Fossil Fungi Research Articles

Maastrichtian palaeoenvironments and palaeoclimate reconstruction in southern South America (Patagonia, Argentina) based on fossil fungi and algae using open data resources

ABSTRACT The use of non-pollen palynomorphs (NPP), particularly fossil fungi and algae, as palaeobiological proxies for Late Cretaceous palaeoenvironmental and palaeoclimatic reconstructions of warm-to-hot greenhouse conditions, can enhance our understanding of climate change impacts on modern Patagonian environments. This study aimed to reconstruct the Maastrichtian palaeoenvironment and palaeoclimate in the Cañadón Asfalto Basin (CAB, Chubut Province) by testing these NPPs as proxies using the Nearest Living Relative method (NLR). Moreover, using modern ecological requirements from open-source databases, such as GBIF and processing it with an open-source, cross-platform tool like QGIS, linked with Köppen-Geiger shapefiles, provided evidence of climate-driven palaeo-distribution patterns of fungal and algal diversity at CAB. Applying modern ecological requirements and biogeographic distribution data, we reconstructed the palaeoclimate as temperate with evenly distributed precipitation and warm summers, corresponding to the Cfb climate zone in Köppen-Geiger classifications. Additionally, our methodology produced reliable results regarding Cenozoic floras’ physiognomies based on fossil fungi, revealing a transition from sparsely wooded areas with palms and prairies to complex forest ecosystems with palms, deciduous trees, and shrubland. Furthermore, testing Cretaceous algae with the NLR method, for the first time, provided comprehensive insights into past water body characteristics, including trophic state and water quality.

Classes and phyla of the kingdom Fungi

AbstractFungi are one of the most diverse groups of organisms with an estimated number of species in the range of 2–3 million. The higher-level ranking of fungi has been discussed in the framework of molecular phylogenetics since Hibbett et al., and the definition and the higher ranks (e.g., phyla) of the ‘true fungi’ have been revised in several subsequent publications. Rapid accumulation of novel genomic data and the advancements in phylogenetics now facilitate a robust and precise foundation for the higher-level classification within the kingdom. This study provides an updated classification of the kingdom Fungi, drawing upon a comprehensive phylogenomic analysis of Holomycota, with which we outline well-supported nodes of the fungal tree and explore more contentious groupings. We accept 19 phyla of Fungi, viz. Aphelidiomycota, Ascomycota, Basidiobolomycota, Basidiomycota, Blastocladiomycota, Calcarisporiellomycota, Chytridiomycota, Entomophthoromycota, Entorrhizomycota, Glomeromycota, Kickxellomycota, Monoblepharomycota, Mortierellomycota, Mucoromycota, Neocallimastigomycota, Olpidiomycota, Rozellomycota, Sanchytriomycota, and Zoopagomycota. In the phylogenies, Caulochytriomycota resides in Chytridiomycota; thus, the former is regarded as a synonym of the latter, while Caulochytriomycetes is viewed as a class in Chytridiomycota. We provide a description of each phylum followed by its classes. A new subphylum, Sanchytriomycotina Karpov is introduced as the only subphylum in Sanchytriomycota. The subclass Pneumocystomycetidae Kirk et al. in Pneumocystomycetes, Ascomycota is invalid and thus validated. Placements of fossil fungi in phyla and classes are also discussed, providing examples.

A new fossil species of Meliolinites Selkirk associated with Rhodoleia leaves from the Upper Pliocene of southwestern China

ABSTRACT Fossil epifoliar fungi are valuable indicators of paleoenvironment and paleoecology. The Meliolaceae, members of which typically inhabit the surface of living plants as biotrophs or pathogens, is one of the largest groups of epifoliar fungi. In this study, we report a novel fossil species of Meliolinites Selkirk (fossil Meliolaceae), Meliolinites tengchongensis, on the lower epidermis of compressed fossil Rhodoleia (Hamamelidaceae) leaves from the Upper Pliocene Mangbang Formation of Tengchong, Yunnan, southwestern China. Meliolinites tengchongensis is characterized by web-like, superficial, brown to dark brown, septate, and branching mycelia bearing 2-celled appressoria and unicellular phialides. The fungal colonies also include ellipsoidal, 5-celled, 4-septate ascospores and dark brown perithecia with suborbicular outline and verrucose surface. The well-preserved vegetative and reproductive organs help us to explore the potential disease process of the new fossil species. Besides, the presence of fungal remains indicates that the fungal taxon might have maintained its host preference since at least the Late Pliocene. Furthermore, the occurrence of both fossil fungi and their host plants in Tengchong indicate a subtropical–tropical, warm, and humid climate during the Late Pliocene, whereas the distribution pattern of the fungi on the host leaves suggests that Rhodoleia may have been a part of the middle–upper canopies in the Tengchong Late Pliocene multilayered forest.

Summer-Wet Hydrologic Cycle during the Middle Miocene of the United States: New Evidence from Fossil Fungi.

Hydrologic reconstructions from North America are largely unknown for the Middle Miocene. Examination of fungal palynomorph assemblages coupled with traditional plant-based palynology permits delineation of local, as opposed to regional, climate signals and provides a baseline for study of ancient fungas. Here, the Fungi in a Warmer World project presents paleoecology and paleoclimatology of 351 fungal morphotypes from 3 sites in the United States: the Clarkia Konservat-Lagerstätte site (Idaho), the Alum Bluff site (Florida), and the Bouie River site (Mississippi). Of these, 83 fungi are identified as extant taxa and 41 are newly reported from the Miocene. Combining new plant-based paleoclimatic reconstructions with funga-based paleoclimate reconstructions, we demonstrate cooling and hydrologic changes from the Miocene climate optimum to the Serravallian. In the southeastern United States, this is comparable to that reconstructed with pollen and paleobotany alone. In the northwestern United States, cooling is greater than indicated by other reconstructions and hydrology shifts seasonally, from no dry season to a dry summer season. Our results demonstrate the utility of fossil fungi as paleoecologic and paleoclimatic proxies and that warmer than modern geological time intervals do not match the "wet gets wetter, dry gets drier" paradigm. Instead, both plants and fungi show an invigorated hydrological cycle across mid-latitude North America.

Triplinerved cinnamon from the Siwalik (middle Miocene) of eastern Himalaya: Systematics, epifoliar fossil fungi, palaeoecology and biogeography

Cinnamomum Schaeffer (avocado, family Lauraceae), commonly known as cinnamon tree, is a highly diverse, economically important evergreen element of tropical and subtropical regions of the world. In the present work, several compressed leaf remains similar to modern leaves of Cinnamomum are recovered from the lower part of the Siwalik strata (middle Miocene) of Darjeeling foothills, eastern Himalaya. The fossil specimens are characterized by an elliptic to ovate lamina, acute to short acuminate apex, round to acute base, basal as well as suprabasal acrodromous type of primary venation, and anomocytic type of stomata. Based on leaf architecture (macro- and micromorphological features) and using leaf epidermal anatomy, these Siwalik fossil leaves are confidently assigned to the genus Cinnamomum and are recognized as a new species: C. miocenicum Mahato, Hazra et Khan. The evidence of current Siwalik specimens and earlier-reported fossil species similar to thermophilic Cinnamomum in appreciable numbers from other Siwalik localities (Darjeeling, Uttarakhand, and Nepal) suggests that Cinnamomum was a common Siwalik element, pointing toward the existence of tropical, warm and humid climate conditions during the Siwalik sedimentation time. This conclusion is also supported by epifoliar fungal remains recovered from cuticular fragments of this lauraceous taxon. This finding also represents an essential source of data for understanding Cinnamomum’s evolution and deep time diversification. We review in detail the biogeographic history and suggest possible migratory routes of the genus from an Asian perspective.

Preservation, diagenetic transformation and paleoecological significance of fungal saccharides from lignites and fossil wood

Fungi are a common component of terrestrial ecosystems, although their preservation as molecular fossils is relatively rare. Saccharides, such as arabitol, mannitol and trehalose, are important in biochemical processes, and are the dominant compounds in fungi, lichens and yeasts. Here we present gas chromatography–mass spectrometry (GC–MS) analysis of fungal saccharides in brown coal extracts and compare their distribution to extant white, brown, and soft-rot fungi. Distribution differs depending on the type of decay in modern and fossil wood-decomposing fungi. An arabitol/(arabitol + trehalose) ratio higher than 0.6 for extant and fossil fungi is diagnostic for soft and brown rot, while values below 0.4 are characteristic of white rot fungi. Values between 0.4 and 0.6 suggest a mixed origin of fungal saccharides. The observed differences may shed light on the type of fungal activity, in particular relatated to climatic conditions of the Cenozoic.We conclude that saccharides can be good indicators, sensitive to climate change, of wood degradation by fungi in thermally immature organic matter. As a case study, early Cenozoic lignites were formed on low latitudes in the tropics during or near the Paleocene/Eocene thermal maximum, as extant soft rot fungi can withstand extremes of temperature and humidity. The presence of these fungi is confirmed by the dominance of arabitol over trehalose in most of the Lower Cenozoic lignites and arabitol/(arabitol + trehalose) ratio values higher than 0.6. In contrast, Miocene detritic coals, which were formed in temperate to subtropical climates, contain saccharides derived from a mixture of different fungi with white rot predominance.

The core of Sporocarpon asteroides, an enigmatic fungal fossil from the Carboniferous

The core of Sporocarpon asteroides, an enigmatic fungal fossil from the Carboniferous

Szaferomyces pliocenicus nov. gen., nov. sp. from the Upper Pliocene deposits of Mizerna (Poland), a fossil fungus showing close resemblance to modern powdery mildews

A fungal chasmothecium-like structure resembling those of the contemporary members of the Erysiphaceae was found during palynological investigations of Upper Pliocene deposits from Mizerna (borehole Mizerna-Nowa), southern Poland. This chasmothecium-like remain, preserved without any asci, ascospores and mycelium attached, is morphologically most similar to the chasmothecia of modern powdery mildews from the genera Erysiphe, Neoerysiphe, Podosphaera, and Salmonomyces. It differs from hitherto described fossil or modern fungi producing globose, closed ascomata with appendage-like setae. Therefore, a new fossil-genus and species name, Szaferomyces pliocenicus, is introduced for the fossil remains from Mizerna. Contrary to all previous putative fossils of powdery mildews, Szaferomyces pliocenicus nov. gen., nov. sp. is the most probable fossil of spore-producing organs of the Erysiphaceae found so far. However, due to the absence of some diagnostic morphological structures (other than presumptive chasmothecium) and DNA data, the affinity of this fungal fossil remains unclear.

Fungal-induced fossil biomineralization

Exceptional preservation of fossils has often been attributed to the actions of bacteria that aid in the preservation of soft tissues that normally decay rapidly. However, it is well known that fungi play a major role in organic matter decomposition, biogeochemical cycling of elements, and metal-mineral transformations in modern ecosystems. Although the fungal fossil record can be traced back over a billion years, there are only a few recorded examples of fungal roles in fossilization. In this research, we have carried out a detailed geobiological investigation on early Pleistocene hyena coprolites (fossilized dung) in an attempt to ascertain possible fungal involvement in their formation. Using an advanced microscopic and mineralogical approach, we found that numerous hydroxyapatite nanofibers (25-34nm on average), interwoven to form spheroidal structures, constituted the matrix of the coprolites in addition to food remains. These structures were found to be extremely similar in texture and mineral composition to biominerals produced during laboratory culture of a common saprophytic and geoactive fungus, Aspergillus niger, in the presence of a solid source of calcium (Ca) and phosphorus (P). This observation, and our other data obtained, strongly suggests that fungal metabolism can provide a mechanism that can result in fossil biomineralization, and we hypothesize, therefore, that this may have contributed to the formation of well-preserved fossils (Lagerstätten) in the geological record. The characteristic polycrystalline nanofibers may also have served as a potential biosignature for fungal life in early Earth and extraterrestrial environments.

Ancient Basidiomycota in an extinct conifer-like tree, Xenoxylon utahense, and a brief survey of fungi in the Upper Jurassic Morrison Formation, USA

AbstractAlthough the well-known Upper Jurassic Morrison Formation has yielded abundant fossil plants for nearly a century, relatively little is known about fossil fungi and their ecological relationships to the Morrison flora. The first mention of fungal decay in fossil wood was briefly made over three decades ago, and since then, a few more reports of fungal decay associated with Morrison plants have been published. However, up to now, detailed data on the fossil fungi themselves have not been given from the Morrison Formation. Here we describe in detail well-preserved fossil mycelia in a silicified log of Xenoxylon utahense Xie et Gee, 2021 from the Upper Jurassic Morrison Formation at Miners Draw, Blue Mountain, near Vernal in northeastern Utah, USA. The fungal hyphae are variable in form, ranging from straight to slightly curved to highly coiled to tubular; they measure ~1.53 μm in diameter and possess clamp connections, septa, and occasional bifurcations. The occurrence of clamp connections typical of living Basidiomycota indicates a taxonomic affinity to this division of fungi. On the basis of the patterns of wood decay in the Xenoxylon log, the fossil fungi are interpreted here as pertaining to saprotrophic, white-rot wood fungi. These fossil mycelia represent a new record of ancient Basidiomycota from the Upper Jurassic Morrison Formation and provide further evidence for plant–fungus interactions in Jurassic terrestrial ecosystems.

The fungal ecology of the Brassington Formation (Middle Miocene) of Derbyshire, United Kingdom, and a new method for palaeoclimate reconstruction

Fossil fungi from periods warmer than modern climates provide unique insights into the future impacts of anthropogenic climate change. Here we report the fossil fungal assemblage from the late Middle Miocene Kenslow Member of central England, associated with climatic conditions warmer than the present-day. The identification of 110 morphotypes, which primarily relate to moist environments and the presence of wood, have been used to develop a new nearest living relative palaeoclimate reconstruction. The fungal assemblage indicates a Köppen–Geiger climate class, represented by temperate conditions, no dry season, and warm summers. This new fungal-based palaeoclimate reconstruction technique holds exciting potential to explore critically important but poorly understood palaeoenvironments, and the resulting qualitative inferences align well with previously published palaeobotanical quantitative estimates of palaeoclimate. These findings show that diverse fungal assemblages can successfully be used to reconstruct past climates for the first time.

(142–148) Proposals to improve the definition, utility, and curation of (type) specimens of fossil algae, fungi, and plants

(142–148) Proposals to improve the definition, utility, and curation of (type) specimens of fossil algae, fungi, and plants

Host Shift Speciation of the Ectomycorrhizal Genus Suillus (Suillineae, Boletales) and Biogeographic Comparison With Its Host Pinaceae.

Suillus is a genus of ectomycorrhizal fungi associated almost exclusively with Pinaceae. Lack of sample collections in East Asia and unresolved basal phylogenetic relationships of the genus are the major obstacles for better understanding the Suillus evolution. A resolved phylogeny of Suillus representing global diversity was achieved by sequencing multiple nuclear ribosomal and protein coding genes and extensive samples collected in East Asia. Fungal fossils are extremely rare, and the Eocene ectomycorrhizal symbiosis (ECM) fossil of Pinus root has been widely used for calibration. This study explored an alternative calibration scenario of the ECM fossil for controversy. Ancestral host associations of Suillus were estimated by maximum likelihood and Bayesian Markov chain Monte Carlo (MCMC) analyses, inferred from current host information from root tips and field observation. Host shift speciation explains the diversification of Suillus major clades. The three basal subgenera of Suillus were inferred to be associated with Larix, and diverged in early Eocene or Upper Cretaceous. In the early Oligocene or Paleocene, subgenus Suillus diverged and switched host to Pinus subgenus Strobus, and then switched to subgenus Pinus four times. Suillus subgenus Douglasii switched host from Larix to Pseudotsuga in Oligocene or Eocene. Increased species diversity occurred in subgenus Suillus after it switched host to Pinus but no associated speciation rate shifts were detected. Ancestral biogeographic distributions of Suillus and Pinaceae were estimated under the Dispersal Extinction Cladogenesis (DEC) model. Ancestral distribution patterns of Suillus and Pinaceae are related but generally discordant. Dispersals between Eurasia and North America explain the prevalence of disjunct Suillus taxa.

Identification of Chitin in Pliocene Fungi Using Py-GC × GC-TOFMS: Potential Implications for the Study of the Evolution of the Fungal Clade in Deep Time.

Molecular dating estimates the origin of the fungal clade to the Pre-Cambrian. Yet, the oldest unambiguous fungal fossils date to the Ordovician and show remarkable diversity and organizational development. Recent studies have suggested that the dates for the emergence of fungi in the fossil record may be pushed back to the Proterozoic. However, the nonspecificity of the methods used in those studies necessitates the employment of a wider variety of analytical techniques that can independently verify the presence of chitin, a crucial prerequisite in the assignment of fungal affinity, particularly of putative fossils from the Pre-Cambrian. In this paper, we propose Py-GC × GC-TOFMS as an example of one such technique. We analyze fungal fossils from the Pliocene. We find that a suite of N-bearing compounds are present in the pyrolysis products of these fossils, from which we suggest that 3-acetamidopyrones and their methylated homologues can serve as specific pyrolytic markers for chitin. We discuss both how this technique can potentially be used to differentiate between biopolymers, including those similar to chitin such as peptidoglycan, and the potential implications of identifying such markers in fossils from deep time. We conclude that Py-GC × GC-TOFMS is a promising technique that can potentially be used alongside, or independent of, staining methods to detect the presence of chitin in fossils.

Introducing palaeolithobiology

ABSTRACT A growing literature of deep but also surficial fossilized remains of lithobiological life, often associated with igneous rocks, necessitates the unfolding of a sub-discipline within paleobiology. Here, we introduce the term paleolithobiology as the new auxiliary sub-discipline under which fossilized lithobiology should be handled. We present key criteria that distinguish the paleolithobiological archive from the traditional one and discuss sample strategies as well as scientific perspectives. A majority of paleolithobiological material consists of deep biosphere fossils, and in order to highlight the relevance of these, we present new data on fungal fossils from the Lockne impact crater. Fungal fossils in the Lockne drill cores have been described previously but here we provide new insights into the presence of reproductive structures that indicate the fungi to be indigenous. We also show that these fungi frequently dissolve and penetrate secondary calcite, delineating the role lithobionts plays in geobiological cycles. We hope that the formalization of the sub-discipline paleolithobiology will not only highlight an overlooked area of paleobiology as well as simplify future studies of endo- and epilithic fossil material, but also improve our understanding of the history of the deep biosphere.

Celebrating Women in Paleobotany: A Tribute to Edith L. Taylor—Introduction and Dedication

Celebrating Women in Paleobotany: A Tribute to Edith L. Taylor—Introduction and Dedication

<strong><em>Pandea</em>, a new replacement name for <em>Udaria</em> D.K. Singh, S. Majumdar & D. Singh (Marchantiophyta, Lophocoleaceae)</strong>

The generic name Udaria Gupta (1996:103) was established by Gupta (1996) with two fossil fungi species Udaria singhii Gupta (1996:103) and U. saxenae Gupta (1996:104) from Lower Tertiary sediments of Himachal Pradesh, India. Later on, Singh et al. (2018) described the new genus of liverworts, Udaria Singh, Majumdar & Singh (2018: 1537) with the single species Udaria lamellicaulis Singh, Majumdar & Singh (2018: 1537) to the family Lophocoleaceae Vanden Berghen (1956: 208) from Arunachal Pradesh and Sikkim in Eastern Himalaya, India. After a thorough scrutiny of literature and type specimens, it was found that the liverwort genus name, Udaria Singh, Majumdar & Singh (2018: 1537) is illegitimate as it is a later homonym of the fossil fungi genus Udaria Gupta (1996:103). Therefore, a new replacement name Pandea U. B. Deshmukh is proposed with a new combination for the type species here in accordance with article 53.1 of Shenzhen Code (Turland et al. 2018).

The Pliocene Ixtacamaxtitlán low sulfidation epithermal deposit (Puebla, Mexico): A case of fossil fungi consortia in a steam-heated environment

The Ixtacamaxtitlán area in northern Puebla (central Mexico) contains middle Miocene Cu-Mo-Au porphyry/skarn and Pliocene low-sulfidation Au-Ag epithermal deposits that are geologically associated with the evolution of the Trans-Mexican Volcanic Belt (TMVB). In this paper, a new 40Ar/39Ar age (2.87 ± 0.41 Ma) is provided for rhombohedral alunite from a kaolinite + alunite ± opal ± cristobalite ± smectite advanced argillic alteration assemblage. This age contributes to the definition of a metallogenic province that is confined to the TMVB, a relevant feature for regional exploration. A ~12 My gap is established between the formation of the Cu-Mo-Au porphyry/skarn and low-sulfidation Au-Ag epithermal deposits, which rules out the possibility that their overlapping was the result of telescoping. Advanced argillic alteration is conspicuous throughout the mineralized area. This alteration assemblage consists of a widespread kaolinite-rich blanket that underlies silica sinters, polymictic hydrothermal breccias, and an alunite-rich spongy layer that consists of vertical tubular structures that are interpreted as the result of gas venting in a subaerial environment. The above indicate a shallow hypogene origin for the advanced argillic alteration assemblage—that is, formation by the partial condensation within a phreatic paleoaquifer of acidic vapors that were boiled-off along fractures that host epithermal veins at depth. The formation of the spongy alunite layer and silica sinters is interpreted to have been synchronous. Within the alunite-rich spongy layer, tubular structures hosted microbial consortia dominated by fungi and possible prokaryote (Bacteria or Archaea) biofilms. Such consortia were developed on previously formed alunite and kaolinite and were preserved due to their replacement by opal, kaolinite, or alunite. This means that the proliferation of fungi and prokaryotes occurred during a lull in acidic gas venting during which other organisms (i.e., algae) might have also prospered. Periodic acidic gas venting is compatible with a multi-stage hydrothermal system with several boiling episodes, a feature typical of active geothermal systems and of low-sulfidation epithermal deposits. The microstructures, typical for fungi, are mycelia, hyphae with septa, anastomoses between branches, and cord-like groupings of hyphae. Possible evidence for skeletal remains of prokaryote biofilms is constituted by cobweb-like microstructures composed of <1 µm thick interwoven filaments in close association with hyphae (about 2.5 µm thick). Bioweathering of previously precipitated minerals is shown by penetrative biobrecciation due to extensive dissolution of kaolinite by mycelia and by dissolution grooves from hyphae on alunite surfaces. Such bioweathering was possibly predated by inorganically driven partial dissolution of alunite, which suggests a lull in acidic gas venting that allowed living organisms to thrive. This interpretation is sustained by the occurrence of geometrical dissolution pits in alunite covered by hyphae. Fungal bioweathering is particularly aggressive on kaolinite due to its relatively poor nutrient potential. Such delicate microstructures are not commonly preserved in the geological record. In addition, numerous chalcopyrite microcrystals or microaggregates are found within the alunite layer, which could be related to sulfate reduction due to bacterial activity from the sulfate previously released by fungal bioweathering of alunite. Hydrogeochemical modeling constrains pH to between ~3.2 and ~3.6 and temperature to between 53 and 75 °C during the stage in which fungi and other organisms thrived. These waters were cooler and more alkaline than in earlier and later stages, which were characterized dominantly by steam-heated waters. The most likely process to account for this interlude would be mixing with meteoric water or with upwelling mature water that did not undergo boiling.

Diverse epiphyllous fungi on Cunninghamia leaves from the Oligocene of South China and their paleoecological and paleoclimatic implications

AbstractThe unique co‐occurrence of thyriothecia belonging to three fossil genera of epiphyllous fungi, Stomiopeltites Alvin & Muir (Micropeltidaceae), Callimothallus Dilcher, and Trichothyrites Rosendahl (Microthyriaceae), are reported on the leaves of the same host plant, Cunninghamia shangcunica Kodrul, Gordenko & Sokolova from the Oligocene Shangcun Formation of the Maoming Basin, South China. In China, Stomiopeltites is identified for the first time, Callimothallus is known from the Oligocene and Miocene of Guangxi and Zhejiang provinces, and Trichothyrites previously has been found only in the Eocene palynological assemblages of the Maoming Basin. The presence of abundant and diverse epiphyllous micromycetes, together with the taxonomic composition of the Shangcun megaflora and pollen assemblage, as well as quantitative climatic estimates obtained using Climate Leaf Analysis Multivariate Program, confirm the existence of a warm and humid climate in this region during the late early Oligocene. The geographic and stratigraphic distributions, comparisons with extant analogues, as well as ecological and paleoclimatic implications of the fossil fungi are discussed.

Molecular identification of fungi microfossils in a Neoproterozoic shale rock.

Precambrian fossils of fungi are sparse, and the knowledge of their early evolution and the role they played in the colonization of land surface are limited. Here, we report the discovery of fungi fossils in a 810 to 715 million year old dolomitic shale from the Mbuji-Mayi Supergroup, Democratic Republic of Congo. Syngenetically preserved in a transitional, subaerially exposed paleoenvironment, these carbonaceous filaments of ~5 μm in width exhibit low-frequency septation (pseudosepta) and high-angle branching that can form dense interconnected mycelium-like structures. Using an array of microscopic (SEM, TEM, and confocal laser scanning fluorescence microscopy) and spectroscopic techniques (Raman, FTIR, and XANES), we demonstrated the presence of vestigial chitin in these fossil filaments and document the eukaryotic nature of their precursor. Based on those combined evidences, these fossil filaments and mycelium-like structures are identified as remnants of fungal networks and represent the oldest, molecularly identified remains of Fungi.