Diversity of Ectomycorrhiza on Planted Seedlings in Variable Retention Forestry Sites: Results of Mushroom Survey

Diversity of ectomycorrhiza on planted seedlings in variable retention forestry sites: Results of mushroom survey.

Report January 2003

Renata Outerbridge, and J.A. Trofymow Pacific Forestry Centre, Canadian Forest Service Victoria, B.C.

Glen Dunsworth and Bill Beese Weyerhaeuser Coastal BC Group, Nanaimo, B.C. Forest Project – Silviculture and adaptive management program and Fred Bunnell Dept. Forest Science University of British Columbia

Contact: Dr. J.A. Trofymow, Phone 250 363 0677, email [email protected] Canadian Forest Service, Pacific Forestry Centre, 506 Burnside Rd. W. Victoria, BC. V8Z 1M5 2

Introduction

Unlike vascular plants, macrofungi of British Columbia are little known and documented in literature. In 1976, Bandoni and Szczawinski published the second edition of their concise pictorial guidebook to BC mushrooms. Gamiet and Berch, (1992) provide a species list from an old growth forest in Vancouver area. Countess (2001) investigated the impact of clear-cutting and forest age on macrofungi in old-growth and post-harvest chronosequences of Douglas-fir dominated in Shawnigan Lake District. Goodman (1995) and Goodman and Trofymow (1998) had previously used the same Douglas-fir stands as Countess (2001) to study distribution of ectomycorrhizal morphotypes. Outerbridge (2002) analyzed diversity and abundance of ectomycorrhizal and saprobic mushrooms in 35 year old monocultures of four conifer species on south Vancouver Island. Redhead (1997) estimated that there may be as many as 2500 species in the group of fleshy mushrooms (i.e. agarics, boleti, chanterelles) in the province and lists all the recorded reports. These amount to circa 500 species, a fraction of the potentially occurring diversity.

The purpose of this study was to survey sporocarps of macrofungi in Variable Retention (VR) forestry sites on southeastern Vancouver Island. The survey was proposed in conjunction with the study on ectomycorrhizal abundance and diversity in operationally planted seedlings in VR sites (Outerbridge et al., 2001), as part of Weyerhaeuser Coastal BC Project – Silviculture Monitoring (Beese, 2001). Generally, information on the presence of sporocarps is desirable whenever studying ectomycorrhizae in a forest ecosystem. Linking a sporocarp (the sexual structure) to its underground vegetative growth, the ectomycorrhizae, is an essential step in fungal taxonomy (Goodman et al, 1996). Mushrooms spread through an area via underground hyphal growth, or by animal aided dispersal, but most efficiently, via spore dissemination. The intent in initiating this survey was to facilitate the process of ectomycorrhizal species identification in our research on ectomycorrhizal diversity on the out-planted Douglas-fir seedlings, working on the assumption that the forest patch is a reservoir of the fungi found on their roots. 3

Some species (‘morphotypes’) from the pilot study are still unidentified and more are expected to emerge from the follow-up experimental study. By including saprophytic macrofungi in the survey we also hoped to give the data set merits of its own, independent of the ectomycorrhizae project. Provided the survey is repeated in subsequent year(s), the fungi can be looked at as another indicator group, for testing the effects of variable retention on biodiversity on these sites.

Methodology:

Seedlings of Douglas fir were planted in fall of 2000 at 6 sites (3 in old growth forest and 3 in second growth forest), along 3 transects at each site, at 4 stations on each transect 5m, 15m, 25m, and 45m from the edge of a forest patch (Outerbridge et al. 2001 and Outerbridge and Trofymow 2003, for site information). Two trips were made to each site during the fall period (October 7 – 9; November 18 – 25, 2002) to carry out a mushroom inventory in the forest patch at the start of each transect. Macrofungal species were recorded using the absence/presence method by surveying a rectangular areas 5m in width starting at patch edge and extending 10m into the patch. Mushrooms were not surveyed on the transect. All fleshy fungi (ectomycorrhizal and saprobic), minimum 1cm large, were documented. We examined the forest floor as well as woody debris, tree stumps, decaying logs etc., making notes on the nature of substrate for each mushroom recorded. Unless spotted at the soil surface, subterraneous sporocarps (such as true and false truffles) were excluded. For some species, identification was possible in the field, others were collected, identified in the lab and preserved as voucher specimens at Pacific Forestry Centre. The inventory was carried out following the guidelines in Readhead and Berch (1996). Taxonomic literature used included: Breitenbach and Krunzlin vol. 1-4 (1984-195), Arora (1986), Smith (1947), Smith et al. (1979), Smith et al. (1981), and Phillips (1991). 4

Results

The October field trip yielded a total of 25 species of macrofungi, with 13 of those in the genus Mycena (Table 1). Only one mycorrhizal species was found, Hydnum fuscoindicum, in B1 site. The remaining mushrooms were small sized litter and wood decomposers. Fomitopsis pinicola is listed as a species of pharmaceutical value (Dr. S.Berch, personal communication), and Phaeollus schweinitzii is a wood rotting pathogen. An edible jelly-like species, Tremella mesenterica, or Witch’s Butter, was recorded in one plot in R23, but a fair amount of this fungus was also spotted outsite the plots in B1 site.

A total of 39 species of macrofungi were recorded during the November field trip to the same plots (Table 2). Again, about half the species belong to the genus Mycena. The number of ectomycorrhizal mushrooms was still low, with only 5 species found: Hygrophorus sp., Inocybe napipes, Laccaria sp., Laccaria laccata, and Tricholoma sp. The remaining species were litter decomposers and woody substrate dwellers. Among these the most common ones were Marasmius salalis, with its characteristic strong garlic odor, Mycena alcalina (bleachy smell), and a small, rather non-descript, Mycena murina, quite common in the Pacific Northwest forests. Pseudohydnum gelatinosum, or Toothed Jelly Fungus, is edible, but too infrequent to be of commercial value.

The cumulative occurrence of all macrofungi at each site along with guild/substrate group is presented in Table 3. Overall, 16 species repeated fruiting, with 12 of those found again in the same locations (two of these, however, Fomitopsis pinicola and Nidula candida form tough long lasting fruiting bodies, and therefore most likely persisted from the first to the second survey). Most common species found in second growth forest patches were: Mycena alcalina, Mycena murina group, Nidula candida, and Strobilurus trulisatus, while the Old Growth was characterized by two more frequent species: solitary Marasmius salalis and troop forming Mycena strobilinoides. 5

Pooled data from both surveys shows a total number of 151 occurrences represented by 48 species of macrofungi (Table 4). The highest values in both measurements were in DAM 1 and the lowest in C1500. Overall, second growth forest patches yielded more mushrooms than the old growth patches.

Six guilds of macrofungi could be distinguished based on their growth substrate. They were in order of their predominance: litter decomposers, wood decaying fungi, general decomposers, ectomycorrhizae, fungi associated with bryophytes (basidiolichens), and one species which specializes in growing on Douglas-fir cones (Table5). Ectomycorrhizal fungi accounted for 12.5% of the overall species richness.

Discussion

It is hard to predict or to give an accurate account of the diversity of macrofungus species on a site. Sporocarps of these organisms can be studied using similar methods as those used in botany. However, unlike plants, most fungal sporocarps do not persist in the environment for very long. It is necessary therefore to carry out repeated surveys of the same site in order to obtain the full picture of its mycobiota. This is especially true during years with unusual weather patterns. The fall of 2002 in southwestern Vancouver Island was very dry, more so than in several previous years. It should be stressed, therefore, that the results obtained here most likely underestimate the actual macrofungal diversity present in the areas studied. Countess (2001) reported 384 species of macrofungi observed from 1995 to 1997 in Douglas fir dominated CWHxm forests. Also on south Vancouver Island, Outerbridge (2002) found 277 species in a two-year survey of selected conifer monoculture forests, with 142 of those observed in Douglas-fir stands.

Two factors are most important in regulating fungal fruiting: temperature and soil moisture (Carrol and Wicklow, 1992). Usually, the best scenario for large crop of mushrooms involves below-ground build up of mycelium during steady, warm and moist 6 early fall followed by a rapid drop in temperature and abundant rain later in the season. The fall of 2002 did not meet these conditions. There were many cool, dry and windy days in October, which dried out the forest floor and the situation was exacerbated by the lack of rain and no significant drop in temperature in November.

Therefore it is hard to discuss any patterns in diversity of mushrooms in the VR forest patches or to try and apply the meager data to our research on mycorrhizae at these sites. It would be interesting to know whether the fungi listed here as “most common”in second growth stands are indeed the dominant species, or whether they are less common in old growth stands. Similarly, we would like to know if Mycena strobilinoides is found exclusively in old growth stands. It is likely that this species prefers higher elevation (all old growth plots in this study are at higher elevation than the second growth ones), but the seral stage affinity is also possible.

In southern British Columbia forests, ectomycorrhizal fungi typically comprise a lower percentage of macrofungus flora than the other quilds combined (Berch and Gamiet, 1992; Countess, 2001; Outerbridge, 2002), but can be the largest group if the guilds are looked at separately (Countess, 2001). In either case, the percentage of ectomycorrhizal species is seldom below 30%. In this survey we found an unusually low number of ectomycorrhizal species, i.e. only 6 or 12.5% of the whole mycobiota, undoubtedly caused by insufficient moisture level in the soil. The generally smaller, less fleshy macrofungi of the litter or wood decomposer groups, such as those in the genus Mycena, can extract the small amounts of water bound in forest duff or decaying wood and form fruiting bodies. Thus, they are the first ones to appear after a short rainfall, though do not persist for more than a few days. Sporocarps of a few tough species, such as conks (e.g. Fomitopsis), or Bird’s Nests (e.g. Nidula), can usually be found all year round, regardless of precipitation level.

Countess found a higher proportion of macrofungi in ectomycorrhizal group (123 species) than in other guilds (90 litter decomposers, 28 general saprobes, 51 wood decomposers, 1 basidiolichen, and 1 parasitic species) in the month of October 1995, 7

1996, and 1997. He observed wide variation in species richness between the three years, with ectomycorrhizal fungi accounting for 47, 43, and 102 species in the first, second and third year respectively. Goodman (1995) described 69 types of ectomycorrhizae comparing old-growth and mature stands of Douglas-fir. He also stated that the similarity of the ectomycorrhizal species diversity and composition between the two forest age groups was probably due to their proximity and the similarity of their vegetation and soil. In our study equal number of ectomycorrhizal macrofungi (3 species) were found in second growth and in old growth stands, and clearly, it is too early to draw any conclusions.

In retrospect, with regards to ectomycorrhizal mushrooms, it would be more helpful if soil samples are collected along with the sporocarps, so as to be able to compare the mycelium from the roots of trees in the forest patch with the various morphotypes observed in the ectomycorrhizae survey. We also suggest that hypogeous mushrooms be included in future surveys. These would include such genera as Rhizopogon, or Truncocolumella. They form ectomycorrhizae with Douglas-fir and were found on the roots of the pilot study seedlings, Rhizopogon being the most abundant of all the morphotypes. In an Oregon study, Smith et al. (2002) have found that as much as 18.25% of ectomycorrhizal symbionts of young, rotation-age, and old-growth stands of Douglas- fir were from the hypogeous group. More research is needed on these sites to obtain a better picture of mushroom diversity. 8

Table 1. Macrofungus inventory in VR forest sites, October 7-9, 2002.

Stand Age second growth old growth Total B1 B1 B1 TL TL TL R23 R23 R23 C1500 C1500 C1500 H1220 H1220 H1220 Site Name DAM 1 DAM 1 DAM 1 Transect 123123123123123123 Fomitopsis pinicola 1 1 Galerina vittaeformis 1 1 Guepiniopsis alpina 1 1 Hydnum fuscoindicum 1 1 Marasmiellus candidus 1 1 Mycena alcalina 1 1 Mycena capillaris group1 1 Mycena epipterygia 1 1 Mycena filopes-like 1 1 Mycena leucogala-like 1 1 Mycena lilacifolia 1 11 3 Mycena metata group 1 1 Mycena murina group 11 2 Mycena pura 1 1 Mycena rorida 111 1 1 1 6 Mycena scabripes 11 Mycena sp. 1 1 Mycena strobilinoides 111 3 Nidula candida 11 11 1 5 Omphalina pyxidata-like 1 1 Phaeollus schweinitzii 1 1 Psathyrella longistriata 1 1 Strobilurus trulisatus 111 3 Tremella mesenterica 1 1 Xeromphalina fulvipes 1 1 1 3 Number of occurences = number of species 22912113334241013143 9

Table 2. Macrofungus inventory in VR forest sites, November 18 – 25, 2002. Species in bold re-occurred.

Stand Age second growth old growth Total B1 B1 B1 TL TL TL R23 R23 R23 C1500 C1500 C1500 H1220 H1220 H1220 Site Name DAM 1 DAM 1 DAM 1 Transect 123123123123123123 Collybia dryophila 1 1 Fomitopsis pinicola 1 1 Galerina heterocystis group 1 1 1 3 Galerina vittaeformis 1 1 Guepiniopsis alpina 1 1 111 5 Hygrophorus sp. 1 1 Inocybe napipes 1 1 Laccaria sp. 1 1 Laccaria laccata 1 1 Marasmius salalis 11 1 1 11111 1111 Micromphale perforans 1 1 Mycena alcalina 1 111 111 1 1 9 Mycena amabilissima 1 1 Mycena clavicularis 1 1 Mycena delicatella 11 2 Mycena epipterygia 1 11 3 Mycena filopes-like 111 11 5 Mycena fusco- occula 1 1 Mycena galopus 1 1 Mycena haematopus 11 2 Mycena lilacifolia 1 1 1 3 Mycena metata group 111 3 Mycena murina group 11111 111 1 110 Mycena pura 1 1 Mycena rorida 1 1114 Mycena rosella 11 2 Mycena rubromarginata 1 1 2 Mycena sp. 1 11 3 Mycena strobilinoides 11111 5 Nidula candida 11 11 4 Omphalina ericetotum 1 1 Panellus stipticus 1 1 Panellus longinquus 1 1 Psathyrella longistriata 1 1 Pseudohydnum gelatinosum 1 1 Strobilurus trulisatus 1 1111 1 6 Tricholoma sp. 1 1 Xeromphalina fulvipes 1 1111 1 6 Xylaria hypoxylon 1 1 Number of observations = number of species 14610895538343643575108 10

Table 3. List of all macrofungi found fruiting at VR sites in Oct7-Nov25, 2002. second old growth growth Total Site Name Guild / Substrate DAM 1 TL B1 R23 C1500 H1220 occurrences Collybia dryophila litter 0 0 1 0 0 0 1 Fomitopsis pinicola wood 2 0 0 0 0 0 2 Galerina heterocystis group wood 2 1 0 0 0 0 3 Galerina vittaeformis basidiolichen 2 0 0 0 0 0 2 Guepiniopsis alpina wood 0 0 1 2 3 0 6 Hydnum fuscoindicum ectomycorrhizal 0 0 1 0 0 0 1 Hygrophorus sp. ectomycorrhizal 0 0 1 0 0 0 1 Inocybe napipes ectomycorrhizal 0 0 0 0 0 1 1 Laccaria sp. ectomycorrhizal 0 0 0 1 0 0 1 Laccaria laccata ectomycorrhizal 0 0 1 0 0 0 1 Marasmiellus candidus wood 1 0 0 0 0 0 1 Marasmius salalis litter 2 1 1 2 3 2 11 Micromphale perforans litter 0 0 1 0 0 0 1 Mycena alcalina litter/wood 3 2 3 0 0 2 10 Mycena amabilissima litter 0 1 0 0 0 0 1 Mycena capillaris group litter 1 0 0 0 0 0 1 Mycena clavicularis litter 0 0 0 1 0 0 1 Mycena delicatella litter/wood 2 0 0 0 0 0 2 Mycena epipterygia litter 1 0 0 1 2 0 4 Mycena filopes-like litter/wood 1 2 0 0 0 3 6 Mycena fusco- occula litter 1 0 0 0 0 0 1 Mycena galopus litter/wood 0 1 0 0 0 0 1 Mycena haematopus wood 2 0 0 0 0 0 2 Mycena leucogala-like wood 0 0 0 1 0 0 1 Mycena lilacifolia wood 0 0 2 1 2 1 6 Mycena metata group litter 1 0 0 0 1 2 4 Mycena murina group litter 3 2 5 0 1 1 12 Mycena pura litter/wood 1 1 0 0 0 0 2 Mycena rorida litter 0 0 4 1 2 3 10 Mycena rosella litter 0 0 2 0 0 0 2 Mycena rubromarginata litter 0 1 0 0 0 0 1 Mycena scabripes litter 0 0 0 0 0 1 1 Mycena sp.1 litter 1 0 0 0 0 0 1 Mycena sp.2 litter 1 2 0 0 0 0 3 Mycena strobilinoides litter 0 0 0 6 1 2 9 Nidula candida wood 4 4 0 0 1 0 9 Omphalina pyxidata-like basidiolichen 0 0 0 0 1 0 1 Omphalina ericetorum basidiolichen 1 0 0 0 0 0 1 Panellus stipticus wood 1 0 0 0 0 0 1 Panellus longinquus wood 1 0 0 0 0 0 1 Phaeollus schweinitzii wood 0 0 0 0 0 1 1 Psathyrella longistriata litter 2 0 0 0 0 0 2 Pseudohydnum gelatinosum wood 1 0 0 0 0 0 1 Strobilurus trulisatus Fd cones3 50001 9 Tremella mesenterica wood 0 0 0 1 0 0 1 Tricholoma sp. ectomycorrhizal 0 0 0 1 0 0 1 Xeromphalina fulvipes wood 2 3 0 1 1 2 9 Xylaria hypoxylon wood 1 0 0 0 0 0 1 Total occurrences 43 26 23 19 18 22 151 11

Table 4. Total and per site macrofungus diversity.

Site Total number of Total number Stand Age Name occurrences of species

Second Growth DAM 1 43 26

Second Growth TL 26 13

Second Growth B1 23 12

Old Growth R23 19 12

Old Growth C1500 18 11

Old Growth H1220 22 13

Total S.G. 92 37 Total O.G. 59 22

All Sites 151 48

Table 5. Ecological guilds of macrofungi in VR forest sites.

Number of Number of Species in Species in Old Total Number of Guild / Substrate Second Growth Growth Species % of Total Litter / Humus 15 8 18 37.5 Wood 12 7 15 31.25 Ectomycorrhizal 33 6 12.5 Litter and Wood 52 5 10.42 Basidiolichen 21 36.25 Cones 11 12.1

Acknowledgements 12

This study was funded by an NSERC grant to University of British Columbia (Dr.F.Bunnell) as part of a larger project which was developed through the Forest Project initiative of Weyerhaeuser’s BC Coastal Group. Additional funding for the field survey came from the Biodiversity Network of the CFS. We gratefully acknowledge the field assistance of Bob Ferris (CFS).

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