar to I. sphaerobulbosum FIPIA-DEP59 which therefore warrants a further examination. In the sampling site, our specimens were collected from soil mixed

with leaves litter. Larsson et al. (2009) emphasized that many Inocybe species in the section Rimosae (currently genus Inosperma) are known to occur on more nutrient rich soil. In addition, I. sphaerobulbo-sum FIPIA-DEP59 grows near Euphorbiaceae and Arecaceae which dominate the surrounding area and in contrast to Deng et al. (2021) reported the specimens were collected near fagaceous trees. The Inocybaceae is known as family of ectomycorrhizal mushroom-forming fungi (Matheny et al., 2019), however the host plant of I. sphaerobulbosum FIPIA-DEP59 remains unclear.

In the field, I. sphaerobulbosum FIPIA-DEP59 can be recognized by its gregarious habit, medium-sized basidioma, having scaly-fibrillose pileus, and a fibrillose stipe. Inosperma sphaerobulbosum FIPIA-DEP59 has approximately similar of pileus dimension to I. carnosibulbosum (Pradeep et al., 2016) but larger in size when compared to I. akirnum, and I. saragum Latha and Manimohan (2017). Some ba-

sidiomata of I. sphaerobulbosum FIPIA-DEP59 displays pileus with layer of universal veil. Deng et al. (2021) emphasized that this feature is non persistent character and is influenced easily by the weather. The lamellar edge of I. sphaerobulbosum FIPIA-DEP59 was fimbricate and slightly serrate which in contrast to the related morphological similar species in having only fimbricate edge. Microscopically, the outline of basidiospores of I. sphaerobulbosum FIPIA-DEP59 differs it to the closely morphological species of I. carnosibulbosum, I. akirnum, and I. saragum. The spore dimension of our specimen was slightly lower than the three closely related species. However, in line with those species, I. sphaerobulbosum FIPIA-DEP59 has abundant cheilocystidia in clusters. In addition, the yellow intracellular contents of cheilocystidia distinguished our specimen from the genus Pseudosperma.

A comparison of the ITS sequence of specimen FIPIA-DEP59 derived from Central Kalimantan with the available taxa information from GenBank

indicated that BLAST queries led to I. sphaerobulbosum from type material as the top hits. Accordingly with BLAST result, the phylogenetic tree committed that our Inosperma was identified as I. sphaerobulbosum. The phylogenetic placement of our specimen in the same clade as I. sphaerobulbosum from Hainan, China with 93 % BS value. In the phylogenetic tree, the position of clade of I. sphaerobulbosum FIPIA-DEP59 was mixed between Old World tropical clade 1 and Old World tropical clade 2 (Pradeep et al., 2016; Latha, Manimohan, 2017; Matheny et al., 2019; Aïgnon et al., 2021) due to only single gene used in this study. Previous study confirmed that Old World tropical clade 2 was composed of Inosperma species from India, Thailand, Papua New Guinea, and China (Matheny et al., 2019; Deng et al., 2021). Inosperma sphaerobulbosum is reported for the first time for Indonesia and enriches the knowledge of the distribution of Old World tropical clade 2. Our study provides the available morphological and molecular information of I. sphaerobulbosum in Indonesia.

Fig. 3. Microscopic characters of Inosperma subsphaerosporum FIPIA-DEP59: A - globose to subglobose basidiospores; B - cheilocystidia in clusters; C - basidia; D - basidia with basidiospores. Bars = 20 ^m.

Fig. 5. Microscopic characters of Inosperma subsphaerosporum FIPIA-DEP59: A - pileipellis; B - pileal trama; C, D -oleiferous hyphae; E - clamp connection; F - stipe trama. Bars A = 200 ^m, B, C, F = 50 ^m, D, E = 20 ^m.

Fig. 4. Scanning electron microscope image of Inosperma subsphaerosporum FIPIA-DEP59: A, B -basidiospores with an indentation; C - cystidium; D - basidium with four sterigmata.

88 ^

•Inosperma apiosmotum strain PBM3020 JQ801385 •Inosperma aff. calamistratum strain REH8420 JQ801390 1 Inosperma mucidiolens voucher WTU:DG1824 HQ201339 Inosperma calamistratum strain PBM1105 JQ801386

-Inosperma misakaense strain PC96234 JQ801409 Inosperma cyanotrichium strain 137 JQ801396 Inosperma calamistratoides strain PBM3384 JQ801393 \ Inosperma rubricosum strain PBM3784 KP308817 Inosperma viridipes voucher HO-562693 KP641646 Inosperma latericium voucher PDD-92382 GU233367 Inosperma cervicolor voucher TURA-4761 JQ801395 Inosperma monastichum voucher STU-SMNS-STU-F-0901533 MW647631 ■ Inosperma aff. calamistratum DED8134 GQ892983 Inosperma aff. latericium strain TR109-02 JQ801405 Inosperma vinaceum voucher AMB-18747 MW561108

loo i Inosperma bulbomarginatum voucher HLA0417 MT534300

'Inosperma bulbomarginatum voucher MR00357 MN096190 Inosperma akirnum voucher CAL1358 KY440085

— Inosperma saragum voucher CAL1360 KY440103

— Inosperma carnosibulbosum TBGT12047 KT329448 791 Inosperma virosum TBGT753 KT329452

■ Inosperma gregarium voucher CAL1309 KX852305 'Inosperma hainanense voucher Zeng4937 MZ374070 Inosperma hainanense voucher Zeng4936 MZ374069 Inosperma muscarium voucher Zeng4720 MZ373978 Inosperma muscarium voucher Zeng4736 MZ373979

Inosperma flavobrunneum voucher HLA0372 MT534290

10i

T-| Inosperma subsphaerosporum voucher FHMU3154 MW403826 Inosperma subsphaerosporum voucher FHMU3153 MW403825 Inosperma subsphaerosporum voucher FIPIA-DEP59 OP880257 Inosperma subsphaerosporum voucher FHMU3155 MW403827 loo I Inosperma africanum voucher MR00387 MN096189 Inosperma africanum voucher HLA0383 MT534298 Inosperma changbaiense voucher HMJAU25861 MH047251 Inosperma cf. lanatodiscum TURA1812 JQ408763 Inosperma aff. maculatum isolate AFTOL-ID-476 DQ241778 Inosperma neobrunnescens strain SAT0427406 JQ801411 Inosperma ismeneanum voucher STU-SMNS-STU-F-0901561 MW647625 Inosperma dodonae voucher STU-SMNS-STU-F-0901253 MW647615 i—Inosperma bicoloratum voucher ZT12187 GQ892984 '— Inosperma aff. fastigiellum strain PBM3325 JQ801399 Inosperma rimosoides isolate AFTOL-ID-520 DQ404391 Auritella hispida strain TH1009 KT378203

Fig. 6. Inosperma subsphaerosporum FIPIA-DEP59 phylogenetic tree based on ITS1-2 region using Randomized Axe-lerated Maximum Likelihood method and 1000 Bootstrap Analysis. Our specimen is bold on the phylogenetic tree.

In the research site, I. sphaerobulbosum is considered as delicacy mushroom and traded at local market as well as by using the social media. The mushroom is prized Rp. 18000/kg or US$1.15/kg.

The indigenous people usually forage this species around September to December, and sometimes March to April when the rain and relative humidity are high. Considering the edibility of this species, we

suggest that the nutritional analysis and cultivation efforts of I. sphaerobulbosum needs a warrant in Indonesia, as many wild mushrooms are recognized as source of secondary metabolite and medicinal properties (Bukharova et al., 2020). However, many species of Inosperma were reported to cause neurotoxic poisoning in humans after consumption worldwide (Bijeesh et al., 2020; Parnmen et al., 2021; Deng et al., 2022; Li et al., 2022). For example, I. erubescens caused mass poisoning in Germany (Lurie et al., 2009), 10 cases of poisoning caused by Inosperma species in Thailand from 2010 to 2018 (Parnmen et al., 2021), and recently I. zonativeliferum caused poisoning in 10 people in China (Deng et al., 2022). Li et al. (2022) reported that I. sphaerobulbosum contains none of the tested neurotoxins. Although no report indicates the poisoning case by consumption of this mushroom from type locality as well as in our sampling site, we expect to determine the four main mushroom neurotoxins (muscarine, psilocybin, ibo-tenic acid, and muscimol) contents of I. sphaerobul-bosum FIPIA-DEP59 and closely related species in the near future.

Conclusion

The current study reports the first distribution of Inosperma subsphaerosporum in Indonesia and new information aside from type locality (China).

Morphologically, I. sphaerobulbosum FIPIA-DEP59 can be determined by the conical applanate cap with umbo, incurved to decurved margin, dry pileus with fibrillose to rimulose surface, lamellae adnexed to sinuate, cylindrical stipe, subglobose to globose smooth basidiospores, and abundant cheilocystidia in clusters. The BLAST result and the phylogenetic tree confirmed our specimens as I. subsphaerospo-rum. Future study should be done to analyze both nutritional and mushroom neurotoxins content of this Indonesian Inosperma.

Acknowledgements

This study was supported by the grant of "Dana Masyarakat IPB 2023 - Penelitian Dosen Muda" with reference number 11401/IT3/PT.01.03/P/B/2023 to Dr. Ivan Permana Putra. We would like to thank Mr. Andi for the field assistance and Mr. Rudy for the fruitful discussion. We thank the Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Indonesia, The Research Center for Applied Microbiology, National Research and Innovation Agency (BRIN), Bogor, West Java, Indonesia, The Department of Parasitology, Faculty of Medicine, Palangka Raya University, Central Kalimantan, Indonesia and the KPJI. We thank the Herbarium Bandungense, Indonesia for the herbarium support.

REFERENCES / ЛИТЕРАТУРА

Aïgnon H. L., Jabeen S., Naseer A., Yorou N. S., Ryberg M. 2021. Three new species of Inosperma (Agaricales, Inocybaceae) from Tropical Africa. MycoKeys 77: 97-116. DOI: 10.3897/mycokeys.77.60084

Bijeesh C., Pradeep C. K., Vrinda K. B. 2020. Inocybe poisoning from Kerala-a case study. J. Mycopathol. Res. 57(4): 255-258.

Bukharova N. V., Prozorova L. A., Ngô X. Q. 2020. The first data on wood-inhabiting fungi (Basidiomycota) of the Con Dao Archipelago (South Vietnam). Turczaninowia 23, 3: 83-91. DOI: 10.14258/turczaninowia.23.3.8

Deng L., Yu W., ZengN., Liu L., Liu L., Fan Y. 2021. Inosperma subsphaerosporum (Inocybaceae), a new species from Hainan, tropical China. Phytotaxa 502: 169-178. DOI: 10.11646/phytotaxa.502.2.5

Deng L. S., Yu W. J., Zeng N. K., Zhang Y. Z., Wu X. P., Li H. J., Xu F., Fan Y. 2022. A new muscarine-containing Inosperma (Inocybaceae, Agaricales) species discovered from one poisoning incident occurring in tropical China. Front. Microb. 13: 923435. DOI: https://doi.org/10.3389/fmicb.2022.923435

Goldstein J. I., Newbury D. E., Echlin P., Joy D. C., Romig Jr. A. D., Lyman C. E., Fiori C., Lifshin E. 1992. Scanning electron microscopy and X-ray microanalysis, 2nd ed. New York: Plenum Press. 840 pp.

Index Fungorum [2023]. Inosperma. http://www.indexfungorum.org/Names/Names.asp (Accessed 01 June 2023). Kropp B. R., Matheny P. B., Hutchison L. J. 2013. Inocybe section Rimosae in Utah: phylogenetic affinities and new species. Mycologia 105: 728-747. DOI: 10.3852/12-185

Kühner R. 1980. Les Hyménomycètesagaricoïdes (Agaricales, Tricholomatales, Plutéales, Russulales). Etude générale et classification. Bulletin de la Société Linnéenne de Lyon 50(4): 1-1927.

Larsson E., Ryberg M., Moreau P. A., Mathiesen A. D., Jacobsson S. 2009. Taxonomy and evolutionary relationships within species of section Rimosae (Inocybe) based on ITS, LSU and mtSSU sequence data. Persoonia 23: 86-98. DOI: 10.3767/003158509x475913

Latha K. P. D., Manimohan P. 2017. Inocybes of Kerala. Calicut: SporePrint Books. 181 pp.

Li S. N., Xu F., Long P., Liu F., Zhang P., Fan Y., Chen Z. 2022. Five new species of Inosperma from China: Morphological characteristics, phylogenetic analyses, and toxin detection. Front. Microbiol. 13: 1021583. DOI: 10.3389/ fmicb.2022.1021583

Lurie Y., Wasser S. P., Taha M., Shehade H., Nijim J., Hoffmann Y., Basis F., Vardi M., Lavon O., Suaed S., Bi-sharat B., Bentur Y. 2009. Mushroom poisoning from species of genus Inocybe (fiber head mushroom): A case series with exact species identification. Clin. Toxicol. 47(6): 562-565. DOI: 10.1080/15563650903008448

Matheny P. B., Hobbs A. M., Esteve-Raventos F. 2019. Genera of Inocybaceae: New skin for the old ceremony. Mycologia 112: 83-120. DOI: 10.1080/00275514.2019.1668906

Parnmen S., Nooron N., Leudang S., Sikaphan S., Polputpisatkul D., Pringsulaka O., Binchai S., Rangsiruji A. 2021. Foodborne illness caused by muscarine-containing mushrooms and identification of mushroom remnants using phylogenetics and LC-MS/MS. Food Control 128: 108182. DOI: 10.1016/j.foodcont.2021.108182

Pradeep C. K., Vrinda K. B., Varghese S. P., Korotkin H. B., Matheny P. B. 2016. New and noteworthy species of Inocybe (Agaricales) from tropical India. Mycol. Progr. 15: 1-25. DOI: 10.1007/s11557-016-1174-z

Putra I. P., Hafazallah K. 2020. Catatan Komunitas Pemburu Jamur Indonesia: Kolaborasi Lintas Profesi dan Ge-nerasi Mengenai Etnomikologi Jamur-Jamur Indonesia. Sukabumi: Haura Publishing. 62 pp. [In Bahasa] Singer R. 1975. The Agaricales in modern taxonomy 3rd ed. Vaduz: J. Cramer. 912 pp.

Stamatakis A. 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bio-informatics 30: 1312-1313. DOI: 10.1093/bioinformatics/btu033

White T. J., Bruns T., Lee S. J., Taylor J. W. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: M. Innis, D. Gelfand, J. Sninsky, T. White (eds.). PCR Protocols. New York: Academic Press. Pp. 315-322.