Macrofungi of Two Peoples Bay Nature Reserve, Western Australia

Introduction

Fungi are found everywhere in the environment, where they play a crucial role in maintaining healthy ecosystems. Fungi have three basic life modes: they are saprotrophs, symbionts, or parasites. The saprotrophic fungi (rotters) break down organic matter such as wood and leaf litter. Symbionts, such as mycorrhizal fungi, form a mutually beneficial (mycorrhizal) partnership with more than 90% of plant families, a role which involves the exchange of minerals, water and carbohydrates (Brundrett et al. 1996; Tommerup and Bougher 1999). Lichens form mutually beneficial partnerships with green algae and cyanobacteria. Parasites derive their nutrients from other living organisms; in the macrofungi (those large enough to see without a visual aid) parasites form the smallest of the three groups. Some fungi are facultative parasites and can survive without a host, usually as a saprotroph (Pascoe and Shipton 1996).

What we see and use for identification is the dispersal or spore bearing structure of the fungus, the sporocarps. However, the main vegetative portion of a fungus (mycelium) is composed of microscopic filaments (hyphae) that ramify through the substrate, leaking enzymes that break down organics and minerals, then absorbing nutrients and water back into the mycelium. Most of the macrofungi sporocarps we see are the above ground, or epigeous sporocarps, fruiting on soil, leaf litter or woody debris, on stems or trunks, or in the canopy of plants, and more rarely on insects. Buried at various depths (or at times emergent) are the hypogeous, truffle-like or sequestrate fungi, most of which are also mycorrhizal. Such fungi generally cannot release their spores and therefore require the assistance of vertebrates or insects for spore dispersal (Fig. 1).

Fig. 1.

Examples of different sporocarp forms. (a) Artomyces austropiperatus (coral); (b) Lactifluus flocktoniae (stalked lamellate); (c) Cortinarius luteirufescens (truffle-like or sequestrate); (d) Podoserpula pusio, the pagoda fungus; (e) Cordyceps sp. (parasitic); (f) Antrodiella citrea (bracket); (g) Poronia erici or dung buttons; (h) Geastrum sp. (puffball-earthstar) (photographs K. Syme).

Australia is estimated to have from 180,000 to 250,000 species of fungi, a number that includes 8000 to 12,000 macrofungi (Pascoe 1991; May and Wood 1997; May 2017). It is estimated that as few as 15–20 percent of Australian macrofungi are named (May and Wood 1997; Bougher and Lebel 2001; Newell et al. 2008; May 2017), so any list of fungi will necessarily contain many unknown genera and a greater number of un-named species than named ones. Prior to 2003, Western Australia was one of the few states that had a relatively recent checklist of macrofungi, for which all records were vouchered (Hilton 1982, 1988). Since then comprehensive attempts have been made to update census/checklists for other states and territories, based upon literature records and digitisation of data for holdings in state herbaria. There is now a National Species List for fungi (https://fungi.biodiversity.org.au/nsl/services/search/taxonomy) and lichens (https://lichen.biodiversity.org.au/nsl/services/search/taxonomy), enabling consistency in name usage and taxonomic updates to be taken up nationally.

Some fungi produce sporocarps only sporadically. Therefore, a continuous sampling over many years is necessary to gain a thorough picture of the range of species of fungi inhabiting any given area or vegetation type, together with the provision of good quality collections for herbaria. With new molecular tools enabling sampling of soil from different habitats or animal faecal pellet samples, it has become possible to generate data that reveal the diversity of fungi, past and present at a site (Bastias et al. 2006; Hughes et al. 2009) or provide insight into the diet of small mammals (Bannister et al. 2021; Hopkins et al. 2021). However, these data are highly reliant on strong taxonomic research of fungal lineages to provide backbone data for comparison. The lack of expert Australian mycologists to interpret these data continues to be an impediment. For Australia, there are very few lists of the fungi inhabiting particular sites based on repeated sampling (T. May, pers. comm. 2004; Gates et al. 2005; Robinson and Tunsell 2007; Catcheside and Catcheside 2008; Robinson et al. 2008; Catcheside et al. 2009). This has improved in the past two decades due to the establishment of the citizen science based Fungimap project in 1995, an Australia-wide mapping effort of 100 target species, followed by the formation of several regional mycological societies in Queensland (Queensland Mycological Society), Victoria (Field Naturalists Club of Victoria-Fungi Group), New South Wales (South Coast Fungi Group and Sydney Fungal Group), and South Australia (Adelaide Fungal Studies Group). The advent of iNaturalist records in the last decade has also improved recording of observational data.

Bougher et al. (1994) and May (2002) noted that as many as 75% of species of southern Western Australian (WA) macrofungi were also present in eastern Australia, and fewer than 60 endemic WA species (10%) had been identified. Most species were represented by fewer than 4–8 collections, with some known only from restricted locations. Similar relative proportions of endemic macrofungi were found for Victoria (May and Avram 1997; May 2002), but were higher for South Australia (SA), where more than 30% were considered to be endemic to that state (Grugurinovic and Simpson 2001).

Because of its unique, near-pristine state and the perceived richness of its fungi, Two Peoples Bay Nature Reserve has been the focus of mycological interest since 1974, when Dr Graeme Smith (CSIRO) sent collections of fungi he discovered in the Reserve to Roger Hilton, lecturer in mycology at the Botany Department, University of Western Australia. The re-discovery of Gilbert’s potoroo (Potorous gilbertii) or Ngilgaitch (Noongar names are mainly from Knapp et al. 2024) on the Maardjitup Gurlin/Mt Gardner headland by Elizabeth Sinclair in 1994 (Sinclair et al. 1996) focused attention on the sequestrate hypogeal fungi, because of the potoroo’s specialised enlarged foregut in which pre-gastric fermentation takes place. This adaptation is believed to aid extraction of nutrients, thus enabling potoroos or rat kangaroos to thrive on a diet consisting almost solely of fungi (Christensen 1980; Claridge and Cork 1994; Claridge and May 1994; Tory et al. 1997; Vernes 2007, 2010). Soon after the re-discovery of Ngilgaitch/Gilbert’s potoroo portions of their faecal pellets were examined microscopically for evidence of fungal spores and more than 23 different taxa were identified (K. Syme, 1995 unpubl. data). In a later study of further samples, 90% of material in the pellets was found to be fungal in origin and contained about 25 spore morphotypes. Some of these matched spores from fungal specimens collected in potoroo habitat (Bougher 1998). Most spores were from basidiomycete truffle-like sporocarps and the remainder were from ascomycete or epigeous fungi.

Fungi have been recorded and/or collected at Two Peoples Bay Nature Reserve by Roger Hilton, Dr Graeme Smith (CSIRO), Dr Derek Reid (Royal Botanic Gardens Kew, UK), Dr Neale Bougher (CSIRO, Perth), Professor OK Miller Jr (Blacksburg, Virginia, USA), Katrina Syme, Dr Teresa Lebel (then at the Royal Botanic Gardens Victoria, Melbourne), Dr Elaine M. Davison, Dr Brandon Matheny (University of Tennessee, USA), and Alan Danks, with one to three collections by Linda Reinhold, Mattias Ryberg, Jennifer Tonkin, C & K Vankey, and G. & R. Byrne. The Reserve was also the focus of two 12-month mycological surveys conducted by K. Syme, with the assistance of volunteers in 1991–1992 covering most major vegetation areas and focusing on hypogeal sequestrate fungi in 1998–1999 following the re-discovery of the Ngilgaitch/Gilberts potoroo 4 years earlier. In 2020, 5 years after the 2015 wildfire, the Gilberts Potoroo Action Group employed K. Syme to conduct targeted surveys in vegetation types and habitats where potoroo activity was known. Some examples of different sites and vegetation types surveyed during these projects are shown in Figs 2 and 3. More images sites and vegetation types in the Reserve are presented in Hopkins et al. (2024).

Fig. 2.

Some examples of sites and vegetation types surveyed for macrofungi. (a) Melaleuca scrub near Tick Flat track 2020; (b) Robinsons Gully 2020; (c) Wave Sign Gully Track 2007; (d) Webster’s Gully beyond Little Beach with Maardjitup Gurlin/Mt Gardner right of centre 2007; (e) picnic area near research Quarters 2007; (f) bushfire regeneration on lower section of Firebreak Valley Track 2020 (photographs K. Syme).

Fig. 3.

More examples of sites and vegetation types surveyed for macrofungi. (a) Eucalyptus megacarpa forest near research Station January 2024; (b) mixed heath high on Firebreak Valley Track (photographs A. S. Hopper, B. K. Syme).

Materials and methods

Herbarium and Fungimap data

Atlas of living Australia (ALA) data for fungal records in the Reserve were downloaded (30 April 2024), and non-macrofungal records removed. Species lists from surveys and inventories provided in various reports (many unpublished) were cross-checked against the ALA data and duplicates removed. Collections identified only to order or family were deleted and names then checked against the National Species list for Fungi (https://fungi.biodiversity.org.au/nsl/services/search/taxonomy) and updated to current classification. Where collections have not been able to be assigned to known species, a provisional name was used, indicated by a short tag name such as ‘Amanita abruptly bulbed’. Each of these provisional taxa was considered to represent a different species in initial lists. However, interpretation of these provisional names was problematic once collection data were examined, thus they were included under ‘Genus spp.’ in the final summary table. For collections where we were certain a single unknown species was present, we list it as ‘Genus sp.’ in the summary table. Twenty-four sites distributed in 17 different vegetation types were determined for all collections based on collection details; acronyms were devised, then applied for each species and included in the final summary table (sites shown on Fig. 4). A final category of ‘?’, signifying an undeterminable location was included for 35 collections where no GPS data was available, or location was simply listed as ‘Two Peoples Bay Reserve’. We acknowledge that the vegetation types listed for some sites is an oversimplification of the rich complexity of habitats at the location. However, it was not possible to tease out enough details from the collection notes or GPS data to follow the breakdown provided in Hopkins et al. (2024).

Fig. 4.

Map of Two Peoples Bay Nature Reserve showing collecting sites of macrofungi in green boxes. These include Webster’s Gully (WGu); Heritage Trail (Htb); Goodga River Track (GRtk); forest near Reserve Office (RO); Sinker Reef Track (SRf); Juniperina Creek (JCk); Wilson’s Track (WT); Firebreak Gully Track (FGTk); Firebreak Gully Valley (FbV); Kaiupgidjukgidj/Moates Lagoon (MLtk); Rocky Point Track (RcPt); Tyiurrtmiirit^y/Lake Gardner (Mela); Little Beach (LitBe); Robinson Gully (RoGu); Hakea Gully (HaG); Wave Sign Gully (WSGu); Fitzpatrick’s Track (Fitz); Rydene Property (RyDe); Maardjitup Gurlin/Mt Gardner (MtGa); Western Boundary Track (WBTr); Tick Flat (TcFtr); Area 5, off track from V4S (Area5); Little Bay (LiBy); Bishops Gully (BiGu). See Tables 1 and 2 for list of species collected. Base map by A. J. M. Hopkins and G. T. Smith.

Surveys and collecting events

An inventory of the macrofungi was conducted in the Reserve between June and September 1991 and from March to May 1992, the optimal season for fungi in this region, by K. Syme (Syme 1992). Sampling was carried out for up to 3 days each week along 31 different tracks, walk trails, gullies, and in dunes and woodland areas. However, this was reduced to 24 sites in the final summary table as several tracks and trails were located very close to each other and ran through the same general location and vegetation type, and also only had a single collection or observation recorded (Table 1). Visits to Djimaalap/noisy scrub-bird (Atrichornis clamosus) breeding areas (such as the gullies) were limited. Syme also carried out random collection surveys from 1998 to 1999 sampling 2 days/week along tracks and trails each month (Syme 1999), and again in 2007–2008 (assisted by Dr J. Newell in 2008), as part of a survey of fungi in the South Coast Natural Resource Management Region (Syme 2008). One-off collections or 1–2 day forays were also conducted at other times over the past four decades by K. Syme and a range of researchers. All road, track, and trails were accessed on foot and on all occasions, as a precaution against the spread of soil-borne pathogens, strict hygiene measures were respected (Hart et al. 2024).

Table 1.List of sites (acronyms) visited during macrofungi surveys and main vegetation types at each site (locations are shown on Fig. 4).

Acronym	Full site name	Associated vegetation/habitat
WGu	Webster’s Gully	Woodland deep leaf litter: Eucalyptus megacarpa, Eucalyptus calophylla, Melaleuca baxteri
Htb	Heritage trail	Mixed low woodland, deep leaf litter: Eucalyptus cornuta, Eucalyptus megacarpa, Agonis flexuosa OR Agonis flexuosa thicket OR Allocasuarina fraseriana, Melaleuca sp. OR Banksia sp.
GRtk	Goodga River Track	Open forest: Corymbia calophylla, Eucalyptus marginata, Allocasuarina
RQ	Research Quarters	Open woodland and heath: sandy areas
SRf	Sinker Reef track	Coastal shrubland: Agonis flexuosa, Melaleuca thymoides, Spyridium sp., Acacia cochlearis, Leucopogon sp., Olearia axillaris and other shrubs
JCk	Juniperina Creek	open forest wet: Eucalyptus calophylla and Agonis flexuosa, Melaleuca sp.
WT	Wilson’s Track	Low woodland, deep leaf litter: Eucalyptus cornuta, Agonis flexuosa, Spyridium globulosum, Oxylobium sp., Bossiaea sp.
FGTk	Firebreak Gully Track	Woodland: Eucalyptus marginata, Corymbia calophylla, Allocasuarina fraseriana, Oxylobium lanceolatum OR Melaleuca spp. and Agonis spp.
FbV	Firebreak Gully Valley	Open woodland or heathland: Allocasuarina fraseriana, Eucalyptus megacarpa, Eucalyptus marginata, Corymbia calophylla, Agonis hypericifolia
MLTk	Kaiupgidjukgidj/Moates Lagoon Tk	Woodland or heath: Eucalyptus patens, Eucalyptus staeri, Eucalyptus marginata, Allocasuarina fraseriana, Agonis hypericifolia, Melalueca thymoides
RcPt	Rocky Point	Coastal heath: Agonis flexuosa, Melaleuca thymoides, Allocasuarina humilis, Spyridium, Acacia cochlearis
Mela	Gardner Creek/Gardner Lake/Tyiurrtmiirity	Woodland: Melaleuca baxterii and Agonis flexuosa, Taxandria parviceps OR Coastal heath shrubland with Agonis spp. and shrubs of Myrtaceae and Fabaceae OR Banksia attenuata woodland
LitBe	Little Beach	Heath shrubland: Allocasuarina and myrtaceous shrubs, Agonis flexuosa, Spiridium globulosum, Acacia and Melaleuca
RoGu	Robinsons Gully	Open forest: Eucalyptus marginata, Eucalyptus megacarpa & Agonis flexuosa, Allocasuarina, Hakea
HaG	Hakea Gully	Open forest: Corymbia calophylla, Eucalyptus marginata, Taxandria parviceps
WSGu	Wave Sign Gully	Low woodland: Eucalyptus conferruminata, Eucalyptus calophylla, E. marginata (coastal form), Oxylobium cuneata, Hakea cucullata, Allocasuarina fraseriana, Agonis marginata, Agonis hypericifolia
Fitz	Fitzpatrick’s Track	Low woodland: Eucalyptus cornuta, Agonis flexuosa, Spyridium globulosum
RyDe	Rydene Property	Woodland: Eucalyptus marginata, Corymbia calophylla, Allocasuarina, Xanthorrhoea
MtGa	Wes’ Hilltop site, Maardjitup Gurlin/Mt Gardner	Heath: Gastrolobium bilobium
WBTr	Western Boundary Track	Woodland: Eucalyptus marginata, Melaleuca sp., Allocasuarina humilis, A. fraseriana, Agonis flexuosa
TcFtr	Tick Flat	Open heath: Dryandra sessilis, Agonis flexuosa
Area5	Area 5, off track from V4S	Swamp: Agonis flexuosa and Eucalyptus megacarpa.
LiBy	Little Bay	Coastal heath
BiGu	Bishops Gully	Woodland: Eucalyptus angulosa and Allocasuarina fraseriana
?	Uncertain	Woodland or heath shrubland: Eucalyptus, Agonis, Allocasuarina, Gastrolobium

Most species of fungi were gathered only once and where possible, all growth stages were included in the collection; associated plant species were recorded in the field and if known, standard vegetation codes applied. Unique code numbers were assigned to each collection and detailed fieldnotes of sporocarps recorded. The fungi were then dried in a fan-forced dehydrator and stored in clip lock plastic bags. Where applicable, spore prints were collected, individually labelled and packaged, and when required to assist identification, spores, and other microscopic characters were examined at 1000× magnification using a compound microscope (Figs 5 and 6).

Fig. 5.

Some microscopic characters useful to differentiate genera and species. (a) Ridged spores of Austrogautieria sp.; (b) smooth spores with inflated utricle of Hysterangium sp.; (c) metuloid crystals on pleurocystidia of Inocybe spp. (20 um scale bar = to lower elongated cell swollen base (side to side)); (d) amyloid ornamentation of Russulaceae spores (scale bar (10 Um) on the left side); (e) amyloid hyphal trama of a hypogeal bolete; (f) pileocystidia in pellis of Lactifluus clarkeae (50 um scale bar = to filamentous layer width) (Photographs a–c K. Syme; d–f T. Lebel).

Fig. 6.

Drawings made in 2020 by K. Syme of spores found in faecal pellets from two Ngilgaitch/Gilbert’s potoroos (Potorus gilbertii).

The first stage of a dietary study of the critically endangered Ngilgaitch/Gilbert’s potoroo began in February 1998 and was completed a year later (Syme 1999). Survey plots of 20 × 40 m were established in each of five different vegetation types: Forest, Heath/Sedge, Sedge, Woodland/Heath, and Forest/Heath, and sampled once per month. The Forest site was in Firebreak Valley (locations named are shown in Fig. 4), and the only one located near foraging sites of the potoroos; the remaining sites were some distance down slope, west of Firebreak Valley Track. Further studies of macrofungi were made in sites frequently visited by potoroos, as evidenced by diggings. These sites were surveyed by Syme and volunteers in 2020, 5 years after the 2015 wildfire.

Results