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ABSTRACT
ABSTRACT
Cortinarius is a taxonomically diverse genus of ectomycorrhizal fungi. It has a global distribution and is the dominant genus found under Nothofagus in New Zealand. Here we describe Cortinarius atropileatus, associated with Nothofagus. The new species is morphologically very similar to C. dulciolens but may be distinguished by the vivid mauve-violet lamellae when young, spores with reduced verrucose ornamentation and a less pronounced plage, and the presence of abundant clamp connections in the epicuticular hyphae. A molecular phylogenetic analysis of the internal transcribed spacer (ITS) region and RNA polymerase second largest subunit (RPB2) shows that C. atropileatus is in C. sect. Dulciolentes ined. and is sister to an undescribed sequestrate species.
Introduction
The increasing use of DNA metabarcoding to ask ecological questions allows local (Orlovich et al. 2013) and global systems to be described (Tedersoo et al. 2014) with unprecedented scope. However, complex ecological hypotheses that involve the life history of organisms rely on linking DNA barcodes with descriptive taxonomy. For this reason, describing new species, including cryptic ones, remains a critical exercise. Cortinarius (Pers.) Grey is a widespread ectomycorrhizal genus. It is one of the most diverse genera in the world, containing over 2000 species (Matheny et al. 2006). In New Zealand, Cortinarius is associated with Nothofagus, Leptospermum and Kunzea, and it is the most speciose ectomycorrhizal genus associated with Nothofagus (McKenzie et al. 2000). Here we describe a new species of Cortinarius that shares features of agarics previously placed in Cyphocybe R. Heim.
Cuphocybe was erected by Heim (1951) with the type species Cuphocybe olivacea R. Heim (≡ Cortinarius elaiochrous E. Horak, M.M. Moser, Peintner & Vilgalys) and also included Cuphocybe alborosea R. Heim (≡ Cortinarius alboroseus (R. Heim) Peintner, E. Horak, M.M. Moser & Vilgalys), both endemic to New Zealand. Cuphocybe is ectomycorrhizal and shared rough, ornamented spores with Cortinarius, but was delimited from Cortinarius based on the lack of a conspicuous cortina, rather forming pigmented velar fibrils along the length of the stipe (Heim 1951; Horak 1973; Soop 1998). Horak (1973) added Cuphocybe phaeomyxa E. Horak (≡ Cortinarius phaeomyxa (E. Horak) Peintner, E. Horak, M.M. Moser & Vilgalys), associated with Nothofagus fusca and N. menziesii, to the list of New Zealand species. Cuphocybe calospora M. Zang (≡ Cortinarius calosporus (M. Zang) Peintner, M.M. Moser, E. Horak & Vilgalys), associated with Quercus rehderiana forest was described from the eastern Himalayas (Zang 1987) and Cuphocybe ferruginea E. Horak (≡ Cortinarius badioferrugineus E. Horak, Peintner, M.M. Moser & Vilgalys) was described from Papua New Guinea (associated with Castanopsis/Lithocarpus) and New Caledonia (associated with Nothofagus spp.) (Horak 1980). Another New Zealand species, Cuphocybe melliolens Soop (≡ Cortinarius dulciolens E. Horak, M.M. Moser, Peintner & Vilgalys) was described by Soop (1998) from Nothofagus forest.
Using phylogenetic evidence from the analysis of internal transcribed spacer (ITS) region and large ribosomal subunit (LSU) markers, Peintner et al. (2002a) showed that Cuphocybe is not monophyletic, and that the three New Zealand species were all nested within Cortinarius. Thus, the grouping of Cuphocybe-like species on morphological characters is artificial. All Cuphocybe species were subsequently transferred to Cortinarius (Peintner et al. 2002b).
Cortinarius dulciolens is a sweet-smelling agaricoid fungus associated with southern beech (Nothofagus spp.) in New Zealand. It is closely related to the sequestrate C. beeverorum Orlovich, X. Yue Wang & T. Lebel (Orlovich et al. 2014), and both species are in C. sect. Dulciolentes ined. (Soop 2017). Here we describe another new species in C. sect. Dulciolentes, Cortinarius atropileatus A. R. Nilsen & Orlovich from New Zealand associated with southern beech (Nothofagus) species.
Materials and methods
Morphological examination
Eleven collections of C. dulciolens (OTA62181, OTA62338, OTA62375, OTA62399, OTA62401, OTA62406, OTA64053, OTA64056, OTA65696, OTA69682, OTA70444) and six collections of C. atropileatus (OTA60170, OTA70337, OTA70420, OTA70421, OTA70423, OTA70424) were compared for morphological analysis. Material for examination was hand-sectioned and mounted in 5% (w/v) KOH. Measurements were made using a compound light microscope (Olympus CH2) under oil immersion. Spore measurements are stated as the mean ±1 SD of 30 measurements from each collection examined, with minimum and maximum values reported in parentheses. Spore measurements exclude the apiculus and ornamentation. Q value (ratio of length to width) is reported as the mean ±1 SD. Basidium measurements are reported as the mean length and width of 20 measurements, with minimum and maximum values reported in parentheses. Differences in spore length and width between species were tested by a t-test in base R (R Core Team 2017). Morphological terminology follows Largent (1986) and Largent et al. (1977). Chemical reactions with KOH 5% (w/v) were performed on dried herbarium specimens.
Scanning electron microscopy of spores was achieved by gently pressing and removing dried lamellae fragments on to carbon tape mounted on stubs. Specimens were coated with gold/palladium to a thickness of 10 nm using a sputter coater (Emitech K575X). Electron micrographs of spores were taken with a scanning electron microscope (JEOL 6700F FE-SEM). Scanning electron micrographs were used to aid the description of spore ornamentation, however spore descriptions are based on observations made under a light microscope, unless otherwise stated.
DNA extraction, PCR and sequencing
DNA was extracted from dried herbarium specimens using a Plant Genomic DNA Mini Kit (Geneaid) following the manufacturer’s protocol. Polymerase chain reactions (PCR) of the internal transcribed spacer (ITS) region and RNA polymerase II (RPB2) gene were performed in 50 µL reactions using Platinum Taq (Invitrogen). Each reaction contained 1× buffer, 1.5 mM MgCl2, 0.2 mM dNTP mix, 10 pmol ITS1f (Gardes and Bruns 1993) and ITS4 (White et al. 1990) for the ITS region, or 10 pmol bRPB2-6F and bRPB2-7.1R (Matheny 2005) for the RPB2 gene, 1 µL 1:10 diluted DNA in double distilled H2O and 2 U Platinum Taq. The PCR protocol for ITS was 94°C for 2 min, then 35 cycles of 94°C for 30 s, 55°C for 30 s, 72°C for 1 min, then 72°C for 5 min. The PCR protocol for the RPB2 gene was 94°C for 2 min, then 35 cycles of 94°C for 30 s, 50°C for 30 s, increase 0.3°C/s to 72°C, 72°C for 45 s, then 72°C for 5 min. PCR products were purified using a PureLink PCR Purification Kit (Invitrogen) following the manufacturer’s instructions, quantified using a spectrophotometer (Nanodrop ND-2000c) and sequenced using BigDye Terminator v.3.1 cycle sequencing (Life Technologies) by Genetic Analysis Services, Department of Anatomy, University of Otago.
Alignment and phylogenetic analysis
Electropherograms were assembled and edited using Geneious 10.1.3 (Biomatters). Additional sequences that had high blast matches to C. atropileatus were retrieved from GenBank. Cortinarius porphyroideus (PDD103546) was selected as the outgroup. Sequences were aligned using the MUSCLE plugin in Geneious R10.1.3, with a maximum of 16 iterations. Obviously misaligned bases were manually corrected. GenBank accession numbers of all sequences used in this study are indicated in . The alignment is available on TreeBASE Study Accession URL: http://purl.org/phylo/treebase/phylows/study/TB2:S23363. Preliminary phylogenetic analyses indicated that the ITS and RPB2 phylogenies were compatible, and thus the alignments were combined. The resulting concatenated alignment was 1574 characters in length.
Table 1. Species and sequences used in this study.
The alignment was analysed by maximum parsimony analysis using PAUP* 4.0b10 for Unix (Swofford 2003). Of the 1574 characters, 302 were parsimony informative and 1272 were excluded from the analysis. Most parsimonious trees were found by 1000 replicate heuristic searches with the tree bisection-reconnection algorithm, saving 100 trees per replicate and a maximum of 10,000 trees. Branch support was determined by bootstrap analysis of 1000 replicates, with each replicate comprising 5 replicate heuristic searches, saving 10 trees per replicate. Results of the bootstrap analysis were transferred to corresponding branches on the Bayesian inference tree.
For the Bayesian inference analysis, we found that analyses based on partitioning ITS into separate gene/spacer partitions (18S, ITS1, 5.8S, ITS2 and 28S) and partitioning RPB2 into 1st, 2nd and 3rd codon positions, were topologically identical to a tree from an analysis with only 2 partitions, i.e. ITS and RPB2. Therefore, the alignment was partitioned into two partitions only, based on the separate marker regions. The ‘greedy’ search scheme was used to determine the best DNA substitution models in Partitionfinder2 (Guindon et al. 2010; Lanfear et al. 2017). The alignment was analysed by Bayesian inference using MrBayes 3.2.6 (Ronquist and Huelsenbeck 2003), with the following models applied to the partitions: ITS: HKY + G + I; RPB2: SYM + G. Four independent Markov chains were run for 1 × 107 generations, sampling every 100 generations. The runs were determined to have converged when the average standard deviation of split frequencies was less than 0.01. A Bayesian inference 50% majority rule tree and posterior probability values were estimated from the samples after discarding the first 25% (25,000) of sampled trees.
Trees from both analyses were rooted on the branch leading to the outgroup species Cortinarius porphyroideus (PDD103546). Phylogenetic trees were visualised in FigTree v1.4.3 (http://tree.bio.ed.ac.uk/software/figtree/).
Results
Comparison of morphological characters of C. atropileatus to C. dulciolens
The characters examined in C. dulciolens collections were in accordance with the descriptions in Soop (1998, 2017). Both C. dulciolens and C. atropileatus have a brown to black pileus and a pale stipe, and are difficult to separate macroscopically. Spores of C. atropileatus had a shorter mean length (12.1 µm, n = 180) and a narrower mean width (7.0 µm, n = 180) than C. dulciolens (mean length: 13.2 µm (n = 330), mean width: 7.4 µm (n = 330), p < 0.0001) (, Figures 1C & 2B). However, both species have an overlapping range of spore sizes. Comparison of scanning electron micrographs of approximately 10 spores made from two collections of each species indicated that there is minimal variation between species. Representative images from the type specimen of C. atropileatus (Figure 3A–C) and C. dulciolens (Figure 3D–F) illustrate the minute differences in spore morphology. Cortinarius atropileatus had a less distinct plage and small nodules connected by fine ridges that had a web-like appearance. In contrast, the nodules in C. dulciolens were larger and often formed plateaus that were connected by robust ridges. These subtle differences are not apparent in the light microscope (Figures 1C and 2B).
Published online:
27 November 2018Figure 1. Cortinarius atropileatus holotype (OTA70424): A, Basidiocarps, scale = 1 cm; B, Pellis, illustrating the epicutis (EPI), hypocutis (HYP) and cortex (C), scale = 25 µm; C, Basidiospores, scale = 10 µm.
![]({"type":"image","src":"/na101/home/literatum/publisher/tandf/journals/content/tnzb20/2019/tnzb20.v057.i01/0028825x.2018.1548493/20190227/images/medium/tnzb_a_1548493_f0001_oc.jpg"})
Figure 1. Cortinarius atropileatus holotype (OTA70424): A, Basidiocarps, scale = 1 cm; B, Pellis, illustrating the epicutis (EPI), hypocutis (HYP) and cortex (C), scale = 25 µm; C, Basidiospores, scale = 10 µm.
Published online:
27 November 2018Figure 2. Cortinarius dulciolens (OTA62181): A, Pellis, illustrating the epicutis (EPI), hypocutis (HYP) and cortex (C), scale = 25 µm; B, Basidiospores, scale = 10 µm.
![]({"type":"image","src":"/na101/home/literatum/publisher/tandf/journals/content/tnzb20/2019/tnzb20.v057.i01/0028825x.2018.1548493/20190227/images/medium/tnzb_a_1548493_f0002_oc.jpg"})
Figure 2. Cortinarius dulciolens (OTA62181): A, Pellis, illustrating the epicutis (EPI), hypocutis (HYP) and cortex (C), scale = 25 µm; B, Basidiospores, scale = 10 µm.
Published online:
27 November 2018Figure 3. Scanning electron micrographs of spores: A–C, C. atropileatus (OTA70424); D–F, C. dulciolens (OTA62181). Scale = 1 µm.
![]({"type":"image","src":"/na101/home/literatum/publisher/tandf/journals/content/tnzb20/2019/tnzb20.v057.i01/0028825x.2018.1548493/20190227/images/medium/tnzb_a_1548493_f0003_ob.jpg"})
Figure 3. Scanning electron micrographs of spores: A–C, C. atropileatus (OTA70424); D–F, C. dulciolens (OTA62181). Scale = 1 µm.
Table 2. Comparative morphological characters of C. dulciolens and C. atropileatus.
Taxonomy
Cortinarius atropileatus A. R. Nilsen & Orlovich, sp. nov.
(Figure 1 & Figure 3 A–C), MycoBank no.: MB 823338
Type: New Zealand: Catlins, Catlins River Walk, on soil under Nothofagus menziesii, 1 July 2017. Holotype OTA70424.
Diagnosis: C. atropileatus is distinguishable from C. dulciolens by the reduced verrucose spore ornamentation, less pronounced plage, more vivid mauve-violet lamellae when young, more abundant clamp connections in the epicuticular hyphae, and lack of encrustations in the tramal cell walls.
Etymology: atropileatus pertains to the dark-coloured pileus
Description: Pileus convex, 25–55 mm diam., subviscid, dark brown to almost black, innate fibrillose to granulose, margin paler. Lamellae mauve to violet when young becoming brown with age, close, adnexed. Stipe cylindrical to subclavate, occasionally with an adaxial protrusion at base, 40–60 × 8–18 mm, white to pale brown, fibrils from base becoming more granular-dotted towards the apex. Context white becoming pale yellow-brown with age. Odour sweet/honey-like, taste not recorded. Macrochemical reactions KOH dark brown on cutis and gills, weakly red-brown on stipe and stipe cortex. Spores yellow-brown, 12.1 ± 0.9 (9–15) × 7.0 ± 0.5 (6–8) µm, Q = 1.7 ± 0.2, elliptic to subamygdaloid, moderately verrucose, ornamented with irregular nodules, some with faint plage, inamyloid. Basidia hyaline, occasionally stain brown in KOH, 36.5 (30–45) × 12 (11–17) µm, clavate, clamped, sterigmata to 5 μm, 4-spored. Basidioles clamped. Cystidia not observed. Epicutis undulating hyphae with abundant clamps, hyaline to yellow-brown pigmented hyphae, sporadically branching, weak to moderate epimembranous encrustations, hyphae 2.5–4 µm diam. Hypocutis comprised of irregularly shaped cells, 15–30 × 20–60 µm, yellow-brown pigmented some with brown encrustations in cell walls, clamp connections not observed. Context cells weakly to strongly yellow-brown pigmented, clamps present. Trama hyphae hyaline to weak brown without encrustations in the cell walls, 11–30 × 40–80 µm, clamps present.
Habitat: solitary to gregarious on soil, found under Nothofagus spp. and mixed Nothofagus/Leptospermum.
Distribution: South Island, collections are limited to Charleston, West Coast and the Catlins.
Collections examined: New Zealand, South Island: Catlins, Catlins River Walk, on soil under Nothofagus menziesii, 21 April 2017, leg. A. R. Nilsen (OTA70337, OTA70423, OTA70424), 28 May 2017, leg. A. R. Nilsen (OTA70420, OTA70421). Westcoast, Charleston, Tiropahi Creek, on soil under Leptospermum and beech, 11 May 2006, leg. D. A. Orlovich (OTA60170).
Phylogenetic position of C. atropileatus
The maximum parsimony analysis resulted in 5990 equally parsimonious trees of length 848, each with consistency index = 0.529, retention index = 0.790 and a rescaled consistency index = 0.418. The maximum parsimony tree was topologically compatible with the Bayesian inference tree, and the maximum parsimony bootstrap values were transferred to the corresponding branches on the Bayesian tree (Figure 4). After 1 × 107 generations, the standard deviation of split frequencies was 0.001993. The Potential Scale Reduction Factor was 1.000, and the minimum Estimated Sample Size was greater than 3800, for all parameters sampled.
Published online:
27 November 2018Figure 4. Bayesian inference phylogeny of concatenated internal transcribed spacer (ITS) region and RNA polymerase second largest subunit (RPB2) sequences. Posterior probability values are indicated above branches and bootstrap (%) values from the maximum parsimony analysis are indicated below the branches. Sequestrate fungi denoted by solid circles. Clades are named following the classification of Soop (2017).
![]({"type":"image","src":"/na101/home/literatum/publisher/tandf/journals/content/tnzb20/2019/tnzb20.v057.i01/0028825x.2018.1548493/20190227/images/medium/tnzb_a_1548493_f0004_ob.jpg"})
Figure 4. Bayesian inference phylogeny of concatenated internal transcribed spacer (ITS) region and RNA polymerase second largest subunit (RPB2) sequences. Posterior probability values are indicated above branches and bootstrap (%) values from the maximum parsimony analysis are indicated below the branches. Sequestrate fungi denoted by solid circles. Clades are named following the classification of Soop (2017).
Collections of C. atropileatus were monophyletic on the Bayesian tree (Figure 4) and were sister to an undescribed, sequestrate fungus from New Zealand (PDD106018) (PP = 1, BS = 100). The clade containing C. atropileatus is sister to the clade formed by C. beeverorum and C. dulciolens. The relationship between C. dulciolens and C. beeverorum was well supported (PP = 0.99, BS = 78%). Within C. dulciolens, collections fell into two clades in the maximum parsimony analysis, one of those clades was also present in the Bayesian analysis (PP = 1, BS = 97%). The remaining collections of C. dulciolens comprised another clade in the parsimony analysis supported by a bootstrap value of 64%, but those collections, which included the holotype PDD68471, were not monophyletic in the Bayesian analysis. Within C. dulciolens, two genotypes of collections differ at 7 sites in the ITS region and 4 sites in RPB2. We found no differentiating macroscopic or microscopic morphological characters that separated collections within the C. dulciolens clade.
Cortinarius atropileatus is nested in a strongly supported clade (PP = 1, BS = 97%) corresponding to C. sect. Dulciolentes ined. (Soop 2017), which is dominated by sequestrate species. Of the taxa present in this section, only three have agaricoid fruitbodies: C. atropileatus, C. dulciolens and C. corrugatus. Interestingly, C. corrugatus has been observed to have a gasteroid form (R. E. Halling, https://www.nybg.org/bsci/res/hall/corrug.html). The remaining taxa all have a sequestrate habit: Cortinarius sp. (PDD106018, T833, H1213, H5791), C. beeverorum, Cortinarius atratus (MEL2057704, Trappe18831, TUB011923, Massee270) and C. peraurantiacus (PDD70818 and PDD80481).
Discussion
Cortinarius atropileatus is recognisable as a cuphocyboid Cortinarius. Soop (2017) recorded five species in the morphological group Cuphocybe (C. dulciolens, C. alboroseus, C. canovestitus, C. elaiochrous and C. phaeomyxa), based on an absent or rudimentary cortina, cylindrical stipe with a piston-like or rounded bulb, large, elliptic spores and epimembranal encrustations in the epicutis. Cortinarius atropileatus is distinguishable from most other cuphocyboid species: C. alboroseus is pale in colour and has a pink hue to its pileus and stipe, C. canovestitus is similar to C. alboroseus but lacks a pink tinge, C. elaiochrous is a glutinous fungus with olive hues, and C. phaeomyxa has a chocolate-brown pileus with conspicuous ochre patches or squamules. Cortinarius atropileatus closely resembles C. dulciolens macroscopically, but can be distinguished by microscopic characters and is phylogenetically distinct. Similar to C. dulciolens, C. atropileatus exhibits a sweet, honey-like fragrance. Cortinarius sect. Defibulati also possesses a rudimentary or absent cortina but is distinguished from the species previously described in Cuphocybe based on hyphae lacking clamp connections (Soop 2014).
Within C. dulciolens, some genetic variation is evident. Inspection of macroscopic and microscopic characters did not reveal any discernible intraspecific differences corresponding with the two genotypes identified. The lack of morphological differentiation and the subtle genetic variation may be an indication of an early speciation event or a slowdown in morphological character evolution (Egea et al. 2016).
Sister to C. atropileatus is an undescribed sequestrate species (PDD106018) that has a strong morphological affinity to C. porphyroideus, with a purple basidiocarp and secotioid habit. This species is also associated with Nothofagus species. Noteworthy is that C. section Dulciolentes has a high proportion of sequestrate species. Although sequestrate species are not uncommon in Cortinarius, clades that contain multiple sequestrate species are less common. A large meta-analysis of the evolution of Australian sequestrate fungi (Sheedy et al. 2016) showed sequestration within Cortinarius occurred 12 times, with 7 transformations being autapomorphic and five transformations synapomorphic for clades of 2–5 species. Large species radiations from a sequestrate ancestor are determined to have occurred in the genera Turmalinea and Rossbeevera (Orihara et al. 2016).
Cortinarius atropileatus is associated with Nothofagus in New Zealand. While myrtaceous species (Leptospermum and Kunzea) are recorded co-occurring with Nothofagus in some collections studied here, C. atropileatus is not recorded from habitats without Nothofagus present. Similarly, C. dulciolens is recorded from Nothofagus forests in New Zealand and has an overlapping distribution with C. atropileatus. Cortinarius dulciolens collections OTA69682 and OTA70444 were made in the same location as C. atropileatus collections OTA70337, OTA70423, OTA70424, OTA70420 and OTA70421. Due to the overlapping distribution, ecology and morphological characters of C. atropileatus and C. dulciolens, identification of these species in the field is difficult.
| Species | Collection | ITS | RPB2 |
|---|---|---|---|
| Cortinarius achrous | PDD107695 | KT875174 | — |
| Cortinarius alboroseus | PDD105432 | MH101566 | — |
| Cortinarius altissimus | TENN069830 | NR_153032 | — |
| Cortinarius atrotomentosus | TENN065527 | NR_153030 | KJ920081 |
| Cortinarius atropileatus | OTA60170 | JX178605 | MG367643 |
| OTA70337 | MG367630 | MG367644 | |
| OTA70420 | MG367631 | MG367645 | |
| OTA70421 | MH023302 | MH032582 | |
| OTA70423 | MH023301 | — | |
| OTA70424 | MG367632 | MG367646 | |
| Cortinarius beeverorum | OTA60155 | KC520546 | MG367637 |
| Cortinarius canovestitus | PDD103667 | KF727353 | — |
| Cortinarius caperatus | TUB020420 | KJ421125 | — |
| Cortinarius carneipallidus | PDD95444 | NR153028 | KJ920076 |
| Cortinarius cartilagineus | PDD105768 | MH101571 | — |
| Cortinarius corrugatus | IB2000544 | AF325611 | — |
| Cortinarius dulciolens | Horak NZ8635 | AF325610 | — |
| OTA62181 | MG367627 | MG367640 | |
| OTA62338 | KF977697 | MG367647 | |
| OTA62375 | MH023298 | — | |
| OTA62399 | MH023297 | MH032580 | |
| OTA62401 | MH023296 | MH032579 | |
| OTA62406 | MH023300 | MH032581 | |
| OTA64053 | MH023295 | MH032578 | |
| OTA64056 | MG367633 | MG367648 | |
| OTA65696 | MG367629 | MG367642 | |
| OTA69682 | MG367634 | MG367649 | |
| OTA70444 | MH023299 | — | |
| PDD68471 | JX648592 | — | |
| Cortinarius elaiochrous | PDD77478 | AY669627 | — |
| Cortinarius ECM root tip clone | WME21 | JN847466 | — |
| Cortinarius epiphaeus | PDD103876 | KF727356 | — |
| Cortinarius hallowellensis | MEL2350466 | KJ920007 | KJ920098 |
| Cortinarius indolicus | PDD103881 | KF727396 | — |
| Cortinarius kioloensis | PDD99309 | KJ920013 | KJ920070 |
| Cortinarius majestaticus | OTA65727 | KU523929 | KU523960 |
| Cortinarius meleagris | PDD72781 | HM060324 | — |
| Cortinarius naphthalinus | PDD70505 | GU233344 | — |
| Cortinarius ohauensis | PDD103877 | KF727355 | — |
| Cortinarius palatinus | ZT10422 | KJ920027 | KJ920082 |
| Cortinarius peraurantiacus | PDD70818 | KC520543 | — |
| PDD80481 | KC520544 | — | |
| Cortinarius phaeomyxa | PDD107511 | MG019355 | — |
| Cortinarius porphyroideus | PDD103546 | KT334128 | KT334151 |
| Cortinarius pseliocaulis | PDD107696 | KT875176 | — |
| Cortinarius pselioticton | PDD88277 | JX000371 | — |
| Cortinarius similis | HKAS26154 | AY669577 | — |
| Cortinarius sp. | H1213 | DQ328093 | — |
| H5791 | DQ328099 | — | |
| T833 | JF960730 | — | |
| PDD72554 | MH101519 | — | |
| PDD106018 | MH101579 | — | |
| Trappe26352 | JX983155 | — | |
| Cortinarius subcastanellus | PDD95557 | MH101547 | — |
| Cortinarius taylorianus | NZ858 | AY033106 | — |
| Cortinarius trichocarpus | PDD103637 | KF727354 | — |
| Cortinarius violaceus | TUB011825 | AY669579 | — |
| Cortinarius wallacei | PDD97543 | KJ635242 | — |
| Cortinarius atratus [≡ Protoglossum luteum] | Massee270 | EU084983 | — |
| MEL2057704 | DQ328197 | — | |
| Trappe18831 | AF325612 | — | |
| TUB011923 | AY669606 | — |
GenBank accession numbers in bold were generated for this study.
| Cortinarius dulciolens | Cortinarius atropileatus | |
|---|---|---|
| Lamellae | Pale cinnamon to argillaceous, sometimes with a violet tinge, crowded | Mauve to violet when young becoming brown with age, close, adnexed |
| Mean spore length (μm) | 13.3 | 12.1 |
| Mean spore width (μm) | 7.4 | 7.0 |
| Spore ornamentation | strong | strong |
| Plage | strong | weak |
| Epicutis encrustations | weak-moderate | weak-moderate |
| Epicutis clamps | rare | abundant |
| Hypocutis encrustations | brown, moderate-strong | brown, weak-moderate |
| Hypocutis clamps | rare | rare |
| Trama encrustations | brown, moderate-strong | not observed |
| Trama clamps | occasional | occasional |
Acknowledgements
We thank Eric McKenzie, Shaun Pennycook and an anonymous reviewer for their helpful comments, and Jerry Cooper for assistance locating literature. We thank the New Zealand Department of Conservation for permission to collect from national parks and reserves.
Disclosure statement
No potential conflict of interest was reported by the authors.
ORCID
Andy R. Nilsen http://orcid.org/0000-0003-4724-6289
Christopher M. Brown http://orcid.org/0000-0003-0079-7067
Tina C. Summerfield http://orcid.org/0000-0003-1878-7019
David A. Orlovich http://orcid.org/0000-0002-9266-2188