Hidden in the daylight: A polyphasic approach reveals two bioluminescent Mycenaceae species from Yunnan Province, China (Basidiomycota, Agaricales)

Biodiversity Data Journal 13: e168858 doi: 10.3897/BDJ.13.e168858 Taxonomy & Inventories Hidden in the daylight: A polyphasic approach reveals two bioluminescent Mycenaceae species from Yunnan Province, China (Basidiomycota, Agaricales) Wenhua Lu , Alanoud T. Alfagham , Saowaluck Tibpromma , Nakarin Suwannarach , Jaturong Kumla , Dong-Qin Dai , Abdallah M. Elgorban , Ekachai Chukeatirote , Samantha C. Karunarathna ‡ Center for Yunnan Plateau Biological Resources Protection and Utilization & Yunnan International Joint Laboratory of Fungal Sustainable Utilization in South and Southeast Asia, College of Biology and Food Engineering, Qujing Normal University, Qujing 655099, China § Excellence Center of Microbial Diversity and Sustainable Utilization, Chiang Mai University, Chiang Mai 50200, Thailand | Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand ¶ Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia # Office of Research Administration, Chiang Mai University, Chiang Mai 50200, Thailand ¤ Center of Excellence in Biotechnology Research (CEBR), King Saud University, Riyadh 11451, Saudi Arabia « School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand Corresponding author: Ekachai Chukeatirote ([email protected]), Samantha C. Karunarathna ([email protected]) Academic editor: Olivier Raspé Received: 15 Aug 2025 | Accepted: 11 Dec 2025 | Published: 18 Dec 2025 Citation: Lu W, Alfagham AT, Tibpromma S, Suwannarach N, Kumla J, Dai D-Q, Elgorban AM, Chukeatirote E, Karunarathna SC (2025) Hidden in the daylight: A polyphasic approach reveals two bioluminescent Mycenaceae species from Yunnan Province, China (Basidiomycota, Agaricales). Biodiversity Data Journal 13: e168858. https://doi.org/10.3897/BDJ.13.e168858 Abstract Background Yunnan Province, recognised as a biodiversity hotspot in China, is a rich source of fungal diversity that we are just beginning to explore. During our investigation of bioluminescent mushrooms in this region, we collected four specimens from the family Mycenaceae. We ‡,§,| ¶ ‡ §,|,# §,|,# ‡ ¤ « ‡ © Lu W et al. 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. conducted morphological characterisation, multilocus phylogenetic analyses (ITS, LSU, SSU, tef1-α and rpb2) and photographed specimens under completely dark conditions. New information In this paper, we report on two bioluminescent mushrooms of Mycenaceae collected in Yunnan Province, China. Morphological and phylogenetic analyses revealed Mycena semivestipes, which was first observed with bioluminescence and Roridomyces pruinosoviscidus, which has not been previously recorded in China. Additionally, we provide descriptions, illustrations, phylogenetic analysis results and photographs that highlight the bioluminescent features of these taxa. This study enhances our understanding of bioluminescent mushroom diversity in the Yunnan Province, raising the total number of bioluminescent mushrooms reported from China to 36. Keywords bioluminescent fungi, diversity, ecology, Mycena semivestipes, Roridomyces pruinosoviscidus, saprobic fungi Introduction Yunnan Province, situated in southwest China, is one of the regions with the richest biodiversity in the country. Its unique geographical environment and climatic conditions support a rich and diverse array of biological resources (Zhang et al. 2021). However, despite the increasing research on plant and animal diversity, the understanding of fungal diversity remains understudied (Feng and Yang 2018, Lu et al. 2024a). Bioluminescent fungi, a remarkable group of organisms within the fungal kingdom, have garnered significant attention for their potential applications in ecology, medicine and genetic engineering (Syed and Anderson 2021, Lu et al. 2024b). Bioluminescence is not uncommon, but its occurrence in fungi is relatively rare. Fungal bioluminescence is produced by a special biochemical reaction, which usually involves luciferin, luciferase and oxidation (Kotlobay et al. 2018, Lu et al. 2024b). Kotlobay et al. (2018) reported that the fungal bioluminescence biosynthesis pathway involves fungal luciferase and three other key enzymes in a cycle of caffeic acid. The new view emerged by integrating gene editing technology with plant cultivation, enabling the transfer of mushroom genes into living organisms (Mitiouchkina et al. 2020, Zheng et al. 2023, Ge et al. 2024). However, the role of mushroom bioluminescence in ecology remains a mystery. First, some researchers believe that fungi emit light to attract phototactic insects, thereby assisting in spore dispersal, especially in deep forest areas where wind is blocked (Lu et al. 2024b). Creating artificial mushrooms with light has been shown to attract more insects than non-luminous mushroom models (Oliveira et al. 2015). Weinstein et al. (2016) scrutinised the abundance of insects recorded in the field during basidiomata 2Lu W et al production and they evaluated the allure of bioluminescent fungi to flying insects, finding no discernible difference in Omphalotus nidiformis (Berk.) O.K. Mill. Second, the bioluminescence might arise as an incidental by-product of metabolism rather than providing any distinct evolutionary advantage (Nimalrathna et al. 2022, Lu et al. 2024b). The bioluminescent parts vary significantly amongst different species; Armillaria (Fr.) Staude species exhibit bioluminescence during the mycelium stage, while in other species, bioluminescence occurs on the pileus, gills, stipe or spores (Lu et al. 2024b). During the development of Armillaria from mycelium to mushroom, it has been observed that the synthesis of luciferin precursors and hispidin-3-hydroxylase was inhibited (Mihail et al. 2018). Additionally, some studies hypothesise whether bioluminescent mushrooms generate oxidative stress responses to counteract the oxygen released during the respiratory phase, as the bioluminescent mechanism also entails oxygen consumption (Briones-Martin-Del-Campo et al. 2014, Warris and Ballou 2019, Yaakoub et al. 2022, Emri et al. 2024). Moreover, all bioluminescent mushrooms are found in tropical, subtropical and temperate regions (Lu et al. 2024b). Mushrooms dissipate excess absorbed energy to maintain internal energy balance. Bioluminescence — often referred to as 'cold light' (Cordero et al. 2023) — may serve this function, suggesting it is an evolutionary adaptation for energy regulation. This phenomenon ensures that mushrooms do not overheat due to absorbed energy, which could potentially harm their cellular structures or disrupt metabolic processes (Iqbal et al. 2009, Tisch and Schmoll 2010). Instead, they convert this excess energy into visible light, a process crucial for survival in their respective habitats. However, further investigation is necessary to elucidate the significance of the mushroom bioluminescence. There are more than 132 fungal species that exhibit bioluminescence properties, all of which belong to the Basidiomycota, except for Xylaria hypoxylon (L.) Grev, which belongs to the phylum Ascomycota and, amongst them, the family Mycenaceae Overeem contains most bioluminescent mushrooms worldwide (Lu et al. 2024b, Perry et al. 2025). Mycena (Pers.) Roussel is the largest genus in Mycenaceae, comprising more than 500 species distributed worldwide (Nagamune et al. 2024, Soares et al. 2024). It plays a key role in the circulation of forest ecosystems, as saprobic fungi dominate the degradation of fibres synergistically with endophytic fungi and some species contain biologically active and antimicrobial compounds (Bäuerle et al. 1982, Frankland 1998, Fukasawa et al. 2009). The genus Mycena is characterised by small, multicoloured basidiomata; pileus conical, parabolic and bell-shaped, hygrophanous, membranous with or without striations on the surface and smooth; lamellae adnate, decurrent or arcuate; stipe cylindrical, hollow and fragile; cystidia with protuberances, various shapes, clavate, utriform, pyriform, fusiform, hyaline, amyloid and thin-walled, containing oil; and spores globose, subglobose, ellipsoid, cylindrical and occasionally ovoid (Seok et al. 2015, Qiang and Bai 2023, Nagamune et al. 2024, Wei et al. 2024). This genus comprises more than 80 described or reported bioluminescent mushroom species, the largest number of bioluminescent species known worldwide (Chew et al. 2014, Heinzelmann et al. 2024, Lu et al. 2024b, Soares et al. 2024, Perry et al. 2025). Hidden in the daylight: A polyphasic approach reveals two bioluminescent ... 3 Roridomyces Rexer was established by Rexer (Rexer 1994) with the type species R. roridus (Fr.) Rexer, characterised by pileus trama, composed of interwoven, cylindrical hyphae and the pileipellis, a hymeniderm composed of clavate to subglobose terminal elements. There are 15 species worldwide; amongst them, six are bioluminescent mushrooms. Seven Roridomyces species have been reported from China, viz. R. appendiculatus Rexer, R. glutinosus (Corner) T. Bau & L.N. Liu, R. mauritianus (Robich & Hauskn.) Hauskn. & Krisai, R. praeclarus (E. Horak) Rexer, R. lamprosporus (Corner) Rexer, R. roridus and R. viridiluminus L.A.P. Dauner, Karunarathna & P.E. Mortimer (Dauner et al. 2021), of which the last three species are bioluminescent (Dauner et al. 2021, Lu et al. 2024b). As mentioned above, the peculiar biological phenomenon of bioluminescence is of great significance in biological research and has attracted widespread interest. For this reason, understanding the diversity of bioluminescent fungi and their ecological characteristics in Yunnan is crucial, but remains limited to date (Lu et al. 2024b). This study reports two bioluminescent mushrooms in the Mycenaceae, based on a polyphasic approach: Mycena semivestipes (Peck) A.H. Sm., reported here for the first time for its bioluminescence and Roridomyces pruinosoviscidus (Corner) Blanco-Dios, a new geographical record for China. These findings contribute to our understanding of bioluminescent mushroom diversity from China, increasing the total number of known bioluminescent mushroom species in the region to 36. Additionally, the biological significance of bioluminescence is discussed. Materials and methods Sample collection, pure culture isolation and herbarium specimen preparation Fresh basidiomata that morphologically resemble Mycenaceae were collected from detached pieces of dead and rotting wood near a mountain stream in a wet forest in Yunnan Province, southwest China, in July 2023. All the important field information (altitude, colour, date, GPS coordinates, habitat and substrate) associated with the mushrooms was noted (Rathnayaka et al. 2024). The specimens were photographed in situ in daylight and in the laboratory using a Huawei P50 Pro camera (Shenzhen, P.R. China). The bioluminescent photographs were taken at night with a Canon EOS 80D camera (Tokyo, Japan) set to f/5.6, ISO 3200 and a shutter speed of 90 seconds in a black box. The basidiomata were then returned to the mycology laboratory at Qujing Normal University, where the specimens were completely dried in a hot air oven at 40℃ (Hu et al. 2022). The pure cultures were obtained using the spore print technique; the fresh mushroom caps were stuck on a sterile cover Petri plate with potato dextrose agar (PDA) and, after ten minutes, the mushroom caps with the Petri plate cover were removed; a new Petri plate cover was replaced and incubated for 24 h at 28℃. Germinated spores were transferred to a new PDA plate and incubated in an incubator (28℃) to observe and photograph bioluminescence and to facilitate further DNA 4Lu W et al extraction. All dried specimens were deposited in the Guizhou Medical University Herbarium (GMB) and living cultures were deposited in the Guizhou Medical University Culture Collection (GMBCC). Morphological study Macromorphological characteristics were described using the terminology of Largent (1986), based on notes from fresh collections and associated photographs. Kornerup and Wanscher’s colour terms and code were followed in this study (Kornerup and Wanscher 1967). The microscopic characteristics were described by using the concepts of Largent et al. (1977). Micromorphological observations were conducted using the methods described by Lu et al. (2024a). Freehand sections of the dried specimens were mounted in 5% potassium hydroxide (KOH) and stained with Congo red for microscopic examination. Melzer’s reagent was used to enhance contrast and assess the amyloid reaction in basidiospores. The light Eclipse 80i microscope (Olympus, Japan) was used to examine various features, including basidia, basidiospores, cystidia and hyphae. Measurements were conducted on at least 50 spores and their sizes were determined. Basidiospore dimensions were denoted as (a–) b–c (–d), where the range ‘b–c’ encompassed 90% or more of the measured values, with a and d representing the extreme values. Parameter Q refers to the interval of the length/width ratio of all basidiospores measured. Qm represented the mean Q value with standard deviation. Detailed illustrations of microstructures were sketched by hand using rehydrated materials and subsequently refined using Adobe Illustrator 2019. DNA extraction, PCR amplification and sequencing The total DNA was extracted from dried specimens and pure cultures using the Biospin Fungus Genomic DNA Extraction Kit-BSC14S1 (BioFlux, P.R. China), according to the manufacturer’s instructions, with minor modifications. The internal transcribed spacer region (ITS), large subunit (LSU) and small subunit (SSU) of the cistron coding for the ribosomal rRNAs and two protein-coding genes, translation elongation factor 1-α (tef1-α) and RNA polymerase II (rpb2), were amplified using the primer pairs ITS1/ITS4, LR0R/ LR5, NS1/NS4, 983F/2218R and 5F/7cR, respectively (Vilgalys and Hester 1990, White et al. 1990, Liu et al. 1999), in a total reaction volume of 25 μl containing 12.5 μl of 2x Master Mix (mixture of Easy Taq TM DNA Polymerase, dNTPs and optimised buffer (Beijing Trans Gen Biotech Co., Chaoyang District, Beijing, P.R. China), 8.5 μl distilled water, 2 μl DNA template and 1 μl of each primer. The cycle parameters were as follows: an initial denaturation at 94℃ for 5 min; denaturation at 94℃ for 30 s, annealing at 54℃ for 40 s (ITS, LSU, SSU and tef1-α) or at 58℃ for 90 s (rpb2) and an extension at 72℃ for 1 min for 35 cycles; with a final extension at 72℃ for 10 min; storage at 4℃. PCR products were sent to Sangon Biotech Co., Ltd. (Kunming, China) for sequencing with the PCR primers mentioned above. All newly-generated sequences in the present study were deposited in GenBank (https://www.ncbi.nlm.nih.gov/genbank, assessed on 25 September 2024). Hidden in the daylight: A polyphasic approach reveals two bioluminescent ... 5 Phylogenetic analyses Raw sequence reads (forward and reverse) were checked and manually edited for quality in BioEdit version 7.0.5 (Hall 1999) to correct base-calling errors, trim low-quality ends and confirm sequence orientation. Contigs were then assembled using Sequencher version 5.4.6 (Gene Codes Corporation 2016). The most similar sequences were identified using BLASTn (https://blast.ncbi.nlm.nih.gov/Blast.cgi, accessed on 10 June 2025) and were downloaded for phylogenetic analyses. Each dataset (ITS, LSU, SSU, tef 1-α and rpb2) was aligned using MAFFT version 7 on the online server platform (www.ebi.ac.uk/Tools/mafft; Katoh and Standley (2013)). The “auto” strategy was employed, allowing MAFFT to select the optimal alignment algorithm for each dataset. TrimAL version 1.2 (http://trimal.cgenomics.org, accessed 10 June 2025) was used to automatically remove gaps and ambiguous regions, applying the gt 0.4 algorithm. The final alignments for each locus were concatenated using SequenceMatrix v.1.7.8 to generate the multi-gene dataset. The final FASTA format was converted to PHYLIP and NEXUS formats using the Alignment Transformation Environment (ALTER) online programme (Glez-Peña et al. 2010). Prior to multi-locus analyses, single-gene tree topologies were inspected for topological conflicts at nodes with bootstrap support > 70%; none was observed. A concatenated dataset of ITS, LSU, SSU, tef1-α and rpb2 was assembled in BioEdit v.7.0.5, with gene regions concatenated in the order listed and partitioned by gene region, resulting in five character sets. Partitioned Maximum Likelihood (ML) analyses were conducted in RAxML-HPC2 v.8.2.12 (Stamatakis 2014) via the CIPRES Science Gateway v.3.3 (http://www.phylo.org/portal2, accessed on 13 November 2025) (Miller et al. 2010). A partitioned (mixed) model was implemented in RAxML using a partition file, with each gene region assigned its own GTRGAMMA substitution model. Node support was assessed with 1,000 rapid bootstrap replicates. Bayesian Inference (BI) analyses were conducted in MrBayes version 3.2.7a via the same web portal as the ML analyses (Ronquist et al. 2012). The analyses ran on XSEDE computational resources. Two independent runs of six Markov chains each for 20,000,000 generations, sampling every 200 generations (100,000 samples per chain) were undertaken. Preliminary inspection of an initial exploratory MCMC anlysis indicated that stationarity was not reached within the 2,000,000–5,000,000 generations; the final MCMC analysis was, therefore, run for 20,000,000 generations to achieve adequate mixing and convergence. Convergence and stationarity were assessed from both independent runs using three criteria: (1) the average standard deviation of split frequencies (ASDSF), which reached 0.009991 (threshold < 0.01); (2) effective sample sizes (ESS) of combined post-burn-in parameter traces, all of which exceeded 200, with most parameters showing ESS values well above 1,000, as confirmed in Tracer v.1.7.2 (Rambaut et al. 2018); and (3) log-likelihood and parameter trace plots exhibited stable, stationary “caterpillar”-like patterns with no observable trends and the two independent runs showed highly overlapping traces. Substitution models for each partition were selected under the Akaike Information Criterion (AIC) using MrModelTest v.2.2 (Nylander 2004). In these final runs, burn-in was assessed by visually inspecting the log-likelihood and parameter traces and all pre-stationary samples were removed. Examination of the 6Lu W et al Tracer plots indicated that stationarity was reached at ~ 1.0–1.2 million generations and we, therefore, conservatively discarded the first 10% of samples as burn-in. A 50% majority-rule consensus tree was constructed from the post–burn-in trees, retaining clades present in ≥ 50% of samples, with posterior probabilities provided for each clade. Phylogenetic trees were visualised using FigTree version 1.4.0 (Rambaut 2009) and were edited in Microsoft PowerPoint. The reliable bootstrap support values of ML (BS ≥ 70%) and Bayesian posterior probabilities (PP ≥ 0.90) were indicated above each branch. Taxon treatments Mycena semivestipes (Peck) A.H. Sm., 1947 • IndexFungorum 288544 Material a. kingdom: Fungi; phylum: Basidiomycota; class: Agaricomycetes; order: Agaricales; family: Mycenaceae; taxonRank: species; genus: Mycena; country: China; stateProvince: Yunnan; county: Chuxiong; verbatimElevation: 1871 m; verbatimLatitude: 24°54′31″ N; verbatimLongitude: 101°07′25″ E; year: 2023; month: July; day: 15; identifiedBy: Wenhua Lu; institutionID: Guizhou Medical University Herbarium (GMB); institutionCode: GMB1045; collectionCode: CX006; basisOfRecord: other material GMB1047, living culture GMBCC1066; GenBank accession numbers ITS: PQ373879 (GMB1045), PQ373880 (GMB1047), PQ373883 (GMBCC1066); LSU: PQ373885 (GMB1045), PQ373886 (GMB1047), PQ373889 (GMBCC1066); occurrenceID: 8DC70ECC-4DD5-5DDE-BE1A-B9A03672DE9E Description Basidiomata small. Pileus 3–15 mm diam., hemispheric, campaniform, expanding to flatten with age, initially convex to obtusely conical, margin white, centre greyishwhite (1B1) when young, surface wet, smooth, slimy, depressed, striate-plicate, translucid; centre light orange (5A4–5); margin white or translucid, thin, translucentstriate, hygrophanous in age; colour changes to greyish-orange (5B4–5), brownishorange (5C5–6) when mature. Context thin, fragile, translucent, white. Lamellae adnate to subdecurrent, whitish to white with 1–3 series of lamellulae, edges finely fimbriate under a lens, concolorous. Stipe 20–45 × 0.6–1.2 mm, cylindrical, hollow, brittle, brownish-orange to reddish-golden (6C7–8), milk-white (1A1–2) from base upwards, brown (6E7–8) at the base with whitish mycelium fascicles. Smell and taste none (Figs 1, 2). Bioluminescence: Only the caps and gills of basidiomata and the mycelia on the PDA emit yellowish-green light and the bioluminescence on spores was undetected (Fig. 3a). Hidden in the daylight: A polyphasic approach reveals two bioluminescent ... 7 Basidiospores (3.4–) 4.0–5.0 (–5.5) × (2.0–) 2.3–2.8 (–3.0) μm (n = 50), Q = 1.55−1.80, Qm = 1.7, ellipsoid to oblong, smooth, hyaline, containing oil droplets, thin-walled, inamyloid. Basidia 14.5–22 × 4–6.5 μm (n = 15), 4-sterigmata, clavate, some with oily contents, thin-walled. Cheilocystidia abundant, 15–35 × 7.5–13 μm, mainly clavate, cylindrical to fusiform, utriform with warty or finger-like protuberances, colourless, thin-walled. Pleurocystidia like cheilocystidia, 14–33.5 × 8–13 μm. Pileipellis a cutis with a well-developed cutis structure, hyphae 4–6 μm wide, cylindrical, parallel, with abundant, numerous sharp spines, colourless, thin-walled. Stipitipellis is a cutis with thin-walled hyphae. Caulocystidia clavate or fusiform. Clamp connections present on all hyphae and structures are abundant (Figs 1, 2). Distribution Canada, China, France, Sweden and the USA. Figure 1. Basidiomata of Mycena semivestipes (GMB1045). a, b, d Photographs in the daylight; c Stipe with white mycelium hairs (red arrow); e1 Photograph with the aid of a flashlight in the lab; e2 Bioluminescent photographs in complete darkness. Scale bars: a, b, d, e = 10 mm, c = 1 mm.  8Lu W et al Ecology Scattered, clustered or in groups on decaying trees in mixed coniferous and broadleaved forests in summer and autumn. Notes The ITS sequences of our Mycena collections had the highest BLAST score to that of M. semivestipes (LE-BIN 3362) with 98.62% (difference in three indels and three substitutions) and M. tintinnabulum (NSK 1017255) with 99% (difference in three indels and four substitutions). Morphologically, both M. semivestipes and M. tintinnabulum are very similar; the former differs in that the finger-like protuberances of cystidia are sparse, whereas in M. tintinnabulum, cheilocystidia bear numerous short finger-like or occasionally branched projections, giving a distinctly ornamented appearance (Smith 1937, Smith 1947, Na 2019). Mycena tintinnabulum is mainly found in Europe and the bioluminescence is present only in the mycelium. Our collections are consistent with M. semivestipes in many cases (small, lubricous basidiomata, adnate to subdecurrent lamellae, smooth amyloid spores, gelatinous pileipellis, cartilaginous context and cheilocystidia smooth or somewhat contorted and branched, but not numerous finger-like processes); Na (2019) reported that it is a new record for China, based on specimens from southwest China. Macromorphological characteristics indicate that the colour of the basidiomata in our Figure 2. Microscopic characteristics of Mycena semivestipes (GMB1045). aSpores; bBasidia; c Cheilocystidia; d Pleurocystidia; e Pileipellis hyphae; f Stipitipellis with caulocystidia. Scale bars = 10 µm.  Hidden in the daylight: A polyphasic approach reveals two bioluminescent ... 9 Acknowledgements SCK and ST 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. WL thanks Prof. Shi-Chen Shao and Xishuangbanna Primitive Forest Park for the sample collection. This work is partially supported by Chiang Mai University. The authors extend their appreciation to the Ongoing Research Funding Program (ORF-CTR-2025-6), King Saud University, Riyadh, Saudi Arabia. Author contributions Wenhua Lu and Alanoud T. Alfagham contributed equally to this work. References • Bäuerle J, Anke T, Jente R, Bosold F (1982) Antibiotics from basidiomycetes XVI. Antimicrobial and cytotoxic polyynes from Mycena viridimarginata karst. Archives of Microbiology 132: 194‑196. https://doi.org/10.1007/BF00508730 • Briones-Martin-Del-Campo M, Orta-Zavalza E, Juarez-Cepeda J, Gutierrez-Escobedo G, Cañas-Villamar I, Castaño I, De Las Peñas A (2014) The oxidative stress response of the opportunistic fungal pathogen Candida glabrata. Revista Iberoamericana de Micología 31: 67‑71. https://doi.org/10.1016/j.riam.2013.09.012 • Chew AC, Desjardin D, Tan Y, Musa MY, Sabaratnam V (2015) Bioluminescent fungi from Peninsular Malaysia—a taxonomic and phylogenetic overview. Fungal Diversity 70 (1): 149‑187. https://doi.org/10.1007/s13225-014-0302-9 • Chew AL, Tan YS, Desjardin DE, Musa MY, Sabaratnam V (2014) Four new bioluminescent taxa of Mycena sect. Calodontes from Peninsular Malaysia. Mycologia 106: 976‑988. https://doi.org/10.3852/13-274 • Cordero RB, Mattoon ER, Ramos Z, Casadevall A (2023) The hypothermic nature of fungi. Proceedings of the National Academy of Sciences 120 (19): e2221996120. https://doi.org/ 10.1073/pnas.2221996120 • Dauner LA, Karunarathna SC, Tibpromma S, Xu J, Mortimer PE (2021) Bioluminescent fungus Roridomyces viridiluminus sp. nov. and the first Chinese record of the genus Roridomyces, from Southwestern China. Phytotaxa 487: 233‑250. https://doi.org/ 10.11646/phytotaxa.487.3.4 • Desjardin DE, Oliveira AG, Stevani CV (2008) Fungi bioluminescence revisited. Photochemical & Photobiological Sciences 7: 170‑182. https://doi.org/10.1039/b713328f • Dutta AK, Paloi S, Hu Y, Axford S, Marciniak C (2020) Roridomyces phyllostachydis (Agaricales, Mycenaceae), a new bioluminescent fungus from Northeast India. Phytotaxa 459: 155‑167. https://doi.org/10.11646/phytotaxa.459.2.6 • Dybas CL (2019) Illuminating new biomedical discoveries: bioluminescent, biofluorescent species glow with promise. BioScience 69: 487‑495. https://doi.org/10.1093/biosci/biz052 16 Lu W et al • Emri T, Antal K, Varga K, Gila BC, Pócsi I (2024) The oxidative stress response highly depends on glucose and iron availability in aspergillus fumigatus. Journal of Fungi 10: 221. https://doi.org/10.3390/jof10030221 • Feng B, Yang ZL (2018) Studies on diversity of higher fungi in Yunnan, southwestern China: A review. Plant Diversity 40 (4): 165‑171. https://doi.org/10.1016/j.pld.2018.07.001 • Frankland JC (1998) Fungal succession-unravelling the unpredictable. Mycological Research 102: 1‑15. https://doi.org/10.1017/S0953756297005364 • Fukasawa Y, Osono T, Takeda H (2009) Effects of attack of saprobic fungi on twig litter decomposition by endophytic fungi. Ecological Research 24: 1067‑1073. https://doi.org/ 10.1007/s11284-009-0582-9 • Ge J, Lang X, Ji J, Qu C, Qiao H, Zhong J, Luo D, Hu J, Chen H, Wang S, Wang T, Li S, Li W, Zheng P, Xu J, Du H (2024) Integration of biological and information technologies to enhance plant autoluminescence. The Plant Cell 36 (11): 4703‑4715. https://doi.org/ 10.1093/plcell/koae236 • Gene Codes Corporation (2016) Sequencher [DNA sequence assembly software]. 5.4.6. Ann Arbor, MI, USA. URL: http://www.genecodes.com • Glez-Peña D, Gómez-Blanco D, Reboiro-Jato M, Fdez-Riverola F, Posada D (2010) ALTER: program-oriented conversion of DNA and protein alignments. Nucleic Acids Research 38: W14‑W18. https://doi.org/10.1093/nar/gkq321 • Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95‑98. • Heinzelmann R, Baggenstos H, Rudolf A (2024) Is the bioluminescence in many Mycena species overlooked? - A case study from M. crocata in Switzerland. Mycoscience MYC633 https://doi.org/10.47371/mycosci.2024.03.001 • Hu YW, Karunarathna SC, Li H, Galappaththi MC, Zhao CL, Kakumyan P, Mortimer PE (2022) The impact of drying temperature on basidiospore size. Diversity 14: 239. https:// doi.org/10.3390/d14040239 • Iqbal T, Rodrigues FA, Mahajan PV, Kerry JP (2009) Effect of time, temperature, and slicing on respiration rate of mushrooms. Journal of Food Science 74: 298‑303. https:// doi.org/10.1111/j.1750-3841.2009.01198.x • Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30: 772‑780. https://doi.org/10.1093/molbev/mst010 • Ke HM, Lu MR, Chang CC, Hsiao C, Ou JH, Taneyama Y, Tsai IJ (2023) Evolution and diversity of bioluminescent fungi. In: Pöggeler S, James T (Eds) Evolution of fungi and fungal-like organisms. Springer https://doi.org/10.1007/978-3-031-29199-9_12 • Kornerup A, Wanscher JH (1967) Methuen handbook of colour. Methuen & Co. Ltd • Kotlobay A, Sarkisyan K, Mokrushina Y, Marcet-Houben M, Serebrovskaya E, Markina N, Gonzalez Somermeyer L, Gorokhovatsky A, Vvedensky A, Purtov K, Petushkov V, Rodionova N, Chepurnyh T, Fakhranurova L, Guglya E, Ziganshin R, Tsarkova A, Kaskova Z, Shender V, Abakumov M, Abakumova T, Povolotskaya I, Eroshkin F, Zaraisky A, Mishin A, Dolgov S, Mitiouchkina T, Kopantzev E, Waldenmaier H, Oliveira A, Oba Y, Barsova E, Bogdanova E, Gabaldón T, Stevani C, Lukyanov S, Smirnov I, Gitelson J, Kondrashov F, Yampolsky I (2018) Genetically encodable bioluminescent system from fungi. Proceedings of the National Academy of Sciences 115 (50): 12728‑12732. https://doi.org/10.1073/pnas.1803615115 Hidden in the daylight: A polyphasic approach reveals two bioluminescent ... 17 • Largent DL, Johnson D, Watling R (1977) How to identify mushrooms to genus III: microscopic features. Mad River Press • Largent DL (1986) How to Identify Mushrooms to Genus I-Macroscopic Features. Revised, Subsequent, 1. Mad River Pr Inc, Eureka, California, 425‑431 pp. • Liu YJ, Whelen S, Hall BD (1999) Phylogenetic relationships among ascomycetes: evidence from an RNA polymerase II subunit. Molecular Biology and Evolution 16 (12): 1799‑1808. https://doi.org/10.1093/oxfordjournals.molbev.a026092 • Lu W, Suwannarach N, Lumyong S, Elgorban A, Dai D, Dutta A, Han L, Tibpromma S, Karunarathna S (2024a) Molecular phylogeny and morphology reveal two new species of Conocybe (Bolbitiaceae, Agaricales) from southwest China. New Zealand Journal of Botany 62: 501‑518. https://doi.org/10.1080/0028825x.2024.2327117 • Lu W, Priyashantha AK, Galappaththi MC, Tibpromma S, Dai DQ, Patabendige NM, Premarathne BM, Kulasuriya DM, Ediriweera AN, Nimalrathna TS, Suwannarach N, Lumyong S, Tang A, Shao SC, Karunarathna SC (2024b) Fungal bioluminescence: Past, present, and future. Diversity 16: 539. https://doi.org/10.3390/d16090539 • Mihail JD, Bilyeu L, Lalk S (2018) Bioluminescence expression during the transition from mycelium to mushroom in three North American Armillaria and Desarmillaria species. Fungal Biology 122: 1064‑1068. https://doi.org/10.1016/j.funbio.2018.08.007 • Miller MA, Pfeiffer W, Schwartz T (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Proceedings of the Gateway Computing Environments Workshop (GCE), New Orleans, November. LA, 1-8 pp. https://doi.org/ 10.1109/GCE.2010.5676129 • Mitiouchkina T, Mishin AS, Somermeyer LG, Markina NM, Chepurnyh TV, Guglya EB, Karataeva TA, Palkina KA, Shakhova ES, Fakhranurova LI (2020) Plants with genetically encoded autoluminescence. Nature Biotechnology 38: 944‑946. https://doi.org/10.1038/ s41587-020-0500-9 • Nagamune K, Hosaka K, Kigawa S, Sugawara R, Sotome K, Nakagiri A, Endo N (2024) Two new Mycena section Calodontes species: One newly discovered and the other new to Japan. Mycoscience 65: 111‑122. https://doi.org/10.47371/mycosci.2024.02.002 • Na Q (2019) Taxonomy and phylogeny of Mycena in China. Jilin Agricultural University, Jilin. [In Chinese]. URL: https://link.cnki.net/doi/10.27163/d.cnki.gjlnu.2019.000016doi: 10.27163/d.cnki.gjlnu.2019.000016 • Nimalrathna TS, Tibpromma S, Nakamura A, Galappaththi MC, Xu J, Mortimer PE, Karunarathna SC (2022) The case of the missing mushroom: A novel bioluminescent species discovered within Favolaschia in southwestern China. Phytotaxa 539: 244‑256. https://doi.org/10.11646/phytotaxa.539.3.3 • Nylander J (2004) MrModeltest V2. Program distributed by the author. Bioinformatics 24: 581‑583. https://doi.org/10.1093/bioinformatics/btm388 • Oliveira AG, Stevani CV, Waldenmaier HE, Viviani V, Emerson JM, Loros JJ, Dunlap JC (2015) Circadian control sheds light on fungal bioluminescence. Current Biology 25: 964‑968. https://doi.org/10.1016/j.cub.2015.02.021 • Perry BA, Desjardin DE, Stevani CV (2025) Diversity, distribution, and evolution of bioluminescent fungi. Journal of Fungi 11 (1): 19. https://doi.org/10.3390/jof11010019 • Qiang TT, Bai LC (2023) Mycena cinereoalba sp. nov. from Qinghai province, China. Mycotaxon 137: 803‑811. https://doi.org/10.5248/137.803 • Rambaut A (2009) FigTree v1. 3.1. http://tree.bio.ed.ac.uk/software/fgtree/.2009 18 Lu W et al • Rambaut A, Drummond AJ, Xie D, Baele G, Suchard MA (2018) Posterior Summarization in Bayesian Phylogenetics Using Tracer 1.7. Systematic Biology 67 (5): 901‑904. https:// doi.org/10.1093/sysbio/syy032 • Rathnayaka AR, Tennakoon DS, Jones GE, Wanasinghe DN, Bhat DJ, Priyashantha AK, Stephenson SL, Tibpromma S, Karunarathna SC (2024) Significance of precise documentation of hosts and geospatial data of fungal collections, with an emphasis on plant-associated fungi. New Zealand Journal of Botany1‑28. https://doi.org/ 10.1080/0028825X.2024.2381734 • Rexer KH (1994) Die Gattung Mycena sl: Studien zu ihrer Anatomie, Morphologie und Systematik. Uitgever niet vastgesteld • Ronquist F, Teslenko M, van der Mark P, Ayres D, Darling A, Höhna S, Larget B, Liu L, Suchard M, Huelsenbeck J (2012) MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space. Systematic Biology 61 (3): 539‑542. https://doi.org/10.1093/sysbio/sys029 • Seok SJ, Jin YJ, Yoo KB, Hong SB, Kwon SW, Kim SJ, Kim JS (2015) Notes on the new species of genus Mycena in Korea. The Korean Journal of Mycology 43: 224‑230. https:// doi.org/10.4489/KJM.2015.43.4.224 • Smith AH (1937) Studies in the Genus Mycena IV. Mycologia 29: 338‑354. https://doi.org/ 10.1080/00275514.1937.12017212 • Smith AH (1947) North American species of Mycena. University of Michigan Press, Ann Arbor, Michigan, 687 pp. • Smithsonian (2024) Bioluminescence first evolved in animals at least 540 million years ago. https://www.sciencedaily.com/releases/2024/04/240423204148.htm. Accessed on: 2024-10-07. • Soares CC, Cabral TS, Vargas-isla R, Cardoso JS, Rodrigues DP, Ishikawa NK, Oliveira JJ (2024) Mycena lamprocephala, a new luminescent species from the Brazilian Amazon. Phytotaxa 634: 187‑203. https://doi.org/10.11646/phytotaxa.634.3.1 • Stamatakis A (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30: 1312‑1313. https://doi.org/10.1093/bioinformatics/ btu033 • Syed AJ, Anderson JC (2021) Applications of bioluminescence in biotechnology and beyond. Chemical Society Reviews 50: 5668‑5705. https://doi.org/10.1039/D0CS01492C • Tisch D, Schmoll M (2010) Light regulation of metabolic pathways in fungi. Applied Microbiology and Biotechnology 85: 1259‑1277. https://doi.org/10.1007/ s00253-009-2320-1 • Vilgalys R, Hester M (1990) Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238‑4246. https://doi.org/10.1128/jb.172.8.4238-4246.1990 • Warris A, Ballou ER (2019) Oxidative responses and fungal infection biology. Seminars in Cell & Developmental Biology 89: 34‑46. https://doi.org/10.1016/j.semcdb.2018.03.004 • Weinstein P, Delean S, Wood T, Austin AD (2016) Bioluminescence in the ghost fungus Omphalotus nidiformis does not attract potential spore-dispersing insects. IMA Fungus 7: 229‑234. https://doi.org/10.5598/imafungus.2016.07.02.01 • Wei R, Ge Y, Qi L, Han M, Zeng H, Hu Y, Zou L, Cheng X, Wu X, Na Q (2024) Revealing brownish Mycena diversity in China: New discoveries and taxonomic insights. Journal of Fungi 10: 439. https://doi.org/10.3390/jof10060439 Hidden in the daylight: A polyphasic approach reveals two bioluminescent ... 19 • White TJ, Bruns TD, Lee SB, Taylor JW (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, JJ S, White TJ (Eds) PCR Protocols: A guide to methods and applications. Academic Press, Inc., New York, 315–322 pp. https://doi.org/10.1016/B978-0-12-372180-8.50042-1 • Yaakoub H, Mina S, Calenda A, Bouchara JP, Papon N (2022) Oxidative stress response pathways in fungi. Cellular and Molecular Life Sciences 79: 333. https://doi.org/10.1007/ s00018-022-04353-8 • Zhang Y, Mo M, Yang L, Mi F, Cao Y, Liu C, Tang X, Wang P, Xu J (2021) Exploring the species diversity of edible mushrooms in Yunnan, Southwestern China, by DNA Barcoding. Journal of Fungi 7: 310. https://doi.org/10.3390/jof7040310 • Zheng P, Ge J, Ji J, Zhong J, Chen H, Luo D, Li W, Bi B, Ma Y, Tong W, Han L, Ma S, Zhang Y, Wu J, Zhao Y, Pan R, Fan P, Lu M, Du H (2023) Metabolic engineering and mechanical investigation of enhanced plant autoluminescence. Plant Biotechnology Journal 21: 1671‑1681. https://doi.org/10.1111/pbi.14068 20 Lu W et al