Three New Mushroom Species Uncovered in China’s Forests Through DNA and Morphology

Chinese mycologists have described three previously unknown species of the mushroom genus Hohenbuehelia, a group of small, fan-shaped fungi that had been comparatively neglected in the country’s forests. The new species, named Hohenbuehelia albomarginata, H. guizhouensis and H. striata, were identified from specimens collected in Yunnan, Guizhou, Sichuan and Hebei provinces, and their discovery brings the number of Hohenbuehelia species known from China to seventeen. The finding, published in the journal MycoKeys, rests on a combination of detailed microscopic anatomy and a three-gene DNA analysis that places each of the new fungi on its own strongly supported branch of the evolutionary tree.

Hohenbuehelia is a cosmopolitan genus in the family Pleurotaceae, the same family that contains the familiar oyster mushroom. The genus was validly established by Schulzer in 1866 and currently comprises roughly 202 valid names worldwide, with more than 50 species generally accepted as distinct. Species in the group are characterized by small fruiting bodies with spathulate, fan-shaped or half-circular caps, gills that are decurrent or radiate from the point of attachment, a reduced or absent stipe, ellipsoid spores and distinctive thick-walled metuloid pleurocystidia, the sterile cells embedded in the gill tissue. Historically the genus was shuffled into Pleurotus because of the similar cap shape, but phylogenetic analyses based on the internal transcribed spacer region and the large subunit ribosomal RNA gene, together with the presence of a gelatinous layer between the flesh and the cap cuticle, confirmed Hohenbuehelia as a monophyletic genus of its own within the Pleurotaceae.

The genus matters beyond taxonomy. A few species, such as H. reniformis and H. petaloides, have been reported as edible mushrooms in China. Several species are sources of antioxidant properties as well as antitumor and antiviral activities, and some are predatory: they attack nematodes by producing adhesive knobs on their hyphae or on germinated asexual conidia, a trait that has long interested researchers exploring biological control of nematodes. The diversity of bioactive compounds found in Hohenbuehelia species and their potential applications underline the importance of detailed taxonomic study of the genus, since precise species limits are the foundation on which any such applied work depends.

The specimens behind the new species were collected between 2002 and 2024, from subtropical broad-leaved forests in Yunnan, Guizhou and Sichuan and warm-temperate deciduous broad-leaved forests in Hebei. Several of the collections were made on wood in forests with trees of the beech family, Fagaceae, at altitudes ranging from roughly 1,470 to 3,240 meters. All nine specimens examined in the study are deposited in the Herbarium of Cryptogams at the Kunming Institute of Botany, Chinese Academy of Sciences. Genomic DNA was extracted from silica-gel dried or herbarium material using the CTAB method, and three regions were amplified and sequenced: the internal transcribed spacer, the nuclear large subunit ribosomal RNA gene, and the translation elongation factor 1-alpha gene, a standard trio for resolving species boundaries in mushrooms.

The phylogenetic analysis assembled a three-gene matrix of 87 samples and 2,242 aligned bases per sample, combining 22 newly generated sequences with data retrieved from GenBank, including sequences from type, holotype and epitype specimens of related species. Four Pleurotus species served as outgroups. The team used Maximum Likelihood and Bayesian Inference analyses, with substitution models selected by PartitionFinder under the Bayesian Information Criterion; the best-fitting models were GTR+GAMMA for the ITS dataset and GTR+GAMMA+I for nrLSU and TEF1-alpha. Bootstrap support was calculated from 1,000 replicates, and Bayesian runs were continued until the standard deviation of split frequencies fell below 0.01 and effective sample sizes exceeded 200. Because the tree topologies from the two approaches were nearly identical, differing only slightly in statistical support, the Maximum Likelihood tree was used as the reference figure.

The result was unambiguous for species delimitation. Each of the three new species formed a strongly supported monophyletic clade, with Maximum Likelihood bootstrap values of 100 percent and Bayesian posterior probabilities of 1.0 for each. The relationships between these species and their closest allies received more variable support, ranging from 73 to 100 percent, which the authors note indicates that the deeper internodal relationships within Hohenbuehelia may require additional molecular markers and broader taxon sampling to be fully resolved. In the tree, H. albomarginata sits in a clade related to H. fluxilis, H. josserandii, H. reniformis, H. atrocoerulea and H. canadensis; H. striata clusters with H. thornii, H. petaloides, H. wilhelmii and H. angustata; and H. guizhouensis emerges as a sister lineage to the Costa Rican species H. carlothornii, with that pairing supported by a bootstrap value of 98 percent.

Each new species also carries a distinctive anatomy. Hohenbuehelia albomarginata, whose name refers to the white margin of its cap, produces small gray-brown to brown fruiting bodies 0.5 to 2.5 centimeters wide, frosted with white tomentum and bearing decurrent, crowded gills that yellow with age, attached by a short, stout pseudostipe. Its basidiospores measure 5.5 to 8 micrometers long and 3 to 4.5 micrometers wide, with mean dimensions of 6.6 by 3.7 micrometers, and are mostly elongate. Its thick-walled pleurocystidia, 45 to 55 by 10 to 14 micrometers, are sometimes partly covered with refringent crystalline granules. The species is known from Yunnan, Sichuan and Hebei, fruiting on wood from summer to autumn, and can be separated from lookalikes such as H. josserandii, which has a darker pileus, sparser lamellae and slightly longer spores of 7.5 to 9 by 3.5 to 4 micrometers, and H. reniformis, which has cylindrical spores.

Hohenbuehelia guizhouensis, named for Guizhou Province where the type specimen was collected in 2018 at an altitude of 1,820 meters, lacks a pseudostipe entirely, with gills radiating from the point of attachment, and produces mostly elongate, sometimes ellipsoid to subreniform spores of 6 to 8 by 3 to 4 micrometers, with a mean length-to-width ratio of 2. Although it is phylogenetically close to H. carlothornii, the two differ in spore shape and in ecology: the new species fruits on fallen wood in subtropical broad-leaved forests in southwest China, whereas H. carlothornii was isolated from soil and litter and fruits on dead angiosperm wood in tropical wet forest and premontane rain forest in Costa Rica. Hohenbuehelia striata, the largest of the three, has caps 4.5 to 6 centimeters wide that are brown to yellow-brown, glabrous, shiny and very faintly translucent-striate near the margin, the feature that gives the species its name. Its spores are mostly ellipsoid, 5.5 to 6.8 micrometers long with means of 6.3 by 4.6 micrometers, distinguishing it from H. angustata, which has smaller spores, and from H. thornii, which has cylindrical spores and a small off-white pileus. It is known only from Yunnan, fruiting on wood in summer.

The authors emphasize that only a few morphological characters have traditionally been available for classifying Hohenbuehelia, and that the color of the fruiting body varies across a broad range, which is precisely why molecular methods have become essential for accurate classification and identification. Recent years have seen multi-locus phylogenetic approaches accelerate the description of new species across the genus, from Europe and North and South America to Central Asia, Russia and Southeast Asia. In China, species have now been recorded in provinces including Jilin, Inner Mongolia, Shanxi, Hunan, Guangdong and Hainan, with H. petaloides and H. reniformis the most commonly recorded taxa both by geographic spread and by the total number of finds recorded under those names.

The study also underscores the persistent gaps in knowledge. Southwestern China, where the three new species were collected, is widely recognized as a global biodiversity hotspot with high levels of microbial diversity, yet the authors stress that species diversity of Hohenbuehelia in the country remains poorly understood. They argue that broader taxon sampling combined with molecular and morphological data is needed to fully capture the genus’s richness, and that future surveys should target understudied regions and habitats where additional species, including cryptic ones, are likely waiting. Given the genus’s documented roles in nematode predation and its repertoire of bioactive properties, each new species added to the catalogue is also a potential new lead for applied research, provided the taxonomy keeps pace with the exploration.

Subject of Research: Morphological and multi-locus phylogenetic description of three new Hohenbuehelia mushroom species from Chinese forests

Article Title: Three new species of Hohenbuehelia ( Agaricales , Basidiomycota ) from China based on morphological and multi-locus phylogenetic evidence

Article References: Luo, T., Che, N.-J., Feng, W.-C., Cui, Y.-Y., & Li, J. (2026). Three new species of Hohenbuehelia (Agaricales, Basidiomycota) from China based on morphological and multi-locus phylogenetic evidence. MycoKeys, 142, 119-137. https://doi.org/10.3897/mycokeys.142.204939

Image Credits: AI Generated

DOI: 10.3897/mycokeys.142.204939

Keywords: Hohenbuehelia, Agaricales, Pleurotaceae, new species, China, phylogenetics, ITS, nrLSU, TEF1-alpha, taxonomy, mycology, biodiversity