Newly Recorded Panus lecomtei (Fr.) Corner (Basidiomycota: Panaceae) for Indonesia

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Makara Journal of Science Makara Journal of Science

Volume 27

Issue 3 September Article 1

9-25-2023

Newly Recorded Panus lecomtei (Fr.) Corner (Basidiomycota:

Panaceae) for Indonesia Panaceae) for Indonesia

Ivan Permana Putra

Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor 16680, Indonesia, [email protected]

Rudy Hermawan

Alumni of Master Program of Microbiology, Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor 16680, Indonesia

Ali Bin Abithalib Salman

Alumni of Master Program of Microbiology, Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor 16680, Indonesia

Follow this and additional works at: https://scholarhub.ui.ac.id/science Part of the Biodiversity Commons

Recommended Citation Recommended Citation

Putra, Ivan Permana; Hermawan, Rudy; and Salman, Ali Bin Abithalib (2023) "Newly Recorded Panus lecomtei (Fr.) Corner (Basidiomycota: Panaceae) for Indonesia," Makara Journal of Science: Vol. 27: Iss. 3, Article 1.

DOI: 10.7454/mss.v27i3.1507

Available at: https://scholarhub.ui.ac.id/science/vol27/iss3/1

This Article is brought to you for free and open access by the Universitas Indonesia at UI Scholars Hub. It has been accepted for inclusion in Makara Journal of Science by an authorized editor of UI Scholars Hub.

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Newly Recorded Panus lecomtei (Fr.) Corner (Basidiomycota: Panaceae) for Indonesia

Ivan Permana Putra

^1*

, Rudy Hermawan

²

, and Ali Bin Abithalib Salman

²

1. Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor 16680, Indonesia 2. Alumni of Master Program of Microbiology, Department of Biology, Faculty of Mathematics and Natural Sciences,

IPB University, Bogor 16680, Indonesia

*E-mail: [email protected]

Received Februari 6, 2023 | Accepted June 13, 2023

Abstract

Panus lecomtei (Polyporales) is reported for the first time from Indonesia at the IPB University Campus Forest (IPBUCF).

This species has similar macroscopic features to P. strigellus and P. neostrigosus. This study aimed to confirm the taxonomic position of P. lecomtei found at IPBUCF based on morphological and molecular evidence. The specimens were described by their macroscopic and microscopic characteristics. A phylogenetic approach was used with the internal transcribed spacer 1/2 sequence data. The combination of morphological and molecular analyses confirmed the taxonomic position of our specimens as P. lecomtei. The recorded species was distinguished by gregarious basidiomata, an infundibuliform resembling a funnel, violet during the immature stage to orangish brown in the mature pileus and ellipsoid basidiospore stages. Our specimens were distinguished from P. strigellus by having smaller basidiospores. It was difficult to separate P. lecomtei BO24428 from P. neostrigosus based on morphological characters. However, the BLAST results showed that specimen BO24428 had 99.56% similarity with P. lecomtei. In addition, our phylogenetic tree nested P.

lecomtei BO24428 in the P. lecomtei clade. Morphological illustrations and a phylogenetic tree of P. lecomtei are provided.

We suggest combining the morphological and molecular evidence in future studies of P. lecomtei from Indonesia.

Keywords: macrofungi, phylogeny, polyporales, taxonomy, West Java

Introduction

The genus Panus is distributed worldwide from temperate to tropical regions [1–3]. This lentinoid fungi is recognized by having free gills and has been demonstrated to be monophyletic with Lentinus and Polyporus [4], regardless of the lack of a gill structure in the latter genus.

Previous reports have indicated that Panus is classified together with Lentinus and Polyporus in the family Polyporaceae [2, 4, 5]. However, the Index Fungorum records Panus in the family Panaceae [6]. To date, 236 Panus taxa are listed in the Index Fungorum [6].

Until 2017, the Indonesian Institute of Sciences reported 2,273 species of (macro-and microscopic) fungi in Indonesia, or barely about 0.0015% of the total expected number of fungal species worldwide [7]. Macrofungal diversity is not well studied in Indonesia, including that in the genus Panus. The only proper documentation for Panus was provided by Susan and Retnowati [8], who reported P. neostrigosus from Enggano Island. The observation was based only on the macroscopic

characters of the specimen. The morphological characters are the basic data needed to identify macrofungi despite their variability [9, 10]. However, some Panus species are morphologically similar, including P. neostrigosus, P. strigellus, and P. lecomtei [1].

P. lecomtei (Fr.) was proposed by Corner in 1981 based on L. lecomtei constructed by Fries. This species is recognized as an important edible mushroom because of its useful nutritional and medicinal compounds [11].

Although this species is distributed in subtropical and tropical areas [1], no study has described the distribution of P. lecomtei in Indonesia until now. Hence, we explored this species in the Campus Forest of IPB University (Indonesia), as a previous report showed a high diversity of macrofungi in this area [12]. During the survey, some purple (immature stage) and brown (mature stage) lentinoid fruiting bodies that grew on decaying trunks were collected. The specimen was initially identified as P. lecomtei but this study aimed to ensure the taxonomic position of P. lecomtei from Indonesia based on morphological and molecular evidence.

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Materials and Methods

Specimen collection. The fruiting bodies were collected in February 2022 at IPB University Campus Forest, West Java, Indonesia. All basidiomata stages that grew in the same spot were collected. The macrofungi were documented in situ and in the laboratory. Further investigations were conducted in the Laboratory of Mycology, Department of Biology, IPB University, Indonesia. The fruiting bodies were deposited at the Herbarium Bogoriense Indonesia under collection number BO24428.

Morphological identification. The morphological characters were recorded, including size, color, shape, ornamentation, and spores. The features were observed using stereo and binocular microscopes (400 and 1,000×). The samples were mounted in distilled water.

The methods for morphological characterization and identification followed several publications [1, 2, 13, 14].

DNA extraction, polymerase chain reaction (PCR), and sequencing. Genomic DNA was extracted using a protocol published previously [15]. DNA quality and quantity were verified using a Nanodrop spectro- photometer. The internal transcribed spacer (ITS) region was used for amplification. The primers were ITS 5 (5′- GGA AGT AAA AGT CGT AAC AAG G-3′) and ITS 4 (5′-TCC TCC GCT TAT TGA TAT GC-3′) [16]. PCR amplification was performed in a total reaction volume

of 40 µL containing 12 µL ddH2O, 2 µL of 10 pmol of each primer, 20 µL of the PCR mix from 2X Kappa Fast 2G, and 4 µL of 100 ng template DNA. The PCR conditions for the ITS primers followed a previous protocol [17]. PCR amplification was performed in a 40 µL total reaction volume containing 20 µL of PCR mix from 2× Kappa Fast 2G, 2 µL (10 pmol) of each primer, 4 µL (100 ng) of template DNA, and 12 µL of ddH2O.

The PCR conditions were initial denaturation at 94 °C for 2 min, followed by 30 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C.

The amplicons were checked by 1.5% agarose gel electrophoresis and visualized with the Gel Doc^TM XR system. PCR products were sent to the First Base Malaysia for sequencing. The nucleotide sequence was deposited in GenBank to acquire the accession number.

Phylogenetic analyses. A phylogenetic tree was constructed from 25 datasets (Table 1), including the sequence from the current study (bold), 23 sequences of Panus species from [2], and L. Squarrosulus as an outgroup from [17]. The alignment and phylogenetic analyses used previous protocols. Clustal X software [18]

was used for the sequence alignment and was transformed into a PHYLIP format file. The phylogenetic tree was constructed using the Randomized Accelerated Maximum Likelihood Black Box, generated on CIPRES [19]. The phylogenetic tree was assessed with 1,000 bootstrap replicates. Bootstrap values ≥ 50 were displayed on the phylogenetic tree branch.

Table 1. Species, Collection Codes, and GenBank Accession Numbers used in this Study

Species Collection code Gene acc. no

ITS

Panus conchatus X1234 JN710579

Panus conchatus KUMCC18-0047 Mk192053

Panus conchatus JMH44 KM267730

Panus conchatus FLAS-F-60901 MH016880

Panus conchatus CLZhao 1535 MG231759

Panus lecomtei BO24428 OL763317

Panus lecomtei HHB-9614 KP135329

Panus lecomtei HHB-11042-Sp KP135328

Panus lecomtei TMIC35103 JQ955726

Panus neostrigosus LSPQ-NSM-107 KU761235

Panus rudis ZJ1005DKJ04 KU863051

Panus similis LE287548 KM411466

Panus similis KWGM 39 KY630517

Panus sp. MEL 2382698 KP012877

Panus sp. MEL 2382967 KP012827

Panus sp. M85 KP096364

Panus strigellus TENN55993 JQ955728

Panus strigellus INPA239979 JQ955724

Panus strigellus INPA222827 JQ955722

Panus strigellus TENN56192 JQ955727

Lentinus squarrosulus BO 24427 MT815466

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Results and Discussion

Taxonomy. Panus lecomtei (Fr.) Corner, Beih. Nova Hedwigia 69: 90 (1981) (Figures 1–3).

Basionym:

Lentinus lecomtei Fr. 1825

GenBank Submission: ITS OL763317.

Cap 2–9 cm in diameter, gregarious, dry, umbilicate to infundibuliform resembling a funnel, inrolled margin, with a range of colors from violet during the immature stage to reddish or orangish brown at maturity, leathery appearance, covered with dense tiny hairs over the

surface. Lamellae decurrent and intensively running down the stem at maturity, medium to crowd, straight, intercalated with some lamellulae near the margin, whitish to cream. Stipe 1–3.8 cm in length, 0.4–1.2 cm in width, tough, off-central to near lateral of the pileus, cylindrical to slightly inflated at the base, pale violet during the immature stage to light orangish brown at maturity, densely hairy with white hair color in the immature phase to brown in the mature fruiting body.

Basidia 3.1–3.3 µm in width, clavate, tetrasporic. Spores 4.2–5.3 × 1.5–1.9 µm, ellipsoid, smooth, and hyaline.

Gloeocystidia absent. No metuloids were observed.

Hyphal system monomitic, pileipellis a cutis, stipe tissue compact and arranged in parallel.

Figure 1. The Fruiting Bodies of Panus lecomtei BO24428. A. Several Stages of the Fruiting Body on Substrate. B. Development of the Fruiting Body. Bars: A = 10 cm, B = 5 cm

Figure 2. Macromorphological Characteristics of Panus lecomtei BO24428. A. Cap Surface Ornamentation of Immature Fruiting Body. B. Lamellae of Immature Fruiting Body. C. Cap Surface Ornamentation of Mature Fruiting Body.

D. Lamellae of Mature Fruiting Body. E. Stipe Ornamentation. Bars: A–E = 5 cm

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168 Putra, et al.

Figure 3. Micromorphological Characteristics of Panus lecomtei BO24428. A. Pileipellis. B. Hyphae of Stipe. C. Basidium with 4 Sterigmata (Arrow). G. Basidiospores (Arrow). Bars: A = 10 µm, B = 100 µm, C, D = 5 µm

Habitat: Decaying trunk in a small forest.

Specimen examined: Indonesia, West Java, Bogor, IPBUCF, 6°33'18.6"S 106°43'17.4"E, 197 m a.s.l., February 2022, collected by Salman ABA, BO24428.

Distribution: Asia, Africa, Europe, North America, Central America, South America, Australia, and Oceania.

Phylogenetic Analyses. The sequence was submitted to GenBank (https://www.ncbi.nlm.nih.gov/) with Accession number ITS OL763317. The BLAST result showed that the ITS sequence of our specimens had 99.56% similarity with P. lecomtei as the top hit. The phylogenetic tree (Figure 4) provided high support that Panus BO24428 was P. lecomtei. The phylogenetic tree divided some specimens into species clades. There were 6 clades, i.e., Panus similis, P. rudis, P. neostrigosus, P. lecomtei, P.

strigellus, and P. conchatus. Our specimen was nested in the P. lecomtei clade.

P. lecomtei (family Panaceae) has never been reported in Indonesia. The majority of Panus reports in Indonesia were based on macroscopic features and frequently lacked important morphological information. To date, the report of Panus in Indonesia only includes P. neostrigosus.

In addition, only 3 studies [8, 20, 21] have provided macroscopic documentation and a morphological description of the genus Panus (P. neostrigosus). Several Panus species may be confused macroscopically with P.

lecomtei [1]. Our specimens were morphologically identical to Indonesian P. neostrigosus, previously reported based on morphological characters [8]. P. neostrigosus has dense hair on the surface of the brown pileus and it is difficult to distinguish from P. lecomtei. Kuo [22]

reported that the fruiting bodies of both species are purple at the immature stage, and fade to reddish brown, pinkish brown, orangish brown, or tan at maturity. In addition, P.

lecomtei and P. strigellus have the same color and shape of the pileus, and ovoid to ellipsoid spores [1]. Our investigation of the morphological characters confirms that our specimen was matched to P. lecomtei based on the descriptions in [1] and [23], as it had slightly smaller basidiospores than P. strigellus.

P. lecomtei and P. strigellus are morphologically identical; therefore, they are easily confused [1]. In addition, P. lecomtei BO24428 differs morphologically based on the color of the basidiomata, stipe size, spore shape, and spore size from P. lecomtei described from

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Piauí State, Brazil [3]. This result suggests the morphological plasticity of the genus Panus, as pointed out by [1]. Hussein [4] suggested that Panus and Lentinus form a monophyletic clade with Polyporus. This was consistent with Kuo [24], who explained that they evolved together. Due to the plasticity of morphological characters between Panus species and other Indonesian mushroom identification cases [25], it is crucial to support the description with molecular data to obtain a more accurate taxonomic identity. Therefore, in this study, the morphological data coupled with DNA analyses confirmed the identity of P. lecomtei. To the best of our knowledge, this is the first record of P.

lecomtei from Indonesia. The results of this study will be helpful for the Panus database in Indonesia.

In this study, we decided not to use the P. lecomtei ITS sequence from the BLAST result for the phylogenetic tree, but we used the sequences from Luangharn [2], as the top BLAST hits were unpublished sequences.

Hitherto, Index Fungorum accepted P. neostrigosus as the current name of P. lecomtei [26] based on Drechsler- Santos [27]. However, we treated our specimen as P.

lecomtei based on evidence from the phylogenetic tree (Figure 4). The clade was clearly separate from the

morphologically similar P. strigellus, P. neostrigosus, and other clades. In line with our result, Vargas-Isla [1]

considered P. lecomtei as the priority name and P.

neostrigosus as a synonym of the former. In addition, Luangharn [2] reported a phylogenetic tree inferred from ITS and LSU sequences in which P. lecomtei and P.

neostrigosus were in the same clade. However, they decided to mark this as the lecomtei clade. In our phylogenetic tree, P. neostrigosus was in a different clade from P. lecomtei. In addition, P. Lecomtei was a sister clade of P. rudis and P. neostrigosus (Figure 4).

We suggest that P. neostrigosus, which was previously reported from Indonesia, might be P. lecomtei. Due to the high plasticity in the morphological features of Panus species, we strongly recommend combining morphological and molecular analyses to confirm the taxonomic position of Panus species in Indonesia. In addition, identification using phenotypic and gene characters should be considered for a reliable taxonomic framework of macrofungi in Indonesia. Our P. lecomtei ITS data (OL763317) are now available in Genbank and can be used for further Panus research to provide accurate and precise identifications of Indonesian macrofungi.

Figure 4. Phylogenetic Tree Inferred by the ITS1/2 using RAxMLBlack Box. Newly Recorded Species is Indicated in Bold.

Bootstraps (BS) ≥ 50 are Displayed on the Branches

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Conclusion

The current study reports a new record of Panus lecomtei from Indonesia based on morphological and molecular data. The combination of morphological and molecular evidence confirmed the identity of P. lecomtei BO24428.

The smaller basidiospores of our specimens were distinguished from P. strigellus. The morphological characters made it difficult to separate our specimens from the morphologically similar species P. neostrigosus.

The BLAST results confirmed that P. lecomtei BO24428 had 100% similarity with P. lecomtei. In addition, our phylogenetic tree nested P. lecomtei BO24428 in the P.

lecomtei clade. We suggest that morphological data should be combined with molecular analysis in future studies of P. lecomtei in Indonesia.

Acknowledgment

We are grateful to the Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, for the research facilities. In addition, we thank Direktorat Pengelolaan Koleksi Ilmiah (BRIN), Indonesia for the herbarium support.

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