Canadian Journal of Forest Research

Fungi decaying the wood of fallen beech (Nothofagus) trees in the South Island of New Zealand.

Journal: Canadian Journal of Forest Research

Manuscript ID cjfr-2018-0179.R1

Manuscript Type: Article

Date Submitted by the 29-Aug-2018 Author:

Complete List of Authors: Hood, Ian; Scion (New Zealand Forest Research Institute) McDougal, Rebecca; Scion Somchit, Chanatda; Scion Kimberley, Mark; Scion (New Zealand Forest Research Institute) Lewis, Aymee;Draft Scion Hood, Joy; 25 Simmonds Crescent

fungal biodiversity, decomposer fungi, decay of fallen Nothofagus stems, Keyword: New Zealand indigenous forest ecology, basisiomycete wood colonisers

Is the invited manuscript for consideration in a Special Not applicable (regular submission) Issue? :

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1 Fungi decaying the wood of fallen beech (Nothofagus) trees in the South Island of New Zealand.

2

3 Ian A. Hood, Rebecca L. McDougal, Chanatda Somchit, Mark O. Kimberley, Aymee S.R. Lewis and Joy

4 O.L. Hood

5 I.A. Hood, R.L. McDougal, C. Somchit, M.O. Kimberley, and A. Lewis. New Zealand Forest Research Institute (Scion),

6 Private Bag 3020, Rotorua 3046, New Zealand.

7 J.O.L. Hood. 25 Simmonds Crescent, Rotorua 3015, New Zealand.

8 Abstract

9 10 In order to extend present knowledge of communities of wood decay fungi in native forests, basidiomycetes

11 and ascomycetes were isolated from within 15 fallen stems in beech (Nothofagus, Nothofagaceae) forests in

12 the South Island of New Zealand. Fungal species were identified as precisely as possible using traditional

13 culturing and molecular approaches. The internal distribution of species within stems was determined.

14 Common fungi that occupied significant portionsDraft of stems were Ganoderma applanatum sensu Wakefield,

15 Australoporus tasmanicus, Inonotus nothofagi, Pleurotus purpureo-olivaceus and an unidentified

16 hymenochaetaceous species. Richness and diversity of basidiomycete species were greater in stems of red

17 beech (Nothofagus fusca) and silver beech (N. menziesii) than in those of matai (Prumnopitys taxifolia,

18 Podocarpaceae) and tawa (Beilschmiedia tawa, Lauraceae), as determined from earlier studies in podocarp

19 hardwood and beech indigenous forests. There was greater similarity in the species composition of

20 basidiomycete fungi colonising the three beech species compared with those colonising rimu (Dacrydium

21 cupressinum, Podocarpaceae), tawa and matai. Based on observations in this study and on international

22 research on the effects of selective logging on basidiomycete biodiversity, the decision to restrict to 50% the

23 extraction of wood following storm damage in beech forests on the South Island West Coast appears to have

24 been appropriate.

25 Key words: fungal biodiversity, decomposer fungi, decay of fallen Nothofagus stems, New Zealand

26 indigenous forest ecology

27 1. Introduction

28

29 The decay of woody matter is crucial to the overall forest ecosystem. The release of carbon and minerals

30 during this process offsets the reverse actions of carbon assimilation and nutrient uptake by the living trees.

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31 Both aspects must be considered for a proper appreciation of a forest’s ecology. Balancing the carbon budget

32 requires quantification not only of carbon fixation but also of its escape during decomposition (Beets et al.

33 2008; Mason et al. 2013). The bound minerals that are also freed as the wood decays are essential

34 supplements within the forest’s nutrient cycle (Clinton et al. 1999, 2009; Buchanan et al. 2001).

35

36 The organisms responsible for wood breakdown are therefore an important element within the total forest

37 biota. However, knowledge of the decomposer populations and their ecology in New Zealand’s native forests

38 is still patchy. In particular, how is the diversity of these species affected by the creation of woody debris

39 through wind throw, and conversely, by the removal of stems during sustainable logging or in salvage

40 recovery after storms? In April, 2014, widespread damage from Cyclone Ita affected native forests in the

41 West Coast Region of the South Island (Platt et al. 2014). In order to balance conflicting commercial and

42 conservation interests, the New Zealand Department of Conservation (DOC), with assistance and advice 43 from the Ministry for Primary Industries (MPI),Draft authorised log recovery in up to 50% of the affected areas on a 44 hectare by hectare basis, and up to 50% of the fallen wood within these permitted salvage zones (Watson

45 2017). The remaining fallen stems were left as habitat shelter for bird and insect species occupying and

46 feeding in decaying logs and for nutrient recycling purposes. However, the knowledge that determined these

47 criteria appears limited. Certainly, virtually nothing is known about the way wood recovery influences the

48 residual populations of wood decay fungi in New Zealand native forests. There has been work in relatively

49 undisturbed forests, but even here more studies are needed. An awareness of these fungi and the way they

50 colonise the woody debris under ostensibly normal circumstances provides a necessary benchmark for

51 making comparisons after catastrophic weather events.

52

53 Field work to investigate the biology of fungi causing wood decay in New Zealand native forests has been

54 conducted intermittently over the last three decades, not counting the species lists of Gilmour (1966) and

55 McKenzie et al. (2000), which include ecological observations. Studies in two widely separated podocarp

56 hardwood forests, one in each of the North and South Islands, indicated a degree of commonality with

57 respect to the dominant decay fungi present in fallen stems of three tree species, rimu (Dacrydium

58 cupressinum Sol. ex Lamb., Podocarpaceae), matai (Prumnopitys taxifolia (Banks & Sol. ex D. Don) de

59 Laub., Podocarpaceae) and tawa (Beilschmiedia tawa (A. Cunn.) Kirk, Lauraceae) (Hood et al. 1989, 2004;

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60 Hood 2012; Hood and Gardner 2009). On the other hand, two studies in beech (Nothofagus, Nothofagaceae)

61 forests, one in each island, revealed considerable variety in composition between the major decomposer

62 species colonising different trees (Allen et al. 2000; Hood et al. 2008). This disparity may have been due to

63 differences in location (and climate), tree species, individual stems (varying in size and degree of

64 decomposition), supply of fungal inoculum, the study methodology used, or some combination of these or

65 additional factors. The study in the South Island monitored the fruitbodies appearing seasonally on fallen

66 mountain beech trees (Nothofagus solandri var. cliffortioides (Hook. f.) Poole; synonym, Fuscospora

67 cliffortioides (Hook. f.) Heenan & Smissen; Allen et al. 2000). The North Island study considered the fungi

68 isolated from within fallen stems of red beech (Nothofagus fusca (Hook. f.) Oerst.; synonym, Fuscospora

69 fusca (Hook. f.) Heenan & Smissen) and silver beech (Nothofagus menziesii (Hook. f.) Oerst.; synonym,

70 Lophozonia menziesii (Hook. f.) Heenan & Smissen); Hood et al. 2008).

71 72 In order to comprehend this variety more clearlyDraft further sampling is needed. A new study was therefore 73 undertaken in the South Island employing the same isolation technique as used in the central North Island

74 Nothofagus investigation. It was decided to sample all three Nothofagus species previously examined and to

75 choose sites at a number of locations. In contrast to earlier studies, which were mostly conducted following

76 known storm events, it was not possible in this study to determine the period since windfall. Instead, trees at

77 a similar, intermediate stage of physical decomposition were selected.

78

79 The purpose of the study, then, was to characterise the composition, incidence, diversity and internal

80 distribution of the fungi decaying fallen Nothofagus trees of different species at separate locations, in order to

81 add to the body of knowledge about these organisms in indigenous forests in New Zealand. As a secondary

82 aim, it was anticipated that, although conducted in comparatively undisturbed forests, it would provide

83 information on the effects that salvage logging in storm damaged stands might have on the prevalence of

84 these fungi.

85

86 2. Methods

87

88 2.1 Sites

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89

90 The study was undertaken at three South Island locations, in red and silver beech mixed forest in the Maruia

91 Valley between Springs Junction and Lewis Pass (Latitude -42.381°, Longitude 172.307°; 560m a.s.l.), in

92 mountain beech forest at Klondyke Corner near Arthur’s Pass (-43.006°, 171.582°; 640m a.s.l.), and in the

93 Eglinton Valley between Te Anau and Milford Sound. The Eglinton Valley location was subdivided into two

94 sites 11 km apart, in mountain beech forest at Boyd Creek (-45.135°, 167.950°; 320m a.s.l.) and in red and

95 silver beech forest at East Branch (-45.050°, 168.012°; 360m a.s.l.). The two most distant locations spanned

96 a length of ca. 460 km.

97

98 2.2 Sample trees

99

100 Five uprooted trees or fallen, wind-snapped stems were chosen at each location (15 trees, total; Table 1; Fig. 101 1a,b). Trees at the Maruia Valley location wereDraft of silver beech, at Klondyke Corner of mountain beech, and in 102 the Eglinton Valley of mountain beech (Boyd Creek) or red beech (East Branch). Selection was arbitrary,

103 without prior examination for any fungal fruitbodies present, except that stems were avoided that were either

104 recent falls with hard sound wood, or conversely, extensively decomposed with friable texture, collapsed form

105 and little remaining bark. Chosen trees were mostly scattered in distribution, beneath a still intact canopy,

106 over a distance that varied between 60 and 900 m at each location or site.

107

108 2.3 Field sampling

109

110 Five discs ≥ 5 cm thick were cut with a chain saw at approximately equal intervals along the available

111 unbranched length of each stem. Depending on the stem length and width, the distance between discs

112 ranged between 1.0 and 3.7 m and disc diameters varied between 16.1 and 74.5 cm (Table 1). Four mutually

113 perpendicular radial sectors were cut from each disc, one opposite pair being oriented parallel with the

114 ground surface as in the intact stem (Fig. 2a). On each sector a reference datum mark was inscribed with an

115 indelible pencil at a set radial distance in from the position of the cambium. Sectors were held individually in

116 labelled polythene bags packed into closed polystyrene boxes kept cool with icepacks changed daily for

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117 periods up to 16 days, and thereafter indefinitely in a refrigerator at 4 °C. A record was kept of locations of

118 fungal fruitbodies along stems, some of which were collected for laboratory examination.

119

120 2.4 Isolation of fungi

121

122 Isolations were undertaken in the laboratory between 6 and 10 weeks after sampling. Each sector was

123 carefully split with a small axe along the radial longitudinal plane to expose a fresh clean surface. Small chips

124 (ca. 1–2 mm across) were cut aseptically along the radius, four from the 0–6 cm depth zone, three from the

125 6–12 cm zone, and three from along the full extent of the zone deeper than 12 cm (Fig. 2b), and plated onto

126 malt extract agar (2% malt extract, Halcyon Proteins Pty. Ltd., Victoria, Australia; 1.5–2% bacteriological

127 agar, Seaweeds Division, New Zealand Manuka Group; in water). A second, identical set of 10 chips was

128 taken from the same positions and plated onto a medium selective for basidiomycetes consisting of malt 129 extract agar supplemented with 100 ppm streptomycinDraft sulphate (Thermo Fisher Scientific) and 10 ppm 130 benomyl (as 50% active ingredient commercial benlate fungicide, Du Pont). For shorter sectors from smaller

131 disks, chips were taken as described only from available outer depth zones. Plates were sealed, placed in

132 polythene bags stored in a closed, opaque, white, plastic box, incubated at ca. 20 °C under ambient room

133 lighting, and monitored for periods of up to 4 months. Emerging isolates were cut out and sub-cultured

134 individually in tubes of 2% malt agar. Where identical isolates emerged in larger numbers, an arbitrary

135 subset, only, was sub-cultured (Fig. 1c,d).

136

137 2.5 Identification of fungi

138

139 Selected representative isolates were identified as precisely as possible morphologically and by molecular

140 sequencing. Both procedures were complementary, with the emphasis being weighted towards one or other

141 approach according to degree of familiarity with isolate attributes. Morphological identification was

142 undertaken by growing cultures for six weeks on plates of 2% malt extract agar using the methods of Nobles

143 (1965) and Stalpers (1978) as previously modified (Hood et al. 1989) to determine growth rates and

144 characteristic macro- and microscopic characters. Comparisons were made with matching culture

145 descriptions of isolates made from identified fruit bodies during earlier work and between isolates from

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146 different discs and trees in the present study. Non-sporulating cultures not confirmed as basidiomycetes (e.g.

147 lacking clamp connections or not recognised as a known species) were treated with α naphthol to indicate

148 presence of laccase and hence likely white rot capability (Stalpers 1978). The study focused on wood decay

149 fungi and only minimal attempt was made to identify other types.

150

151 DNA of representative isolates of more common species was sequenced in the ITS (internal transcribed

152 spacer) region. Cultures were grown on 2% malt agar with cellophane at 20C for 5–7 days. DNA was

153 extracted using a FastDNA kit (MP Biomedicals, OH, USA) according to the manufacturer’s instructions.

154 Mycelium was scraped into lysing matrix A tubes with cell lysis solution CLS-Y and DNA was purified

155 following the manufacturer’s instructions. DNA integrity was checked by agarose gel electrophoresis with

156 1.5% (wt./vol.) agarose in TBE (150 V for 35 min), stained with RedSafe™ (iNtRON Biotechnology, WA,

157 USA) and visualised by UV transillumination. DNA was quantified using Denovix® DS-11 (DeNovix Inc., NC,

158 USA) and kept at -20C for long term storage.Draft

159

160 PCRs with ITS4 and ITS1F primers (Gardes and Brun 1993) were performed using the HOT FIREPol® Blend

161 Master Mix (5X) (Solis BioDyne, Tartu, Estonia), according to the manufacturer’s instructions. Each 20 µl

162 PCR reaction contained 7.75 µl of PCR grade water, 4.0 µl 5X HOT FIREPol® Blend Master Mix, 2 mM MgCl2,

163 300 nM of each primer, 2 µL DNA (approximately 5–10 ng gDNA). The cycling conditions consisted of an

164 initial denaturation step of 94 ºC for 15min, then 35 cycles of 94 ºC (30s), annealing at 55 ºC (40s), and 72ºC

165 (1min), and a final extension step of 72ºC (10 min). Gel electrophoresis was performed with 1.5% (wt./vol.)

166 agarose in TBE (150 V, 25 min), stained with RedSafe™ (iNtRON Biotechnology, WA, USA) and visualised

167 by UV transillumination.

168

169 Prior to DNA sequencing, PCR products were treated with Exonuclease I (Exo) and Fast Alkaline

170 Phosphatase (FastAP) (Fermentas, Thermo Fischer Scientific, Waltham, MA, USA) according to the

171 manufacturer’s instructions. DNA sequencing was performed in both directions with ITS1F and ITS4 target

172 primers. DNA sequencing was performed on a 3500 Genetic Analyzer machine (Applied Biosystems), using

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173 BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems), according to the manufacturer’s

174 instructions.

175

176 DNA sequences were analysed in Geneious 10.2.2 (Biomatters Ltd., New Zealand) (Kearse et al. 2012).

177 Blast searches of the National Centre for Biotechnology Information (NCBI) GenBank database

178 (https://www.ncbi.nlm.nih.gov/genbank/) were undertaken to help identify sequenced isolates to species or to

179 a broader taxonomic grouping, where possible. An identification to species was accepted only if there was ≥

180 (97–) 99% sequence equivalence to a reliably identified GenBank item, with a high base pair percentage

181 query cover and an E value  8 × 10-70. DNA sequences were aligned using MAFTT v1.3.6 (Katoh et al.

182 2002) and phylogenetic analyses were performed with the Geneious plugin, RAxML v7.2.8, applying the

183 default model parameters of GTR GAMMA, with 100 bootstrap replicates (Liu et al. 2011). These analyses

184 were performed within the Geneious software.

185 Draft

186 2.6 Data analysis

187

188 Generalised linear mixed models (GLMMs) were used to determine whether the percentage of isolation

189 attempts yielding fungi (percentage yield; both isolation media combined) varied between host species,

190 location and radial depth. Models were fitted for all fungi and just basidiomycetes. Each model included

191 location by host species (Maruia Valley/silver beech; Klondyke Corner/mountain beech; Eglinton

192 Valley/mountain beech; Eglinton Valley/red beech) and radial depth zone (0–6, 6–12, >12 cm) as fixed

193 effects. Stem, nested in species by location, was included as a random effect. The dependent variable was

194 percentage yield fitted using binomial GLMMs (assuming a binomial error and logit link). Overdispersion

195 occurred and was modelled using a quasi-binomial GLMM (Zuur et al. 2013). The model parameters were

196 estimated using penalised quasi-likelihood (PQL). The Tukey HSD method was applied as a post-hoc test to

197 conduct pairwise comparisons between means. The model was fitted using the R-MASS package.

198

199 Species richness and diversity of fungal communities were compared among 54 fallen stems of six

200 indigenous host species by combining raw data from this and four earlier studies that used a comparable

201 sampling technique (Hood et al. 2004, 2008; Hood and Gardner 2009; Hood 2012). The host species were

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202 rimu (two locations: North Island, 8 stems; South Island, 5 stems; mean and standard deviation of stem disc

203 diameter means, 61.0 ± 14.4 cm), matai (one location: North Island, 8 stems; 54.9 ± 14.0 cm), tawa (one

204 location: North Island, 6 stems; 37.8 ± 9.9 cm), red beech (two locations: North Island, 6 stems; South Island,

205 present study, 2 stems; 43.3 ± 15.8 cm), silver beech (two locations: North Island, 6 stems; South Island,

206 present study, 5 stems; 38.8 ± 9.4 cm) and mountain beech (two locations, South Island, present study, 5

207 and 3 stems; 25.6 ± 5.3 cm). Disc diameters for the South Island rimu location were estimated from the basal

208 breast height disc measurement and a standard rimu stem taper table for Westland, all other discs being

209 directly measured.

210

211 Richness and diversity indices were calculated using Hill numbers of orders 0, 1 and 2, with the order

212 indicating the weighting assigned to common versus rare species (Hill 1973). The Hill number of order 0 is

213 simply species richness or number of species which assigns an equal weighting to all species. The Hill 214 number of order 1 is the exponential of Shannon’sDraft index (Shannon 1948) which assigns a lower weighting to 215 rare species, while the Hill number of order 2 is Simpson’s inverse index (Simpson 1949) which assigns an

216 even lower weighting to rare species. All three indices increase in value with increasing diversity. Hill

217 numbers were calculated using the methods given for incidence data by Chao et al. (2014). These are as

218 follows. If pi is the proportion of positive isolation attempts yielding species i within a stem (isolation attempts

219 yielding no fungi are ignored), then the Hill numbers are:

0 0 220 Hill number of order 0: 퐷 = ∑푝푖

1 221 Hill number of order 1: 퐷 = 푒푥푝( ― ∑푝푖푙푛(푝푖))

2 1 2 222 Hill number of order 2: 퐷 = ∑푝푖

223 Although similar sampling methods were used in all studies, the average number of isolation attempts per

224 stem varied between studies, ranging between 114 for tawa to 475 for matai. Because empirical estimates of

225 diversity tend to increase with sampling effort (Chao et al. 2014), it was necessary to standardise the

226 estimates for sampling intensity. This was achieved by randomly sampling results of 120 isolation attempts

227 from each stem and discarding the remainder. Although this method meant that only a portion of the available

228 data was used, very similar results were obtained when the process was repeated several times using

229 different random selections of samples, indicating that the results obtained were robust. For completeness,

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230 results using all available data were also determined, although in this case comparisons between species

231 were confounded by the varying levels of sampling intensity used in different studies.

232

233 Calculations of the richness and diversity indices were restricted to yield data obtained using only the

234 selective isolation medium, which was common to all studies. Isolates designated in general terms only

235 (basidiomycete sp., other hymenochaetaceae sp.) were grouped collectively as single “species”; those of the

236 genera Armillaria and Sistotrema, where the precise species identity was not known for all isolates, were

237 likewise grouped conservatively as single “species”; laccase positive isolates, not all of which were

238 necessarily basidiomycetes, were excluded from the index calculations. Species richness and diversity

239 indices were compared between host species by 1-way analysis of variance (GLM procedure, SAS Version

240 9.4). All combinations of species and location were also compared by ANOVA. All diversity indices were log

241 transformed before these analyses. The Tukey HSD method was used for pairwise comparisons between 242 means. Draft 243

244 The data obtained from the 54 fallen stems in this study and four earlier studies were also used to analyse

245 the species composition of basidiomycete fungi among the six indigenous host species. In this analysis the

246 data used consisted of the presence/absence of each of the 51 basidiomycete species listed for each host

247 species. Dissimilarity in species composition between each pair of host species was quantified using the

248 Jaccard dissimilarity coefficient calculated from these presence/absence data (DISTANCE procedure of SAS

249 Version 9.4). If NA is the number of basidiomycete species on host A, NB is the number of basidiomycete

250 species on host B, and NAB is the number of species common to both A and B, then the Jaccard dissimilarity

251 index is calculated using:

푁 252 푑퐽 = 1 ― 퐴퐵 (푁퐴 + 푁퐵 ― 푁퐴퐵)

253 A cluster analysis was then performed to identify similarity in fungal communities between host species

254 (CLUSTER procedure of SAS Version 9.4 using average linkage clustering).

255

256 A corresponding “countrywide” analysis of basidiomycete species composition among the six indigenous host

257 species was undertaken using data on a total of 475 species of wood colonising basidiomycetes obtained

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258 from collections held in two mycological herbaria, PDD (New Zealand Fungal Herbarium, Landcare

259 Research, Auckland, New Zealand; https://nzfungi2.landcareresearch.co.nz/) and NZFRI-M (New Zealand

260 Forest Research Institute Mycology Herbarium, Rotorua, New Zealand). The presence/absence of each

261 basidiomycete species for each host species was used to calculate dissimilarities in composition between

262 pairs of host species and then perform a cluster analysis using the same procedure as outlined above.

263

264 3. Results

265

266 A total of 47% of 4797 isolation attempts yielded cultures of different fungi (Table 2). Basidiomycete species

267 were obtained overall from 30% of attempts, or from 38% of attempts on the basidiomycete selective medium

268 and from 22% of those on straight malt agar. Conversely, 2.4% of attempts on malt agar, but only 0.4% of 269 attempts on the selective medium, yielded culturesDraft of ascomycete fungi in the Xylariales (Table 2). Stems of 270 all host species were well colonised at all sites and radial depths and there were few trends (Tables 3 and 4).

271 However, when taken together, slightly fewer isolates of all fungi were obtained from depths greater than 12

272 cm than from nearer the surface (Table 4). With respect to basidiomycetes alone, the same trend of fewer

273 isolates at depths below 12 cm was apparent only in mountain beech (N. solandri var. cliffortioides) at

274 Klondyke Corner (Table 4).

275

276 Of the fungi isolated a minimum of 29 basidiomycete species and three ascomycete species in the Order

277 Xylariales were recognised (Table 5). Identification ranged from complete, with precise species identity, to

278 partial, within a broader taxonomic grouping only, according to current morphological and molecular

279 knowledge (App. A; brief culture descriptions are provided as Supplementary Material, Table S1). The

280 basidiomycetes most frequently isolated were (Table 5) Australoporus tasmanicus (At; 11% of all

281 basidiomycete isolates; 3.0% of isolation attempts, meaned over the three locations), Ganoderma sp. (G;

282 19%; 5.3%), an unidentified hymenochaetaceous species (H4; 17%; 5.4%), Inonotus nothofagi (In; 11%;

283 3.0%) and Pleurotus purpureo-olivaceus (Pp; 18%, 5.8%). Eleven species produced fruitbodies or other

284 fungal signs on one or more stems (Table 6). These included A. tasmanicus (At), G. applanatum sensu

285 Wakefield (Ga; the only species of this genus confirmed from examination of fruitbodies as present in the

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286 study), C. tabacinus (Ct), and species of Armillaria (A) and Biscogniauxia (Bc, Br). Rhizomorphs or mycelial

287 fans of Armillaria sp(p). were present in three stems.

288

289 A pictorial, three-dimensional representation of the distribution of basidiomycete and ascomycete species

290 within each of the 15 stems is depicted in Figs. 3–5, based on isolation and occurrence of fruitbodies and

291 other fungal signs. A number of features are indicated. The more common species occupied significant tracts

292 of stem (two or more adjacent discs). These included Ganoderma sp. (G; stems 1, 4, 5, 6, 9); A. tasmanicus

293 (At; stems 1, 5, 14, 15); an unidentified hymenochaetaceous species (H4; stems 14, 15); I. nothofagi (In;

294 stems 2, 8); and P. purpureo-olivaceus (Pp; complete 4 m sampled length of stem 13). Other species

295 showing the same trend included an unidentified hymenochaetaceous species (H2; stems 3, 4); Postia

296 pelliculosa (Ppe; stem 10); Schizopora radula (Sr; stem 12); ascomycete Biscogniauxia sp. (B; stem 6);

297 ascomycete Diatrypella sp. (D; stem 7); and unidentified xylariaceous ascomycete (Xy; stem 12). Armillaria 298 novae-zelandiae (An; stem 8) may have shownDraft the same pattern (only one Armillaria isolate from this tree 299 was identified to species by sequencing).

300

301 Fruiting occurred at or near the region of isolation of the same species (Figs. 3–5) for G. applanatum sensu

302 Wakefield (Ga, G; stem 1, discs 1–2; stem 4, near disc 1; stem 6, between discs 1–3) and A. tasmanicus (At;

303 stem 14, discs 1–3; stem 15, discs 4–5). Fruiting of Phellinus kamahi (Pk) and a thick yellow padded

304 mycelium coincided with the zone yielding unidentified hymenochaetaceous species H2 in stem 3. Pleurotus

305 purpureo-olivaceus (Pp) was associated with vigorous, white, padded mycelium in three stems (stem 11,

306 discs 2, 3; stem 13, discs 2–5; stem 15, disc 3). Postia pelliculosa (Ppe) was isolated from a brown wood rot

307 (stem 10, disc 4) and Ganoderma sp. (G) from a soft white decay (stem 1, discs 1, 2; stem 4, discs 1, 2).

308 Characteristic white mycelial fans were present beneath the bark on disc 5 (stem 8) from which A. novae-

309 zelandiae was obtained. Nevertheless, isolates were not obtained of all species that fruited on study stems

310 (Table 6). These included Fomitopsis hemitephra (Fh; stem 1, discs 3–4), Stereum hirsutum (Sh; stem 1,

311 discs 4–5), Hohenbuehelia luteohinnulea (Hol; stem 12, disc 5), and T. versicolor (Tv; near disc 1, stems 12

312 and 15). Two varieties of B. capnodes (Bc, Br) were observed fruiting on six stems (Table 6; stem 1, disc 1;

313 stem 5, disc 3, stem 6, disc 4, stem 7, disc 1; stem 12 , disc 4; stem 15). However, isolates of Biscognauxia

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314 sp. (B; stem 6, discs 4, 5; stem 7, disc 5; stem 12, discs 2, 3) appeared to be of a different species (App. A,

315 footnote 3).

316

317 A total of 16 species of basidiomycete and ascomycete (Xylariales) fungi were recorded as occurring on more

318 than one stem (Table 7). Of these, seven species were widely distributed by both host and location,

319 comprising A. novae-zelandiae (An), A. tasmanicus (At), B. capnodes (both varieties; Bc, Br), F. hemitephra

320 (Fh), Ganoderma sp. (G), I. nothofagi (In) and an unidentified species that may belong to Gymnopus (Gy).

321 Seven species were present on stems of one host species only at one site (Table 7). These were on

322 mountain beech: P. purpureo-olivaceus (Pp), T. versicolor (Tv), and a crepidotaceous (Cr1) and

323 hymenochaetaceous species (H4); on silver beech: another hymenochaetaceous species (H2); and on red

324 beech: C. tabacinus (Ct) and P. pelliculosa (Ppe). Xylariaceous species (Xy) was also present only on

325 mountain beech, but at two sites (Table 7). 326 Draft 327 Basidiomycete fungi varied significantly in both richness and diversity in fallen stems among host species

328 when results from this and four other similar studies were examined together (Table 8; Supplementary

329 Material, Table S2). Generally, all three indices showed highest richness and diversity in stems of red beech

330 (N. fusca) and silver beech (N. menziesii), intermediate diversity in those of rimu (D. cupressinum) and

331 mountain beech (N. solandri var. cliffortioides), and lowest diversity in tawa (B. tawa) and matai (P. taxifolia)

332 stems.

333

334 The composition of basidiomycete fungi also varied between the six indigenous hosts both in the fallen stems

335 in the present and earlier studies and, at the broader countrywide level, among the collections of wood

336 colonising basidiomycetes held in national mycological herbaria (Supplementary Material, Table S3, Table

337 S4). Although many basidiomycete species were shared in common among all six indigenous tree species

338 (Table S3), others were more limited in their host range. Disregarding matai (P. taxifolia), clustering of host

339 species based on Jaccard similarities showed a parallel pattern in both the study trees and also among the

340 herbarium records (Supplementary Material, Fig. S1). In each data set, the three beech (Nothofagus) species

341 clustered separately from the two tree species (rimu and tawa) in podocarp hardwood forest. Matai also

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342 clustered with rimu in the broader, countrywide data set, but among the study trees it separated out on its

343 own, although was closer to rimu than any of the other host species (Fig. S1).

344

345 4. Discussion

346

347 The work described in this paper extends the findings of earlier field studies and increases our understanding

348 of the biology of wood decay fungi in native forests in New Zealand. More generally, it contributes to the

349 wider body of knowledge of this group internationally, particularly in the southern hemisphere, where a

350 number of basidiomycete species occur that are not present north of the equator. The results of field

351 investigations also complement those of several experimental studies investigating the relative

352 competitiveness and ability to decompose wood of some of the same and other fungal species (Buchanan et

353 al. 2001; Clinton et al. 2009; Fukami et al. 2010; Dickie et al. 2012; cf. Beets et al. 2008). 354 Draft 355 The nature of the fungal community within a fallen tree stem is defined by its composition (the particular

356 fungal species present), richness (their number), and diversity (the relative abundance of each of those

357 species). Diversity is an expression of the degree to which a stem is occupied internally by numerous small

358 colonies of many different fungi or by extensive, vigorous, vegetative mycelia of a few predominant species

359 (for instance, P. purpureo-olivaceus in stem 13). In this study basidiomycete diversities were calculated from

360 isolation yields using two indices that gave mutually equivalent results, which also matched the richness

361 determinations. For a more precise measure of diversity it will ultimately be necessary to allow for some

362 single species being present in the form of numerous, contiguous, unique dikaryons or individuals belonging

363 to discrete vegetative compatibility groups as a result of multiple basidiospore colonisations from a nearby

364 inoculum source or spore rain, as is known to occur with fungi such as G. applanatum sensu Wakefield,

365 Phlebiopsis gigantea (Fr.) Jülich and T. versicolor: Todd and Rayner 1978; Hood et al. 2004, 2015). Even

366 the vegetative compatibility group may not be the ultimate unit of the individual genotype for some species,

367 such as Armillaria limonea (G. Stev.) Boesw. and A. novae-zelandiae (Hood 2012). In the present

368 investigation the character of the wood decay communities (their composition, richness, diversity in the form

369 derived, and three dimensional internal distribution) was determined in fallen stems of three Nothofagus

370 species in indigenous beech forest. When compared with results from previous studies, basidiomycete

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371 communities were richer and more diverse within fallen beech stems than in stems of two conifer (rimu,

372 matai) and one hardwood (tawa) tree species in podocarp hardwood forest. At a higher level, among all 54

373 stems, the composition of basidiomycete species also differed between the beech species and the tree

374 species in podocarp hardwood forests. In terms of both numbers of stems colonised and incidence of

375 isolation, species of Armillaria and Ganoderma were unequivocally the dominant basidiomycetes in podocarp

376 hardwood forests at geographically distant sites (Hood et al. 2004; Hood and Gardner 2009; Hood 2012).

377 Armillaria and Ganoderma species were also important in fallen beech wood, but were accompanied by a

378 broader range of other basidiomycete species, which differed in composition between stems at the same and

379 separate locations. For instance, C. tabacinus, which was prominent in both red and silver beech stems in the

380 earlier central North Island study (Hood et al. 2008), was isolated only infrequently in the current

381 investigation, with other species present instead. Armillaria species are responsible for a root disease that

382 once afflicted exotic Pinus radiata D. Don stands planted on land cleared of indigenous forest, particularly on 383 ex cutover podocarp hardwood sites (ShawDraft and Calderon 1977; Hood and Sandberg 1987, 1993). 384

385 Composition, richness and diversity of wood decay fungi are governed by many factors both external and

386 intrinsic. Some basidiomycetes are inherently more vigorous and able to outcompete others, and species

387 may naturally employ different occupation strategies along the lines of the ruderal versus seral approaches of

388 higher plants. Basidiomycetes are generally able to colonise stems of a broad range of hosts, but certain

389 related plant groups (e.g. angiosperms, gymnosperms, or members of a common genus or family) may differ

390 in their susceptibility to particular species for reasons not entirely clear, except that they presumably relate to

391 particular properties of heartwood or sapwood. Wood rot fungi differ innately in their ability to produce

392 enzymes capable of denaturing cellulose and lignin. The difference in the composition of basidiomycete

393 species within stems between host species in beech and podocarp forests in the study trees was mirrored at

394 the countrywide level in the data from national mycological herbaria, implying that it was part of a broader

395 natural pattern. The reason for the one exception to this in stems of matai (P. taxifolia) is not obvious. Matai

396 stems consist mostly of a cylinder of decay resistant heartwood, with only a narrow outer ring of susceptible

397 sapwood, and the basidiomycete species recorded from this host in national herbaria were comparatively low

398 in number (Supplementary Material, Table S3(b)). It is noteworthy that the composition of basidiomycete

399 fungi in fallen stems in both types of indigenous forest was quite distinct from that in stumps in New Zealand

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400 P. radiata plantations where at least some species are also found in northern hemisphere pine stands and

401 which like their host may be introduced (Hood et al. 2015).

402

403 Externally, composition, richness and diversity are influenced by variables such as environmental conditions,

404 inoculum supply, possibly the physical position of stem entry, and the volume of substrate available for

405 colonisation (Fryar et al. 1999). In this study, richness of basidiomycete species appeared lower (numerically

406 if not significantly) in the mountain beech (N. solandri var. cliffortioides) stems than in red (N. fusca) and silver

407 (N. menziesii) beech stems sampled in the present and previous studies, which could potentially reflect their

408 smaller size. Allen et al. (2000) reported that richness of fungal taxa was lower in stems of lesser volume,

409 based on fruitbody collections from 75 mountain beech stems. Their overall mean values of 3.6 (spring) and

410 6.0 (autumn) for a broad grouping of different macrofungi (Allen et al. 2000) compares with 3.1 species per

411 stem for basidiomycetes, only, among the eight mountain beech stems in this study. A lower richness in 412 smaller stems may also be due to factors otherDraft than simply a reduced volume of substrate. It is possible that 413 the occurrence of some fungi may be restricted if environmental conditions are less than favourable for

414 growth (for instance, if wood moisture content, not measured in this study, is critically low in smaller stems; cf.

415 stem 11, Fig. 5, although overall basidiomycete yield was not noticeably lower in the smaller mountain beech

416 stems, Table 4). At the countrywide level, richness of wood colonising basidiomycete species reported in

417 mountain beech (150 species) is not less than in red beech (130 species; McKenzie et al. 2000; cf.

418 Supplementary Material, Table S3(b)).

419

420 Identification of species in this study was based on traditional culture morphology, by comparing isolates from

421 fallen stems with others made from confirmed fruitbodies, and on molecular sequencing of isolates. Equating

422 sequences to those held on international databases such as GenBank is invaluable but currently hampered

423 by the uneven representation of authentic reference sequences (Vaz et al. 2017), especially for species in the

424 southern hemisphere. A number of species isolated could be assigned only to a broad taxonomic group

425 (representative cultures have been deposited in collection NZFS in anticipation of their eventual precise

426 identification). Nevertheless, the situation is improving as more items are added and it was gratifying to be

427 able to identify a number of species, such as P. purpureo-olivaceus, only by this means, because of previous

428 work by others. It is also apparent that although isolation gave a more complete picture, a full understanding

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429 of the fungal communities within stems also requires monitoring of fruitbodies. In this study colonisation by

430 some species was recognised only by the production of fruitbodies, while for a number of others that were

431 isolated, fruitbodies were not seen. This was especially, though not exclusively true of species whose

432 fructifications are ephemeral and seasonal. Isolation of Biscogniauxia species seemed low in relation to

433 observed fruiting. Even using both approaches, it is likely that some less common or not readily culturable

434 species will still not have been detected, despite the intensity of isolation in this study, which made it

435 impractical to sample more than a relatively limited number of stems. Current international research is

436 addressing this issue in much greater detail by means of high capacity next generation sequencing

437 approaches using environmental DNA (Kubartová et al. 2012; Purahong et al. 2018; Bulman et al. 2018),

438 recognising that these methods also rely on the quality and completeness of reference sequence databases

439 and do not distinguish actively functioning mycelia from spores or (unlike culturing techniques) dead fungal

440 material. 441 Draft 442 An interesting circumstance was the presence in fallen wood of fungal species that have also been recorded

443 as endophytes in foliage. Xylariaceae sp. 4, Biscogniauxia sp. 1 and Biscogniauxia sp. 2 of Johnston et al.

444 (2012), from symptomless leaves of Nothofagus species, were all identified by sequencing in this study, the

445 first two from wood and the last from fruitbodies of B. capnodes (App. 1, footnotes 3, 6). In a similar way,

446 basidiomycete isolates from felled, exotic Pinus radiata stems were identified by a sequencing match to an

447 endophytic Cylindrobasidium sp. from foliage of the indigenous podocarp Dacrydium cupressinum (Joshee et

448 al. 2009; McCarthy et al. 2012). It has been demonstrated that some foliar endophytes may confer an

449 advantage by protecting their hosts against drought, pathogens, insect browsers or mammalian herbivores

450 (Carroll 1995; Bullington and Larkin 2015). Equally, foliage may serve as a refuge for wood colonising fungi if

451 substrate is not available or when conditions are not suitable for establishment (Carroll 1999; Martin et al.

452 2015; Thomas et al. 2016). This could be relevant to the local survival of some species when wood is

453 removed during salvage logging following storms (see below).

454

455 In this study, seven basidiomycete decay species were identified by isolation or fruiting in either two or three

456 of five stems in the same vicinity at only one site. This clustered distribution appears noteworthy even though

457 numerically small. Disregarding the possibility of host specificity for some species, such a localised

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458 distribution pattern is suggestive of colonisation of new wood by comparatively short distant dispersal of local

459 basidiospore inoculum. The disjunct upper ends of stems 1 and 5 from the same tree may also have been

460 colonised separately by locally dispersed basidiospores of A. tasmanicus (Fig. 3). Spatially limited spore

461 dissemination and colonisation from a nearby inoculum source has previously been suggested with species

462 of Armillaria, Ganoderma and Kretzschmaria zelandica J.D. Rogers & Y.M. Ju (Hood and Sandberg 1993;

463 Hood et al. 2004; Power et al. 2008; Hood and Gardner 2009; not discounting more distant travel by a

464 minority of the spores: Stenlid and Gustafsson 2001). These observations conform to experimental work

465 demonstrating that the composition of fungal communities within woody material in forests depends to a large

466 extent on the order in which inoculum of different fungi first arrives at and occupies the fresh substrate

467 (Fukami et al. 2010; Dickie et al. 2012), notwithstanding the potential eventual replacement of some species

468 through competition (Stenlid and Gustafsson 2001; Jönsson et al. 2008). Detailed spore trapping studies with

469 a number of basidiomycete species have demonstrated and quantified a restricted and uneven inoculum 470 dispersal, with the implication that for rare speciesDraft removal of woody material or fragmentation of tracts of 471 forest may diminish their occurrence (Norros et al. 2012; Peay and Bruns 2014). Research into the effects of

472 removing wood substrate from forests has generally indicated a reduction in the biodiversity of wood

473 colonising basidiomycete fungi (Bader et al. 1995; Sippola et al. 2001; Penttilä et al. 2004; Müller et al. 2007;

474 Hattori et al. 2012; Tchoumi et al. 2017). For common and well dispersed wood inhabiting fungi in New

475 Zealand there seems little chance that salvage logging after storms will affect their wider distribution and

476 survival. However, for less common, locally distributed, or rare species

477 (https://www.landcareresearch.co.nz/science/portfolios/defining-land-biota/fungi/rare-and-endangered-fungi;

478 accessed 23 June, 2018) it does appear that extraction of wood may reduce their numbers and possibly

479 threaten their existence. In balancing various external pressures following Cyclone Ita, and in the absence of

480 further information, it seems that, for wood colonising fungi, the decision by DOC to leave material in the

481 forest during log recovery was appropriate.

482

483 5. Acknowledgements

484

485 Thanks are due to DOC for support funding and to Scion for access to facilities and in-kind laboratory

486 assistance. Additional finance was provided from the Strategic Science Investment Fund by the New Zealand

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487 Ministry of Business, Innovation and Employment. Technical help was given by Sara Carey, Liz Cunningham,

488 Rachel Hood, Tomoko Pearson, Pam Taylor and Rita Tetenburg. Greg Scott prepared Figs. 3–5. Peter

489 Johnston and Adrienne Stanton are thanked for the loan of material from Herbarium PDD (New Zealand

490 Fungal Herbarium, Landcare Research, Auckland). Help in organizing access to field sites was provided by

491 Tim Shaw and Jane Marshall (DOC, Hokitika), Ben Hodgson (DOC, Greymouth), Chris Stewart (DOC, Arthur’s

492 Pass), and George Ledgard (DOC, Te Anau). The authors also thank two anonymous reviewers for helpful

493 comments that significantly improved the manuscript.

494

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666 Stalpers, J.A. 1978. Identification of wood-inhabiting Aphyllophorales in pure culture. Centraalbureau voor

667 Schimmelcultures, Utrecht, the Netherlands. Stud. Mycol. 16.

668

669 Tchoumi, J.M.T., Coetzee, M.P.A., Vivas, M.,Draft Rajchenberg, M., and Roux, J. 2017. Wood-rotting

670 basidiomycetes associated with declining native trees in timber-harvesting compartments of the Garden

671 Route National Park of South Africa. Austral Ecol. doi:10.1111/aec.12524.

672

673 Todd, N.K., and Rayner, A.D.M. 1978. Genetic structure of a natural population of Coriolus versicolor (L. ex

674 Fr.) Quél. Genetic Research 32: 55-65.

675

676 Thomas, D.C., Vandegrift, R., Ludden, A., Carroll, G.C., and Roy, B.A. 2016. Ecology of the fungal genus

677 Xylaria in a tropical cloud forest. Biotropica 48: 381–393. doi: 10.1111/btp.12273.

678

679 Vaz, A.B.M., Fonseca, P.L.C., Leite, L.R., Badotti, F., Salim, A.C.M., Araujo, F.M.G., Cuadros-Orellana, S.,

680 Duarte, A.A., Rosa, C.A., Oliveira, G., and Góes-Neto, A. 2017. Using Next-Generation Sequencing (NGS) to

681 uncover diversity of wood-decaying fungi in neotropical Atlantic forests. Phytotaxa 295 (1): 1–21.

682 doi.org/10.11646/phytotaxa.295.1.1.

683

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684 Watson, J. 2017. The recovery of windblown timber from West Coast public conservation land. N. Z. J. For.

685 62 (2): 13–16.

686

687 Zuur, A.F., Hilbe, J.M., and Ieno, E.N. 2013. A beginner’s guide to GLM and GLMM with R. A frequentists

688 and Bayesian perspective for ecologists. Highland Statistics Limited, Newburgh, Aberdeenshire, Scotland,

689 UK.

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690 Figure 1. (a), (b). Part lengths of sampled trees. (a) tree 9 (red beech, Eglinton East Branch) and

691 (b) tree 11 (mountain beech, Klondyke Corner) showing two discs. (c), (d). Cultures emerging from

692 seven chips from one sector, each (comprising 0–6 and 6–12 cm radial depth zones) of (c) Pleurotus

693 purpureo-olivaceus (ex tree 13) and (d) an unidentified hymenochaetaceous species (H4; ex tree 15);

694 both from mountain beech, Klondyke Corner.

695

696 Figure 2. (a). Positions of four radial sector samples from one disc, as looking up the fallen stem.

697 (b). Loci of isolation attempts (dots; numbered 1–10 from exterior during plating) after splitting each

698 sector sample along the radial longitudinal plane (distances from the exterior cambium surface, 0 cm,

699 divide the sector into three radial depth zones).

700

701 Figure 3. Spatial locations of decay fungi isolated from discs cut from five fallen stems (trees 1–5) of 702 Nothofagus menziesii at the Maruia ValleyDraft site (diagrammatic; vertical orientation as in field, discs 1–5 703 shown as looking up the stem, from left to right; radial depth zones from cambium of 0–6 cm, 6–12 cm

704 and >12 cm, where present, depicted as concentric circles). Fungal identity code letters, each

705 representing one or more isolates of a species, are as presented in Table 5 and App. A. Locations of

706 fruitbodies and other fungal signs are also shown, coded as in Table 6. Stems 1 and 5 originated from

707 the same multiple leader tree.

708

709 Figure 4. Spatial locations of decay fungi isolated from discs cut from three fallen stems (trees 6–8) of

710 Nothofagus solandri var. cliffortiodes at the Boyd Creek site, and from two fallen stems (9–10) of

711 Nothofagus fusca at the East Eglinton Branch site (diagrammatic; vertical orientation as in field, discs

712 1–5 shown as looking up the stem, from left to right; radial depth zones from cambium of 0–6 cm, 6–12

713 cm and >12 cm, where present, depicted as concentric circles). Fungal identity code letters, each

714 representing one or more isolates of a species, are as presented in Table 5 and App. A. Locations of

715 fruitbodies and other fungal signs are also shown, coded as in Table 6.

716

717 Figure 5. Spatial locations of decay fungi isolated from discs cut from five fallen stems (trees 11–15) of

718 Nothofagus solandri var. cliffortioides at the Klondyke Corner site (diagrammatic; vertical orientation as

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719 in field, discs 1–5 shown as looking up the stem, from left to right; radial depth zones from cambium of

720 0–6 cm, 6–12 cm and >12 cm, where present, depicted as concentric circles). Fungal identity code

721 letters, each representing one or more isolates of a species, are as presented in Table 5 and App. A.

722 Locations of fruitbodies and other fungal signs are also shown, coded as in Table 6.

723

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724 Table 1. Details of fallen Nothofagus stems sampled.

725 Site Species Stem Coordinates Fall Disc details No. (Latitude S / type Mean diameter Position on stem (m) Longitude E) (and range) Length Mean distance (cm) sampled1 (and range) between discs Maruia N. menziesii 12 -42.380012 Break (23.5-) 29.6 (-38.5) 9.2 (2.3-) 2.3 (-2.3) Valley 172.312150 2 -42.381038 Uproot (26.9-) 36.4 (-40.5) 6.4 (1.6-) 1.6 (-1.6) 172.307535 3 -42.381680 Uproot (26.0-) 37.0 (-42.5) 10.1 (2.3-) 2.5 (-2.6) 172.302086 4 -42.381825 Break (37.5-) 41.7 (-45.3) 5.8 (1.4-) 1.5 (-1.5) 172.302315 52 -42.380128 Break (25.0-) 30.4 (-39.0) 5.2 (1.1-) 1.3 (-1.5) 172.312101 Eglinton N. solandri 6 -45.134527 Break (23.4-) 30.7 (-36.0) 6.4 (1.3-) 1.6 (-1.9) Valley: var. 167.949156 Boyd cliffortioides 7 -45.134478 Break (16.4-) 18.6 (-20.1) 4.0 (1.0-) 1.0 (-1.0) Creek 167.949669 8 -45.134645 Break (23.3-) 27.2 (-34.6) 6.8 (1.7-) 1.7 (-1.7) 167.949566 Eglinton N. fusca 9 -45.049824 Break (55.6-) 62.8 (-74.5) 6.9 (1.2-) 1.7 (-2.2) Valley: 168.012397 East 10 -45.049928 DraftUproot (45.3-) 52.8 (-59.9) 12.0 (3.0-) 3.0 (-3.0) Branch 168.011702 Klondyke N. solandri 11 -43.007444 Break (24.0-) 33.5 (-37.3) 9.7 (1.8-) 2.4 (-3.7) Corner var. 171.579052 cliffortioides 12 -43.007393 Break (16.1-) 17.4 (-18.1) 4.0 (1.0-) 1.0 (-1.0) 171.579298 13 -43.005326 Break (21.0-) 27.9 (-33.4) 4.0 (1.0-) 1.0 (-1.0) 171.584781 14 -43.003316 Break (25.2-) 26.7 (-28.3) 5.1 (1.0-) 1.3 (-2.0) 171.588226 15 -43.006940 Break (19.1-) 22.6 (-25.2) 5.7 (1.2-) 1.4 (-1.6) 171.580536 726 1Distance between discs 1 and 5. 727 2Stems 1 and 5 from the same multi-leadered tree. 728 729

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730 Table 2. Percentages of all isolation attempts yielding groups of fungi by isolation medium.

731 Type of fungus Malt extract Basidiomycete Combined agar selective medium Basidiomycetes 22.1 37.7 29.9 Ascomycetes (Xylariales) 2.4 0.4 1.4 Unidentified laccase positive 2.4 1.3 1.8 Others1 21.7 5.7 13.6 Total 48.5 45.1 46.8 Total No. isolation attempts 2393 2404 4797 732 1Including hyphomycetes (e.g. Clonostachys rosea (Link) Schroers, Samuels, Seifert & W. Gams, Trichoderma 733 viride Pers.) and unidentified mucoraceous and laccase deficient species, but excluding filamentous yeasts.

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735 Table 3. Quasi-binomial GLMM analyses testing effects of location by host species and radial depth on yields 736 of all fungi and basidiomycetes, only (both isolation media combined). 737 Fixed effects Chi square statistic Degrees p-value of freedom All fungi (except yeasts) Location by host species 1.545 3 0.671 Radial depth 8.838 2 <0.05 * Location by species × radial depth 4.550 6 0.602

Confirmed basidiomycetes Location by host species 2.435 3 0.487 Radial depth 2.913 2 0.233 Location by species × radial depth 18.783 6 <0.01 ** 738

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739 Table 4. Mean percentage isolation attempts yielding fungi or just basidiomycetes by location, host

740 (Nothofagus sp.) and radial depth.

741 Type Location Species Radial depth1 0–6 cm 6–12 cm >12 cm All depths No.. % yield No.. % yield No.. % yield No.. % yield isolation isolation isolation isolation attempts attempts attempts attempts All fungi Maruia N. menziesii 784 50 582 47 366 37 1732 46 (except Valley yeasts) Klondyke N. solandri 800 55 576 55 90 28 1466 53 Corner var. cliffortioides Eglinton N. solandri 472 39 285 41 54 41 811 40 Valley var. cliffortioides N. fusca 320 46 240 44 228 39 788 43 Total2 2376 49a 1683 48a 738 37b 4797 47 Confirmed Maruia N. menziesii 784 34 582 31 366 22 1732 30 basidio- Valley mycetes3 Klondyke N. solandri 800 42a 576 41a 90 1b 1466 39 Corner2 var. cliffortioides Eglinton N. solandri 472 14 285 22 54 39 811 19 Valley var. cliffortioides N. fusca 320 27 240 22 228 21 788 23 Total 2376 Draft32 1683 32 738 20 4797 30 742 1No. isolation attempts per sector: 8 (0–6 cm) or 6 (other depth classes); results for both isolation media combined. 743 2Within rows, values with different letters differ significantly (p<0.05, Tukey HSD test). 744 3Excludes a small number of unidentified, non-sporulating isolates producing or deficient in laccase (potentially white and 745 brown rot basidiomycetes, respectively). 746

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747 Table 5. Yields of basidiomycete and ascomycete species (results for both isolation media combined).

Code1 Identity As % of all As % of As % of total No. isolation attempts fungi known per site (in brackets)3 isolated basidio- Maruia Klon- Eglinton Valley (excluding mycetes (N. dyke Boyd East 2 2 yeasts) isolated menz- (N. s . v. (N. s. v. (N. iesii) cliffort- cliffort- fusca) ioides) ioides) (1732) (1466) (811) (788) Ah Armillaria hinnulea Kile & Watling 0.13 0.21 0.17 - - - An Armillaria novae-zelandiae (G. Stev.) 0.80 1.26 - - 1.73 0.51 Herink A Armillaria sp(p). 0.27 0.42 - - 0.74 - At Australoporus tasmanicus (Berk.) 6.91 10.81 7.91 1.09 - 0.25 P.K. Buchanan & Ryvarden B Biscogniauxia sp. (Xylariales) 0.27 - - 0.14 0.49 - b Other unidentified basidiomycetes 4.19 6.56 1.73 1.57 1.48 3.68 Cr1 Crepidotaceae sp. 0.13 0.21 - - 0.37 - Ct Cyclomyces tabacinus (Mont.) Pat. 0.13 0.21 - - - 0.38 D Diatrypella sp. (Xylariales) 0.67 - - - 1.85 - G Ganoderma sp. 11.95 18.69 9.76 - 5.30 7.11 Ga Gloeohypochnicium analogum 0.05 0.07 - 0.07 - - (Bourdot & Galzin) Hjortstam Gy Gymnopus sp.? 0.45 0.70 0.40 - - 0.38 Ho Hohenbuehelia sp. 0.13 0.21 - - 0.37 - H2 Unidentified hymenochaetaceous sp. Draft2.19 3.42 2.83 - - - (associated with fruitbodies of Phellinus kamahi (G. Cunn.) P.K. Buchanan & Ryvarden) H4 Unidentified hymenochaetaceous sp. 10.61 16.60 - 16.24 - - H5 Unidentified hymenochaetaceous sp. 0.67 1.05 - 1.02 - - H6 Unidentified hymenochaetaceous sp. 0.09 0.14 - - - 0.25 In Inonotus nothofagi G. Cunn. 6.73 10.53 6.64 - 4.32 0.13 Mu Mucronella sp. 0.05 0.07 - - - 0.13 P1 Polyporales 1 sp. 0.22 0.35 - - 0.62 - P2 Polyporales 2 sp. 0.05 0.07 0.06 - - - (Scopuloides hydnoides (Cooke & Massee) Hjortstam & Ryvarden?) P3 Laetiporus squalidus 0.49 0.77 - - - 1.40 P5 Polyporales 5 sp. 0.80 1.26 - - - 2.28 Pm Pholiota multicingulata E. Horak 0.05 0.07 - - - 0.13 Pp Pleurotus purpureo-olivaceus (G. 11.41 17.85 - 17.46 - - Stev.) Segedin, P.K. Buchanan & J.P. Wilkie Ppe Postia pelliculosa (Berk.) Rajchenb. 1.74 2.72 - - - 4.95 Ps Peniophorella sp. 0.05 0.07 - - - 0.13 Rc Rigidoporus concrescens (Mont.) 0.54 0.84 0.69 - - - Rajchenb. Rca Ryvardenia campyla (Berk.) 0.22 0.35 - - - 0.64 Rajchenb. S Sistotrema spp. 2.63 4.11 0.12 1.43 3.33 1.14 Sr Schizopora radula (Pers.) Hallenb. 0.27 0.42 - 0.41 - - Xy Xylariaceae sp. (Xylariales) 0.98 - - 1.36 0.25 - ⊕ Laccase positive isolates of 3.97 - 2.95 0.75 2.47 0.89 unidentified white rot species - Other fungi4 30.18 - - - - - Total 100.0 100.0 33.3 41.5 23.3 24.4 Total No. isolates 2243 1434 5765 6095 1895 1925 748 1As adopted in Figs. 3-5. Shading highlights the five most common species. 749 2Numbers of isolates of each species as percentages of total isolates. 750 3Dashes indicate species not isolated (zero yields).

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751 4Comprising hyphomycetes and unidentified mucoraceous and laccase deficient species (⊖; potentially may include 752 some unidentified ascomycetes and brown rot causing basidiomycetes), but excluding filamentous yeasts. 753 5Excludes “other fungi”. 754

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755 Table 6. Species fruiting or showing other signs on study stems. 756 757 Code1 Identity Location/site2 Stem No. NZFRI-M3 A(d) Armillaria sp. (wood decay)4 M 2 - A (m) Armillaria sp. (mycelial fans beneath bark)4 B 8 - A (r) Armillaria sp. (rhizomorphs beneath bark) M 1,2 - At Australoporus tasmanicus K 14,15 - Bc Biscogniauxia capnodes var. capnodes (Berk.) Y.M. Ju M,B 1,6,7 5841 & J.D. Rogers Br Biscogniauxia capnodes var. rumpens (Cooke) Y.M. Ju M,K 5,12,155 5843, 5845 & J.D. Rogers C (s) Chlorociboria sp. (green wood stain) 4 K 146 - Ct Cyclomyces tabacinus E 10 - Fh Fomitopsis hemitephra (Berk.) G. Cunn. M,E 1,97 5844 Ga Ganoderma applanatum sensu Wakefield M,B 1,4,6 5839, 5842 G Ganoderma sp.4 M 4 - Hol Hohenbuehelia luteohinnulea (G. Stev.) E. Horak K 12 5846 Pk Phellinus kamahi M 3 - p Pleurotoid sp.4 B 7 - Sh Stereum hirsutum (Willd.) Pers. M 1 5840 Tv Trametes versicolor (L.) Lloyd K 12,15 - 758 1As adopted in Figs. 3–5. 759 2M, Maruia; K, Klondyke; B, Boyd Creek, Eglinton; E, East Branch, Eglinton. 760 3New Zealand Forest Research Institute Mycology Herbarium, Rotorua, New Zealand. 761 4Not collected for laboratory examination. 762 5Fruiting along stem (not shown in Fig. 5). 763 6At break point. Draft 764 7Side limb from main stem.

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765 Table 7. Occurrence of species found in more than one tree by location and host.1

766 Species Site / tree species / tree identity number code2 Maruia Valley / N. menziesii Eglinton Valley Klondyke Corner / Boyd Ck. / East N. solandri var. cliffortioides N. solandri var. Branch / cliffortioides N. fusca 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 An + + At + + + + + + Bc f f f + Br f f f Cr1 + + Ct + f Fh f f G + + + + + + Gy + + H2 + + + H4 + + In + + + Pp + + + Ppe + + Tv Draft f f Xy + + 767 1By isolation (+) or, where not also isolated, by fruiting (f). Species found in only one tree comprise (tree identity number 768 in brackets): Ah (2), D (7), Ga (12), Ho (7), H5 (11), H6 (9), Hol (f;12), Mu (10), P1 (6), P2 (3), P3 (9), P5 (10), Pk (f;3), 769 Pm (10), Ps (10), Rc (3), Rca (9), Sh (f;1), Sr (12), b14 (4), b27’ (10), b37 (2) and ⊖12 (12). Also excludes fungi not 770 clearly defined as one species (e.g. A, S, b). 771 2As adopted in Tables 5, 6 and Figs. 3–5.

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773 Table 8. Mean richness and diversity indices of basidiomycete fungi in 54 fallen stems of six indigenous 774 species standardised to approximately 120 isolation attempts per stem1. 775 776 Species No. 0D 1D 2D stems (species (Shannon (Simpson richness) diversity) diversity) N. fusca 8 6.38 a 3.87 a 2.98 a N. menziesii 11 5.27 a 3.29 a 2.76 a D. cupressinum 13 3.69 ab 2.48 ab 2.10 ab N. solandri var. 8 3.13 ab 2.26 ab 1.94 ab cliffortioides P. taxifolia 8 2.00 b 1.64 b 1.53 b B. tawa 6 2.33 b 1.54 b 1.36 b Total 54 777 1Higher values indicate greater richness or diversity; within columns, values with the same letter do not differ significantly 778 (p<0.05, Tukey HSD test). 779 780 781 782

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783 784 785

786 787 788 Figure 1.

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Sector 1 a

Sector 4 Sector 2

Ground level Sector 3 789 790 791 Draft 792 793 794 795 796 0 cm 797 798 6 cm 799 12 cm 800 801 802 803 804 b 805 806 807 Figure 2. 808 809 810 811 812 813 814 815 816

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818 819 820 821 822 Figure 3.

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824 825 826 827 828 Figure 4.

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830 831 832 833 834 Figure 5.

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835

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836 Appendix A. Identification of isolates to species, precisely or within a higher taxonomic rank. 837 838 Table A1. Representative isolates identified morphologically &/or by sequencing &/or chemically. Morphology: 839 cultures macro- and micro-morphologically mutually identical and, where noted (fb), matching isolate(s) from 840 identified fruitbodies. Molecular: DNA sequences (with No. base pairs, bp) from morphologically 841 representative isolates show a given % ITS equivalence (i) to selected sequences in GenBank (accession 842 No. in brackets; with % query cover where identified to species), (ii) to sequences of isolates from 843 morphologically identified fruitbodies, or (iii) to sequences of morphologically identical isolates of the same 844 unidentified species from one or more stems, confirming their mutual identity. Chemical: reaction to the stain 845 α naphthol indicating presence (white rot species) or absence (potential brown rot species) of the enzyme 846 laccase. 847 848 Code1 Identity Representative isolates Basis for identification of isolates NZFS2 GenBank sequence Ah Armillaria hinnulea 4536 MH410002 Morphology & molecular (798 bp) (i: EU734747 (NZFS 2942), 99% (94% cover); AF394918, 98% (98% cover); FJ711636, 98% (99% cover)). An Armillaria novae-zelandiae 4541 MH409995 Morphology & molecular (673,813 bp) (i: 4542 MH410001 KR063266 (NZFRI 5645M), 97%,99% (96%,87% cover); FJ660941, 97%,99% (90%,100% cover)). A Armillaria sp(p). - - Morphology (fb). At Australoporus tasmanicus 4504 MH410012 Morphology (fb) & molecular 4506DraftMH410011 (629,644,628,596,529 bp) (ii: NZFS 226, 4559 MH410010 MH409961, ex fb, 98.1–98.7%) (iii: 99.4– 4505 MH410009 100% mutual match between 5 representative 4502 MH410008 isolates from 3 stems). Ref.: Hood et al. 2008. B Biscogniauxia sp. 4507 MH410020 Morphology (isolates resemble those from fb (Xylariales) 4509 MH410021 of B. capnodes) & molecular (782,736,752 bp) 4508 MH410022 (i: JN225898 (ICMP 18828) 3, 100%,100%,97% (97%,99%,98% cover); KU743945, 97%,97%,97% (90%,95%,93% cover)) (iii: 97–100% mutual match between a representative isolate from each of 3 stems). b4 Unidentified - - Morphology (e.g. presence of clamps). basidiomycetes Cr1 Crepidotaceae sp. 4558 MH409983 Morphology & molecular (i) (703,737 bp) 4557 MH409982 (AB509903, 89–90%; HQ728537, 76–78%; KU762016, 88–89%: Crepidotus spp.) (iii: 99.9% match between a representative isolate from each of 2 stems). Ct Cyclomyces tabacinus 4503 MH409976 Morphology (fb) & molecular (608 bp) (i: JQ279517, 98% (99% cover)) (ii: NZFS 2764 ex decayed wood supporting numerous fbs of this species, 98%). Ref.: Hood et al. 2008. D Diatrypella sp. (Xylariales) 4545 MH409964 Morphology, molecular (542 bp) (i: AJ302440, GU062295, LC163518, all 96%; Diatrypella spp.) & chemical (laccase deficient). G Ganoderma sp. 4540 - Morphology (fb). Ref.: Hood et al. 2008. 4538 4539 Ga Gloeohypochnicium 4549 MH409974 Molecular (628 bp) (i: GQ411521 (PDD analogum 91626), 99% (95% cover)). Ref.: Paulus et al. (2007). Gy Gymnopus sp.? 4511 MH409986 Molecular (741,726,533 bp) (i: KY026744, 4521 MH409963 91–92%; KY026619, 91–92%; Gymnopus 4522 MH409987 spp.) (ii: NZFS 2785, ex Marasmius otagensis G. Stev. fb, 89.8 – 93.6%) (iii: 94.3,99.6–

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99.8% mutual match between 4 representative isolates from 2 stems). Ho Hohenbuehelia sp.? 4543 MH409981 Morphology & molecular (684,658 bp) (i: JQ247341, 95%; KU355322, 96%; KX064455, 96%; Hohenbuehelia spp., Nematoctonus sp.) (iii: 99.5% mutual match between 2 representative isolates from opposite ends of 1 stem). H2 Unidentified 4484 MH410005 Morphology & molecular (661,656,663, hymenochaetaceous sp. 4483 MH410004 571,618 bp) (i: KJ668543, 88–89%; (associated with fruitbodies 4490 MH410003 KJ677114, 87–88%; AY558613, 87–89%: of Phellinus kamahi, stem 4491 MH409993 Fuscoporia spp.) (iii: > 99% mutual match 3) 4492 MH409994 between 5 representative isolates from 3 stems). H4 Unidentified 4494 MH409998 Morphology & molecular (539,528,538 bp) (i: hymenochaetaceous sp. 4493 MH409965 KJ206286, 86%; AY072029, 87%; EU035311, 4496 MH409996 87%: Fulvifomes, Inocutis, Phellinus spp.) (iii: 97–99% mutual match between 3 representative isolates from 2 stems). H5 Unidentified 4495 MH409973 Morphology & molecular (747 bp) (i: hymenochaetaceous sp. KC456243; KX181319; AF534073; KT223640; all 90%: Inonotus, Fomitiporella, Phellinus, Tropicoporus spp.) (ii: NZFS 2764, only 56%, see Cyclomyces tabacinus). H6 Unidentified 4498 MH410014 Morphology & molecular (605 bp) (i: hymenochaetaceous sp. AY340053; GQ383780; KU139147; all 91%: Draft Phellinus spp.) In Inonotus nothofagi 4485 MH410007 Morphology & molecular 4497 MH410006 (778,756,156,798,455 bp) (i: GU222327 (PDD 4499 MH409980 98353, I. nothofagi), 99%,99%,99%,97%,99% 4500 MH409979 (92%,95%,100%,90%, 99% cover) (iii: 95– 99.9% mutual match between 4 representative isolates from 2 stems). Mu Mucronella sp.? 4547 MH409972 Molecular (557 bp) (i: KY462452, 98%; HQ533013 (PDD 95742), 86%; EU770252 (ICMP 16979), 84%; Mucronella spp.). P1 Polyporales 1 sp. 4567 MH409984 Morphology & molecular (647 bp) (i: HM583818 (LF219: Eucalyptus obliqua, Tasmania; cf. KF638526), 99%; KY462527, 98%; JN675338, 96%; Ceriporiopsis sp., Phlebia sp.). Ref.: Ortiz et al. (2014). P2 Polyporales 2 sp. 4568 MH410015 Morphology (628 bp) (double clamps present) (Scopuloides hydnoides?) & molecular (i: GU062195, 98%; GU934592, 97%; KP135346, 97%; KP814169, 97%; Phanerochaete sp., Scopuloides sp.) P3 Polyporales 3 sp. 4501 MH409977 Morphology & molecular (461 bp) (i: (Laetiporus squalidus) KP765238, 98%, Laetiporus squalidus; EU840582, 89%; EU840572, 89%; HM583816 (8 AJMH-2010), 96%; HQ332384, 75%; Laetiporus spp.) (ii: NZFS 235, ex Laetiporus portentosus fb, 63%). P5 Polyporales 5 sp. - - Morphology & molecular (656,651 bp) (i: HM583815 (7 AJMH-2010: Eucalyptus obliqua, Tasmania), 99% (87%,88% cover) (iii: 99.9% mutual match between 2 representative isolates from 1 stem) Pm Pholiota multicingulata 4556 MH409971 Molecular (655 bp) (i: HQ832449, 99% (100% cover); HQ533029 (PDD 95841), 99% (100% cover)). Pp Pleurotus purpureo- 4515 MH410000 Morphology & molecular (661,657,598 bp) (i: olivaceus 4512 MH409999 HQ533042 (PDD 95773), 99%,99%,99% 4524 MH409969 (100%,100%,98% cover); GQ411512 (ICMP

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17077), 99%,99%,100% (96%,96%,93% cover); GQ411523 (PDD 91632), 99%,99%,99% (96%,96%,93% cover)) (iii: 99% match between a representative isolate from each of 2 stems). Ref.: Fukami et al. (2010; supporting information). Ppe Postia pelliculosa 4572 MH260263 Morphology (fb) & molecular (679,699 bp) (i: 4573 MH409962 JX090104, 99%,99% (83%,84% cover); JX090103, 98%98% (83%,84% cover); HM583848 (5 AJMH-2010: Eucalyptus obliqua, Tasmania), 98%,98% (91%,92% cover)) (ii: NZFS 2976, MH410017, ex fb, 99.6%, 99.7%; NZFS 2979, MH410018, ex fb, 99.6%, 99.7%). Ref.: Pildain and Rajchenberg (2013). Ps Peniophorella sp. 4548 MH409970 Molecular (691 bp) (i: KP768313, DQ647468, AB907584, all 97% match to P. praetermissa) Rc Rigidoporus concrescens 4516 MH409997 Morphology (fb). Ref.: Hood et al. (2008). Rca Ryvardenia campyla 4520 MH409989 Morphology (fb) & molecular (598,563,340 bp) 4560 MH409988 (i: JQ390051 (NZFS 2826, MH410023), 4574 MH409960 100%,100%,100% (96%,99%,100% cover); JX090118, 100%,100%,100% (94%,99%,100% cover); HM583811, (3 AJMH-2010: Eucalyptus obliqua, Tasmania), 99%,99%,99% (86%,88%,100% cover)) (iii: 99.9–100% mutual match between 3 representative isolates from 1 stem). Refs.: Draft Hood et al. (2008); Hopkins et al. (2011); Pildain and Rajchenberg (2013). S Sistotrema sp(p). 4518 MH409991 Morphology (fb5) & molecular 4519 MH409990 (631,683,651,578,587 bp) (i: respectively: 4523 MH409978 JX507710, 100%; GQ411514 (ICMP 17079), 4534 MH409967 99%; KJ714008, 100%; AY672924, 92%, 4535 MH409966 99%; KM232472, 100%; Sistotrema spp.) (iii: 88–98% mutual match between 5 representative isolates from 3 stems). Ref.: Fukami et al. (2010; supporting information). Sr Schizopora radula 4546 MH409968 Morphology & molecular (599 bp) (i: GQ411525 (PDD 91616), AF145576 (ICMP 13840), AF145566, KT203307, all 99% (98%,90%,90%,100% cover)). Ref.: Paulus et al. (2000). Xy Xylariaceae sp. (Xylariales) 4550 MH410016 Morphology & molecular (602,478 bp) (i: JN225896 (ICMP 18796)6, 99%; JN225895 (ICMP 18770), 98%; JN225894 (ICMP 18791), 98%)(iii: 100% mutual match between a representative isolate from each of 2 stems). Ref.: Johnston et al. (2012). ⊕ Laccase positive isolates of - - Morphology (e.g. lacking clamps) & chemical. unidentified white rot species ⊖4 Laccase deficient isolates - - Morphology (e.g. lacking clamps) & chemical. of unidentified species 849 1As adopted in Table 5 and Figs. 3–5. 850 2NZFS: New Zealand Forest Research Institute Culture Collection, Rotorua, New Zealand. NZFRI-M: New Zealand 851 Forest Research Institute Mycology Herbarium, Rotorua, New Zealand. PDD: New Zealand Fungal Herbarium, Landcare 852 Research, Auckland, New Zealand. ICMP: the International Collection of Micro-organisms from Plants, Landcare 853 Research, Auckland, New Zealand. 854 3Biscogniauxia sp. 1 (ICMP 18828) of Johnston et al. (2012); by contrast, sequences of fruitbody isolates (NZFS 4510, 855 GenBank MH410013; NZFS 4537, GenBank MH410019) of both varieties of B. capnodes from each of two study stems 856 (1, 5) matched (99%,99% (97%,99% cover)) those of Biscogniauxia sp. 2 (ICMP 18793, GenBank JN225897) of 857 Johnston et al. (2012; the wood decay isolates of sp. 1 were from stems 6, 7, and 12). Biscogniauxia capnodes var.

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858 capnodes and B. capnodes var. rumpens, which differ mainly in their ascospore size, showed 99–100% mutual sequence 859 identity using ITS-1F and ITS-4 primers. 860 4An accompanying number in Figs. 3–5 (e.g. b14, ⊖12) denotes a particular, though unidentified species; includes b14 861 (NZFS 4551, GenBank MH409985), b27' (NZFS 4544, GenBank MH409975) and b37 (NZFS 4517, GenBank 862 MH409992). ⊖12 may be Biscogniauxia sp., but not confirmed. 863 5Morphologically matching reference isolates that additionally produced Sistotrema spp. basidia (> 4 sterigmata) in 864 culture. 865 6Xylariaceae sp. 4 (ICMP 18796) of Johnston et al. (2012). 866 867 References 868 869 Fukami, T., Dickie I.A., Wilkie, J.P., Paulus, B.C., Park, D., Roberts, A., Buchanan, P.K., and Allen, R.B. 870 2010. Assembly history dictates ecosystem functioning: evidence from wood decomposer communities. Ecol. 871 Lett. 13: 675–684. doi: 10.1111/j.1461-0248.2010.01465.x _ 2010. 872 873 Hood, I.A., Beets, P.N., Gardner, J.F., Kimberley, M.O., Power, M.W.P., and Ramsfield, T.D. 2008. 874 Basidiomycete decay fungi within stems of Nothofagus windfalls in a Southern Hemisphere beech forest. 875 Can. J. For. Res. 38: 1897–1910. doi:10.1139/X08-041. 876 877 Hopkins, A.J.M., Glen, M., Grove, S.J., Wardlaw, T.J., and Mohammed, C.L. 2011. Wood-inhabiting fungi 878 found within living Eucalyptus obliqua trees in southern Tasmania. Australasian Mycologist 29: 37–46. 879 880 Johnston, P.R., Johansen, R.B., Williams, A.F.R., Wilkie, J.P., and Park, D. 2012. Patterns of fungal diversity 881 in New Zealand Nothofagus forests. Fungal Biol. 116: 401–412. 882 883 Ortiz, R., Párraga, M., Navarrete, J., Carrasco,Draft I., Vega, E. de la, Ortiz, M., Herrera, P., Jurgens, J.A., Held, 884 B.W., and Blanchette, R.A. 2014. Investigations of biodeterioration by fungi in historic wooden churches of 885 Chiloé, Chile. Microb. Ecol. 67: 568–575. doi: 10.1007/s00248-013-0358-1. 886 887 Paulus, B., Hallenberg, N., Buchanan, P.K., and Chamber, G.K. 2000. A phylogenetic study of the genus 888 Schizopora (Basidiomycota) based on ITS DNA sequences. Mycol. Res. 104: 1155–1163. 889 890 Paulus, B., Nilsson, H., and Hallenberg, N. 2007. Phylogenetic studies in Hypochnicium (Basidiomycota), 891 with special emphasis on species from New Zealand. N. Z. J. Bot. 45: 139–150. 892 893 Pildain, M.B., and Rajchenberg, M. 2013. The phylogenetic position of Postia s.l. (Polyporales, 894 Basidiomycota) from Patagonia, Argentina. Mycologia 105: 357–367. doi: org/10.3852/12-088. 895 896 897 898 899

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