Puffballs, the common name for members of the family Lycoperdaceae, are among the most familiar gasteroid fungi, producing spherical, pear-shaped, or top-shaped fruiting bodies in which spores mature internally rather than on exposed gills. The family was established in 1820 with the type genus Lycoperdon and has long fascinated mycologists because its members play significant roles in decomposing organic matter and even in accumulating heavy metals in soils. Some species, such as certain Calvatia, have been used as food and in traditional medicine for their hemostatic properties. Yet despite more than 1,400 recorded names, only around 430 species are currently accepted, and the true diversity of the group is believed to be far greater.

The story of the new genus began during fieldwork in the Wumengshan National Nature Reserve in Daguan County, Zhaotong, Yunnan Province, at an elevation of roughly 1,882 meters, where fruiting bodies were found growing on the ground in a forest of the bamboo Chimonobambusa tumidissinoda. Fresh specimens were photographed in situ, dried at 40 degrees Celsius, and deposited in the herbarium of Southwest Forestry University in Kunming. The mature fruiting bodies are small, measuring only 8 to 15 millimeters across and 9 to 14 millimeters tall, with a densely spiny outer peridium that is white when fresh and cream to pinkish-buff when dry.

What truly sets Longipediperdon spineum apart is visible only under the microscope. Its basidiospores are ovoid to oblong, colorless, and thick-walled, measuring about 4.4 to 5.8 micrometers long, and each carries a remarkably long pedicel, a stalk-like appendage ranging from 4 to 15.5 micrometers. This elongated spore stalk is the feature that inspired the genus name, which combines Latin roots meaning long, foot, and puffball. The spore surfaces bear spines shorter than one micrometer, and the basidia, the spore-producing cells, are bubble-shaped to subcylindrical with four sterigmata and a simple septum at the base.

To test whether these morphological quirks reflected genuine evolutionary separation, the team extracted genomic DNA from dried specimens using a CTAB-based kit and amplified three molecular markers: the internal transcribed spacer region (ITS), the nuclear large subunit ribosomal RNA gene (nLSU), and the RNA polymerase II subunit 2 gene (rpb2). These loci are the workhorses of fungal phylogenetics, offering a balance of variability and reliability that allows closely related species to be distinguished. Sequences were aligned with MAFFT, manually adjusted in AliView, and concatenated into a combined dataset.

The phylogenetic analysis was ambitious in scale. The combined ITS, nLSU, and rpb2 matrix included sequences from 271 fungal specimens representing 69 species, with a total alignment length of 2,455 base pairs and 595 distinct alignment patterns. Maximum likelihood trees were inferred with IQ-TREE using 5,000 ultrafast bootstrap replicates and 1,000 SH-aLRT tests, while Bayesian inference was run through the CIPRES Science Gateway with two independent runs of half a million generations each. Mycenastrum corium served as the outgroup to root the tree within Lycoperdaceae.

The result was unambiguous. Longipediperdon spineum formed an independent monophyletic lineage nested firmly within Lycoperdaceae, distinct from all 29 previously recognized genera in the family. Branch support met the stringent thresholds of maximum likelihood bootstrap values of at least 70 percent and Bayesian posterior probabilities of at least 0.95. In other words, the DNA evidence and the morphology told the same story: this was not merely an odd-looking Lycoperdon but a lineage that had been traveling its own evolutionary path.

Morphological comparisons reinforced the genetic verdict. The new genus most closely resembles Lycoperdon pulcherrimum in its subglobose to obpyriform fruiting bodies and Lycoperdon-type capillitium, the sterile thread network that helps disperse spores. However, L. pulcherrimum produces much larger fruiting bodies, 20 to 50 millimeters in diameter, and globose spores, whereas the new species is smaller and has elongated, pedicellate spores. The authors also distinguished Longipediperdon from 17 other genera, including Bovista, Calvatia, Morganella, and the recently described Lycoperdia and Tortoperdon, using characters such as capillitium type, spore ornamentation, and peridial structure. Notably, Morganella lacks capillitium altogether, while Longipediperdon possesses the Lycoperdon-type, occasionally branched, thick-walled threads.

The discovery carries broader implications for fungal biodiversity. Estimates suggest that only about 155,000 of an estimated 2.5 million fungal species have been formally described, and the Agaricomycetes, the class to which puffballs belong, remain poorly documented across their nearly global range. Recent multilocus studies have estimated that the major lineages of Lycoperdaceae diverged between roughly 26.7 and 75.5 million years ago, supporting the recognition of 19 genera, a number that has since climbed to 29 with integrative taxonomic work. Each new genus, such as Longipediperdon, refines our understanding of how this ecologically important family diversified.

For now, Longipediperdon spineum is known only from its type locality and a nearby paratype collection in Yunnan, both gathered in July 2025. The authors emphasize that finding a new genus within a well-studied family demonstrates that considerable undescribed diversity still hides in forest floor litter, particularly in biodiversity hotspots like southern China. Future work, they argue, should prioritize comprehensive sampling across the family’s range to map its global distribution patterns and evolutionary history, and perhaps to uncover more long-legged spores waiting in the leaf litter.

Subject of Research: Phylogeny and taxonomy of the puffball family Lycoperdaceae and the description of a new genus from China

Article Title: Phylogeny and taxonomy of Lycoperdaceae : Longipediperdon gen. nov., a new genus from China based on molecular and morphological evidence

Article References: Yang, X., Duan, S., Li, S., Gao, X., Yang, R., Jiang, D., Zhou, H., Zhao, C., & Shen, S. (2026). Phylogeny and taxonomy of Lycoperdaceae: Longipediperdon gen. nov., a new genus from China based on molecular and morphological evidence. MycoKeys, 136, 323-340. https://doi.org/10.3897/mycokeys.136.188385

Image Credits: AI Generated

DOI: 10.3897/mycokeys.136.188385

Keywords: Lycoperdaceae, puffballs, Longipediperdon, new genus, mycology, phylogenetics, ITS, nLSU, rpb2, China, taxonomy, fungal biodiversity