01-Phylogenetic studies in Hypochnicium _Basidiomycota__ with special emphasis on species from Ne

Prévia do material em texto

Full Terms & Conditions of access and use can be found at
https://www.tandfonline.com/action/journalInformation?journalCode=tnzb20
New Zealand Journal of Botany
ISSN: 0028-825X (Print) 1175-8643 (Online) Journal homepage: www.tandfonline.com/journals/tnzb20
Phylogenetic studies in Hypochnicium
(Basidiomycota), with special emphasis on species
from New Zealand
Barbara Paulus, Henrik Nilsson & Nils Hallenberg
To cite this article: Barbara Paulus, Henrik Nilsson & Nils Hallenberg (2007) Phylogenetic
studies in Hypochnicium (Basidiomycota), with special emphasis on species from New Zealand,
New Zealand Journal of Botany, 45:1, 139-150, DOI: 10.1080/00288250709509709
To link to this article: https://doi.org/10.1080/00288250709509709
Published online: 18 Feb 2010.
Submit your article to this journal 
Article views: 522
View related articles 
Citing articles: 1 View citing articles 
https://www.tandfonline.com/action/journalInformation?journalCode=tnzb20
https://www.tandfonline.com/journals/tnzb20?src=pdf
https://www.tandfonline.com/action/showCitFormats?doi=10.1080/00288250709509709
https://doi.org/10.1080/00288250709509709
https://www.tandfonline.com/action/authorSubmission?journalCode=tnzb20&show=instructions&src=pdf
https://www.tandfonline.com/action/authorSubmission?journalCode=tnzb20&show=instructions&src=pdf
https://www.tandfonline.com/doi/mlt/10.1080/00288250709509709?src=pdf
https://www.tandfonline.com/doi/mlt/10.1080/00288250709509709?src=pdf
https://www.tandfonline.com/doi/citedby/10.1080/00288250709509709?src=pdf
https://www.tandfonline.com/doi/citedby/10.1080/00288250709509709?src=pdf
New Zealand Journal of Botany, 2007, Vol. 45: 139-150
0028-825X/07/4501-0139 © The Royal Society of New Zealand 2007
Phylogenetic studies in Hypochnicium (Basidiomycota), with special
emphasis on species from New Zealand
BARBARA PAULUS
Landcare Research
Private Bag 92170
Auckland 1142, New Zealand
HENRIK NILSSON
NILS HALLENBERG
Department of Plant and Environmental Sciences
Carl Skottsberg Gata 22b
S-405 30 Goteborg, Sweden
Abstract The taxonomic and phylogenetic affilia-
tion of Hypochnicium specimens from New Zealand
were studied using morphological and molecular
analyses. Phylogenetic analyses based on ITS re-
gion of rDNA suggest that species within Hypoch-
nicium form a monophyletic group with respect to
outgroups. The synonymy of Gyrophanopsis with
Hypochnicium is validated and the genus Nodotia
is re-evaluated as a synonym of Hypochnicium.
Sequence analyses of H. zealandicum and a speci-
men within the H. punctulatum complex from New
Zealand place both as sister taxa of H. polonense
and H. cremicolor, respectively. Although spore
dimensions of H. zealandicum overlap with those of
H polonense, it is currently retained as a separate
species on the basis of geographical separation and
phylogenetic analysis. New Zealand specimens re-
corded as H. punctulatum represent a new species,
which is described and illustrated as H aotearoae.
Keywords corticioid fungi; Gyrophanopsis;
Hypochnicium; Nodotia; phylogeny; ITS
B06020; Online publication date 29 March 2007
Received 13 June 2006; accepted 20 December 2006
INTRODUCTION
A considerable part of all fungi are involved in the
decomposition of wood and play key roles in nutrient
cycling and food webs. Among these are corticioid
fungi, a species-rich group of basidiomycetes. They
are characterised by crust-like (resupinate) fruiting
bodies which frequently look like a splash of paint
on the surface of decaying wood. From a systematic
point, this is an unnatural group, distributed among
all major evolutionary lineages in the Homobasidio-
mycetes (Larsson et al. 2004). Corticioid fungi are
an understudied group, particularly in the Southern
Hemisphere, and it is likely that many more species
await discovery. InNew Zealand, a major treatment
of corticioid fungi was by Cunningham (1963). He
described numerous taxa as new to science and re-
corded others that resembled Northern Hemisphere
taxa, using Northern Hemisphere names. New Zea-
land's isolated location and Southern Hemisphere
connections place doubt on the applicability of some
of these names. The advent of molecular meth-
ods now provides the means to determine whether
Northern Hemisphere names should be applied to
New Zealand corticioid fungi. In addition, many
species and generic concepts have changed since
then but only a few of Cunningham's taxa have been
reviewed and updated (e.g., Stalpers 1985; Stalpers
& Buchanan 1991; Parmasto et al. 2004).
Eriksson (1958) introduced Hypochnicium for
corticioid species with distinct and richly branched
hyphae and, more importantly, rather large, thick-
walled spores, and spore walls that stain with cotton
blue (= cyanophilous). While Eriksson & Ryvarden
(1976) considered Hypochnicium a well-circum-
scribed genus, they recognised that som e subgroups
among the North European species were distinct
enough to form separate subgenera. For example, H
polonense (Bres.) Strid is distinguished from other
species by its cylindrical, septate cystidia and distinct
basal hyphae. A similar species was described from
New Zealand under the name Pellicularia zealandica
139"
140 New Zealand Journal of Botany, 2007, Vol. 45
by Cunningham (1953). Stalpers & Buchanan (1991)
revised species oiPeniophora and Pellicularia de-
scribed by Cunningham. They accepted the Julich
(1979) combination Gyrophanopsis zealandica and,
recognising its close relationship with/f. polonense,
formed a new combination, G. polonensis, for that
species also. In his checklist to genera and species of
corticioid fungi, Hjortstam (1997) moved both taxa
to Hypochnicium but did not provide a rationale.
This view was supported by Boidin & Gilles (2000)
who also accepted H lyndoniae as a Hypochnicium
species.
The great variation in some species of Hypoch-
nicium, in characters such as spore dimension and
shape, suggests the presence of species groups.
Recently, Nilsson & Hallenberg (2003) clarified
the phylogeny of the H. punctulatum complex and
recognised four separate species, H albostramineum
(Bres.) Hallenb., H. cremicolor Bres., H punctula-
tum (Cooke) J.Erikss., and H wakefieldiae (Bres.)
J.Erikss. The morphological separation was under-
taken primarily on the basis of spore dimensions,
although overlapping ranges of spore sizes were
recorded for/f. punctulatum and/f. albostramineum.
Two further species have been reported within the
H punctulatum complex. Hypochnicium cystidiatum
Boidin & Gilles was described from the Central
African Republic and has been also reported from
Gabon (Boidin & Gilles 1971) and India (Hj ortstam
&Larsson 1994). From New Zealand, amemberof
the H punctulatum complex was reported as Corti-
cium punctulatum Cooke (Cunningham 1963). Hy-
pochnicium cystidiatum and C. punctulatum sensu
Cunningham have morphological affinities, but their
relationship within the genus has not been clari-
fied.
Another species at one tim e placed in Hypochnici-
um is H lyndoniae (D. AReid) Hjortstam, which was
described as Odontia lyndoniae from Australia (Reid
1956) and was recorded by Cunningham (1959) for
New Zealand. It has been recently transferred to
the genus Nodotia Hjortstam, which was reintro-
duced to accommodate Hypochnicium species with
thick-walled elements ("skeletocystidia") in their
spines or "teeth" (Hjortstam & Ryvarden 2004).
Gloeohypochnicium analogum (Bourdot & Galzin)
Hjortstam was initially included in Hypochnicium
on the basis of its cyanophilous, thick-walled spores
but later transferred to its own genus based on the
presence of gloeocystidia, i.e., cystidia that darken
in sulfo-vanilline (Eriksson & Ryvarden 1976).
This decision was later supported by phylogenetic
analyses (Binder et al. 2005).
The aim of the present study was to examine the
taxonomic position of some New Zealand specimens
of Hypochnicium and to add to our understanding of
the phylogenetic relationships within the genus.
METHODS
The taxa sampled for this study represented a wide
rangeof morphological subgroups and utilised au-
thenticated fungal cultures accumulated in the Fun-
gal Culture collection of the University of Goteborg
(FCUG) during 25 years of research. For compara-
tive purposes, cultures of H cystidiatum were also
acquired from the collection of Boidin, now held at
the Mycotheque de l'Universite catholique de Lou-
vain (MUCL). Both culture collections have been
developed with the purpose to carefully document
species delimitations and, therefore, are highly reli-
able. Additional New Zealand material held at Her-
barium PDD was also examined. For the molecular
analyses, the selected outgroup taxa were the closest
known representatives for which sequences could be
obtained. Outgroup taxa and Hypochnicium belong
to a group of residual taxa identified by Binder et al.
(2005) that, although positioned within the polypo-
roid clade, do not belong to the "core polypores".
Sequence data were generated from the ITS re-
gion for nine specimens in Hypochnicium, and a
further 17 sequences were included from GenBank
(Table 1). Primers and protocols used for DNA iso-
lation, PCR amplification, and sequencing follow
Nilsson & Hallenberg (2003). In brief, mycelia were
harvested from agar and dried between sheets of
sterile filter paper; approximately 2 mg (dry weight)
of input mycelium were used per specimen. DNA
extraction was undertaken using the DNeasy® Plant
Mini Kit (QIAGEN®, Hilden). The instructions of
respective manufacturers were followed for DNA
extraction and all following steps of DNA prepara-
tion, purification, and sequencing. The polymerase
chain reactions were carried out using READY-
TO-GO™ PCR Beads kits, the PCR primers ITS1F
and ITS4B, and the PCR set-up of Gardes & Bruns
(1993). The PCR product was purified using the
QIAqmck™ Spin Procedure (QIAGEN, Hilden).
Sequencing reactions were conducted using 100 ng
of template DNA and the CEQ 2000 Dye Terminator
Cycle Sequencing with Quick Start kit (Beckman
Coulter, Fullerton). Sequences were obtained using
the CEQ 2000XL DNAAnalysis System (Beckman
Coulter) and edited in Sequencher® 4 (GeneCodes
Inc., AnnArbor). Initially, sequences were aligned in
Paulus et al.—Hypochnicium from New Zealand 141
Table 1 Specimens and cultures used in the phylogenetic analysis and morphological examination. Specimens
were utilised in phylogenetic analysis (P) and/or morphological examination (M). FCUG refers to the Fungal Culture
Collection of the University of Goteborg. Herbarium acronyms follow Holmgren & Holmgren (1998).
Taxon FCUG no. Collection no.
Hypochnicium albostramineum (Bres.) Hallenb.
P,M
P,M
H aotearoae sp. nov.
M (type)
M
M
M'
M
P.M.
H cremicolor Bres.
P,M
P,M
269
1772
2972
160
2151
H cystidiatum Boidin & GiUes
M paratype
P
P
3086
3087
H geogenium (Bres.) J.Erikss.
RM 2052
NH3688
NH9637
PDD 12704
PDD 3855
PDD 12702
PDD 12703
PDD 78974
NH15133,
PDD 78980
NH3406
NH11149
LY5892
LY 5893-PS
MUCL 32103
LY 6228-PS
MUCL 32104
NH10910
Locality
Sweden, Torne Lappmark
Sweden, Dalsland
New Zealand, Bay of Plenty
New Zealand, Bay of Plenty
New Zealand, Bay of Plenty
New Zealand, Bay of Plenty
New Zealand, West Coast
New Zealand, West Coast
Denmark, Lolland
Spain, Gomera
Central African Republic,
Boukokok
Central African Republic,
Bede
Gabon, Libreville
Sweden. Vastersotland
Substratum
Betula
Pinus
Pinus
Pinus
Pinus
Pinus
Undet. wood
Hardwood
Abies
Decid. wood
Bambusa
Bambusa
Elaeis
Conif. wood
GenBank no.
AF429422
AF429423
DQ309071
AF429425
AF429424
DQ658163
DQ658164
AF429426
H. lundeUii (Bourdot) J.Erikss.
M n/a
H. lyndoniae (D.A.Reid) Hjortstam
Sweden Picea AY781277
RM
RM'
2979
3029
H polonense (Bres.) Strid
P,M 1538
P, U 2262
P.M. 2675
U 809
M 942
M 1536
M 2469
H punctulatum (Cooke) J.Erikss.
P, M 938
P;M 1362
P,;M 2833
P,M 1921
H subrigenscens Boidin
P,M 1966
H wakefieldiae (Bres.) J.Erikss.
P.M. 1709
R M 2383
NH15051,
PDD 78906
NH15126,
PDD 78973
NH9061
NH11337
NH12965
NH7507
TH23
NH9331
NH12117
NH7815
NH5175
NH 14069
NH10290
NH 10421
NH9509
NH12107
New Zealand, West Coast
New Zealand West Coast
Romania, Iasi
Turkey, Trabzon
Russia, Krasnodar
Canada, Ontario
Norway, Nord Trondelag
Romania, Brasov
Russia, Krasnodar
Scotland, Perthshire
Sweden, Dalsland
USA, North Carolina
Denmark, Jylland
Denmark, Jylland
Finland, Etela-Hame
Russia. Krasnodar
Weinmannia
Undet. wood
Decid. wood
Fagus
Fraxinus
Decid. wood
Ulmus
Decid. wood
Abies
Betula
Fagus
Decid. Wood
Betula
Pinus
Abies
DQ309070
DQ309069
DQ309066
DQ309065
DQ309067
AF429408
AF429411
AF429409
AF429410
AF429427
AF429419
AF429416
(con
142 New Zealand Journal of Botany, 2007, Vol. 45
Table 1 (continued)
Taxon FCUG no. Collection no.
H. zealandicum (G.H.Cunn.) Hjortstam
P, M 3009
M
M
M
M
M
M
OUTGROUP
Hyphoderma setigerum (Fr.) Donk
P 476
NH15340,
PDD 79138
NH15106,
PDD 78955
NH15339,
PDD 79137
PDD 12433
PDD 12711
PDD 70457
BCP643,
PDD 85560
NH6748
Hyphoderma definitum (H.S.Jacks.) Donk
P 2426
Antrodiella romellii
P
Steccherinum ochraceum
P
NH12266
Renvall
3501(KUO)
Ryvarden
18315(0)
Locality
New Zealand, Hamilton
New Zealand, West Coast
New Zealand, Hamilton
New Zealand, Rangitikei
New Zealand, Waikato
New Zealand, Mid Canterbury
New Zealand, Bay of Plenty
Canada, BC
Russia, Krasnodar
Finland, Lammi
Norway, Vestfold
Substratum
Undet. wood
Undet. wood
Undet. wood
Dacrydium
Podocarpus
Nothofagus
Undet. wood
Abies
Abies
Prunus
Fagus
GenBank no.
DQ309068
AJ534259
AJ534293
AF126902
AF126906
MAFFT version 5.667 (Katoh et al. 2005) and then
manually adjusted in the editor of Seaview (Galtier
etal. 1996). Sequences aligned well within each of
six groups corresponding to the major clades shown
in Fig. 1, but the alignments across all groups neces-
sitated the introduction of gaps. Ambiguous align-
ments were excluded (nucleotide positions 109-146,
472-478) from the analyses.
A Bayesian phylogenetic analysis was carried
out in MrBayes 3.0 (Ronquist & Huelsenbeck 2003)
with best-fit models of nucleotide evolution for
ITS1, 5,8S, andITS2 estimated by MrModeltest2.2
(Nylander 2004). Eight default-setting Metropolis-
Coupled Markov Cham Monte Carlo (MCMCMC)
chains were run for 10 million generations with
trees sampled every 5000 generations and an initial
burn-in of 50%. After discarding the trees prior to
the burn-in threshold, a 50% majority-rule consensus
phylogram was computed from the remaining 1000
trees.
A heuristic parsimony search was set up in PAUP
4.0bl0 (Swofford 2003) with 100 000 random
addition sequence replicates, each holding 5 trees per
step. TBR branch swapping was employed with the
MULTREES option turned on. Clade support was
estimated in PAUP through 25 000 replicates of 37%
JAC jackknife (Farris et al. 1996), each employing
5 rounds of random addition sequence with 2 trees
held per step and TBR swapping with at most 2
trees saved per replicate. Genetic distance (uncor-
rected "p") was also calculated in PAUP (Swofford
2003). GenBank numbers and specimens examined
are provided in Table 1. The alignment matrix can
be accessed through TreeBASE (study accession
number = S 1504).
The macro and micromorphology of specimens
were studied under a dissecting microscope (xl2)
and light microscope (xl250) using phase contrast
and bright field optics. Squash m ounts were prepared
in 2% KOH. Spore measurements were undertaken
from spore prints where available and from basidi-
omata in the remaining cases. For each specimen,
the length and width of 30 spores, 10 basidia, and
10 cystidia were recorded.
Paulus et al.—Hypochnicium from New Zealand 143
Fig. 1 The consensus phylogram
from the Bayesian phylogenetic
inference. Supportvalues are given
asBayesianPosterior Probabilities/
jackknife values from the parsi-
mony analysis (support values
above 0.5/50 are given).
— Artrodiella romelliU AF126%2)
Stecchermum ochraceum (AF12&906)
Hyphodema setigerum (AJ534250)
1 0 / 1 0 0
Hyphoderma definitum (AJ534293)
— Hypochnicium lundellu (AY7S1277)
I Hypochniciumlyndomae (DQ309069)
1 0/1001 Hypochnicium lyndomae(DQ309070)
Nodotia
Hypochnicium zealandicum (DQ309068)
Hypochnicium polonense (DQ309065)
Hypochnicium polonense (DQ309066)
• Hypochnicium polonense (DQ309067)
- Hypochnicium subrigescens (AF429427)
Hypochnicium geogenium (AF42942S)
I Hypochnicium wakefieldiae (AF429416)
Hypochnicium wakefieldiae (AF429419)
Hypochnicium albostramineum (AF429423)
1 0 /100 L Hypochnicium albostramineum (AF429422)
Hypochnicium punctulatum (AF429411)
Hypochnicium punctulatum (AF429409)
Hypochnicium punctulatum (AF429408)
Hypochnicium punctulatum (AF429410)
- Hypochnicium cystidiatum (DQ&581&4)
Hypochnicium cystidiatum (DQS5SISJ)
Hypochnicium aotearoae (DQ309071)
Hypochnicium cremicolor(AF429425)
Hypochnicium cremicolor(AF429424)
1 0 /100
1 0 / 9 2
0 8 1 / 6 8
RESULTS
Molecular analysis
Homology establishment was difficult for 45 base
pairs, which were excluded from further analyses.
The final alignment consisted of 26 taxa and 625
aligned positions, 347 of which were constant, 53 of
which were variable but parsimony uninformative,
and 225 (36%) of which were parsimony informa-
tive. As suggested by MrModeltest, the nucleotide
evolution models GTR+G (IT. SI), K80 (5.8S), and
GTR+I+G (ITS2) were employed in the heteroge-
neous Bayesian analysis. Convergence and chain
mixing was examined a posteriori and found to be
satisfactory. A 50% majority-rule consensus phylo-
gram of the Bayesian analysis is shown in Fig. 1. The
parsimony analysis returned 15 most parsimonious
trees of 706 steps (CI = 0.6190, RI = 0.7988). The
supportvalues from thejackknife analysis are super-
imposed onto the Bayesian tree in Fig. 1.
The results of the Bayesian and the parsimony
analyses were highly concordant. Hypochnicium is
identified as monophyletic with respect to the out-
groups (1.0 Bayesian Posterior Probabilities (BPP),
81% jackknife), and most of its terminal clades
are strongly to very strongly supported. Morpho-
logically similar members of the H. punctulatum
complex, characterised by ornamented spores, form
144 Hew Zealand Journal of Botany, 2007 ?Vol 45
S5D12704 Scalebar = 20mm
the H pi&Ktuktii&n ckde in the phylogenetic m&fy-
ses Inthe parsmony ana^sE ? i f p&£2ctukE&&n is
we zbfy supported (71%1 as a sister clade of the
H wt2$%fl£$t&2£fl£ QEKNtrtE?i2n£&&?i clade ? but flus
clade is not re co ̂ used in the Bayesian ana^ sis ?
which places these two clade s to geflier with the H
^oup ni trichotomy HypochincJLM
(previous ̂ reporte d from Hew Zealand
by Curaur^LamllSfiSl as C^fi^^m putf fij&^ml E
most closed associate d w i h H cremico$&r, and H
^d iddu f l The remaining 1hre e clades lie lide taxa
with smooth spores £&p\^fr^2Lm poS&nenxe and
if z£okm&c&&n form a clade |"G>^L^toKpj2^^? as
do if tyn&?mQ£ and if km&Hs Despite posse ssing
smooih spores ? i f su&ngExenx and if
i with the if p&£Kb&E&&n clade (Fig 1]
TAXONOMY
E an easi^ rec o^used genus char-
acterise dby fluct;-wale d? cyanophious spores and
cbstmcthyphae Julich (19791 erected the genus
Q>i?ep^^zcpj2J to accommodate ff &okm&ca7 a
single species of 'tesupinate basidiomycetes with
thrk-walled spores ?yelliWish-brown lamprocystid-
la tthict:-walled ? encruste d cystiiial andrather large
yellowish-brown?thick- walled basal hyphae" An-
other species, G poSomnxe? was later added by
Stagers & Buchanan (19911 G ^ ^ ^ I ^ J J J was
synonymise d with i&pL^^^JLra by Hjortstam
(1997] without further comment, this synonymy
is confiimed here The raflier thick-walled? cyano-
philous spores as well as its distinct hyphae unite
G>"?eptoKpj3J with specie s of i j p ^ f r n ^ o a Both
genera also form a jointmonophyletic ^mip nthe
ITSsecpience anat^sis (T^ 1]
The genus J^£\i^L2was mtroduce dby Hjortstam
(19S71 to ace cmnodate AT oxpera, apseudodimitic
Jfypm ^ i n Lra - lik e sf̂ c le s wifti thr k- wait d a cule al
etments Later AT L25¥?uwasrec o^use d as a taxo-
nomic synonym of Q&n£it2 tyn&mxie D A Re id?
and Hjortstam (19971 fonne d anew c ombmatin in
I$>pocfoEt2&&?i The genus M\£fta was later re intro-
duced to accommodate JV tyn&?mQ£ and a simihr
species?AT jwaffl^ (Lopea A M ^ i t l Hjortstam A
Eyvarden (Hjortstam * Eyvarden 2 004] Hiylo-
genetic ana^ses of the ITS re g»n phce s AE\i^a
^rai^a^wihinthe genus ijp\^^E^iJfl (F^ l]as
asistertaxonofif JiniJSj (Eourd 1 JEtikss As the
type secies of j^£\£tois nchidedwithinijp\^^E-
?the genus AE\i^a is phce din synon>my wih
The earlier synonyms
AReid] Hjortstam and if
SE Lopea A J E TOj^it are herewith re instated for
AT tyn&?mQ£ and AT JCTS ̂ :2 ?respe ctive^
Pauiis ? Hilsson A
Halinb ?sp nov Fig 2,3
DidotH-ost: Basidioma re supinatum, tenuiter mem-
branulaceum, hypochnoideum, albidom ve 1 ebur-
neum Systema hyphale monomiricum Cystidia
pnrnana me bsa? subcylindri: a? 100-2CO * 7-10 ^im?
plus minusve homogenea Cystidia secundaria
Paulus et al.—Hypochnicium from New Zealand 145
eminentia vel inclusa, subcylindrica vel obpyriformia,
60-120(-140) x 11-16 um, parietibus incrassatis,
1.6-2.4 umcrassis. Basidia24-32 x 5.6-8.8 um, tet-
rasporia. Sporae ovoideae vel subglobosae, 5.6-8.7
x 4.8-7.3 um, verrucosae, tunica crassa.
HOLOTYPUS: New Zealand, Bay of Plenty, Pinedale
near Putaruru, on fallen wood of Pinus radiata,
J. M. Dingley s.n., Nov 1953, PDD 12704.
ETYMOLOGY: Aotearoa ("land of the long white
cloud") is a commonly used Maori designation for
New Zealand. In Latin it must be treated as a femi-
nine noun of the first declension, hence the genitive
epithet aotearoae.
DESCRIPTION: Basidiome membranaceous, resupi-
nate, effused, hypochnoid, white to cream when
dry, 0.1-0.3 mm thick, hymenium drying smooth,
margin not especially differentiated. Hyphal system
monomitic, hyphae hyaline, clamped; basal hyphae
distinctly thick-walled, 4-9 um wide. Subiculum
varying with age and growth conditions, loosely
woven when young to dense when mature. Cystidia
of two kinds are present, which vary in relative
abundance in different fruiting bodies: subcylindri-
cal cystidia, 100-200 x 7-10 um, enclosed in mature
fruiting bodies, thin to slightly thick-walled, staining
strongly in 1% phloxine; distinctly thick-walled
cystidia, 60-120(-140) x 11-16 um, projecting or
enclosed, subcylindrical or with inflated bases, walls
1.6-2A urn thick, cystidial apex not encrusted in
young specimens, in some older specimens a dark
cap of fine crystalline material present. Basidia nar-
rowly clavate to sinuous, 24-32 x 5.6-8.8 um, 4-
sterigmate and with a basal clamp. Spores ovoid
to subglobose, 5.6-8.7 x 4.8-7.3 um (mean 7.9 x
6.7 um), thick-walled, verrucose, cyanophilous.
HABITAT: On decaying wood.
SPECIMENS EXAMINED: NEW ZEALAND: BAY OF
PLENTY: Rotorua State Forest, on fallen wood
of Pinus ponderosa, R. Murray, Oct 1932, PDD
3855; Atiamuri, on Pinus radiata wood and bark,
J. M. Dingley, Nov 1953, PDD 12702; Atiamuri,
on Pinus radiata wood and bark, J. M. Dingley,
Nov 1953, PDD 12703. WEST COAST: Jackson
Bay, on fallen log of undetermined hardwood,
N. Hallenberg, 5 Apr 2004, PDD 78980; Jackson
Bay, on fallen wood, N. Hallenberg, 5 Apr 2004,
PDD 78974.
NOTES: New Zealand specimens are morphologi-
cally similar to H cystidiatum as they present two
types of cystidia: elongated, subcylindrical cystidia
and shorter, thick-walled ones. The morphological
similarity, however, is not reflected in the phylogram
Fig. 3 Hypochnicium aotearoae PDD 12704. A, Cystidia
of two kinds: thin-walled elongate and thick-walled subcy-
lindrical or with inflated bases. B, Basidia and thick-walled,
verrucose spores. Scale bars: A = 20 |xm; B = 10 |xm.
based on ITS sequence data (Fig. 1; see Discussion).
The cystidia of H. aotearoae are on average larger
and more robust with thicker cystidial walls. In addi-
tion, some cystidia in African specimens bear crystal
encrustations, which were not observed in freshly
collected New Zealand material. Some older New
Zealand specimens showed black encrustations at
the cystidial apex, which were difficult to interpret
and may be artefacts of the drying process.Spore
sizes in H. aotearoae vary considerably and fall in
the range of those of H. punctulatum whereas H.
cystidiatum has smaller spores resembling those of
H. cremicolor (Fig. 4). The presence of thick-walled
basal hyphae indicates a relationship of both H.
cystidiatum and H. aotearoae with H. wakefieldiae
(Eriksson & Ryvarden 1976; Nilsson & Hallenberg
2003) but these have also, to some extent, been ob-
served in specimens of H. cremicolor in this study.
In contrast, they were not detected in specimens of
H. punctulatum oxH. albostramineum.
146 New Zealand Journal of Botany, 2007, Vol. 45
8.0
E 7.0
5.0
H. punctulatun}.,--
H.aotearoae.,,"'
. * • •
. ^
\ H. albostramineum
H. mkefieldiae
o H. albostramineum
n H. punctulatum
a H. wakefieldiae
o H. cremicolor
A H. aotearoae
• H. cystidiatum
H. cremicolor
5.5 6.5 7.5 8.5
Spore length (urn)
9.5 10.5
Fig. 4 Mean spore dimensions («
= 30) for representatives of the H.
punctulatum complex.
Hypochnicium lyndoniae (D.A.Reid) Hjortstam,
Mycotaxon 54, 187 (1995)
BASIONYM: Odontia lyndoniae D.A.Reid, Kew Bul-
letin of Miscellaneous Information 10, 641 (1956).
= Nodotia aspera Hjortstam, Mycotaxon 28, 33
(1987).
=Nodotia lyndoniae (D.A.Reid) Hjortstam, Synopsis
Fungorum 18, 18 (2004).
NOTES: This species was described from Australia
and recorded by Cunningham (1959) for New Zea-
land. Although it forms skeletocystidia, it belongs
in Hypochnicium on the basis of its thick-walled,
cyanophilous spores. Its position within Hypoch-
nicium is also confirmed by phylogenetic analysis
of the ITS region (Fig. 1). H. gomezii S.E.Lopez &
J.E.Wright differs from H lyndoniae primarily in
spore size (Hjortstam & Larsson 1994).
Hypochnicium zealandicum (G.Cunn.) Hjortstam,
Windhalia 23, 3 (1998). Fig. 5
= Pellicularia zealandica G.Cunn., Transactions of
the Royal Society of New Zealand Si, 322 (1953).
= Botryobasidium zealandicum (G.Cunn.) Boidin,
Cahiers de laMaboke 8, 25 (1970).
= Gyrophanopsiszealandica (G.Cunn.) Julich, Per-
soonia 10, 329 (1979).
NOTES: The specimens of H zealandicum examined
are morphologically similar to H polonense. Stalp-
ers & Buchanan (1991) studied the type specimen of
Pellicularia zealandica and found the encrustation
of the cystidia to be darker than in H polonense.
This observation could not be confirmed for the
recently collected specimens we examined. The
only other distinguishing characters reported were
slightly smaller and more broadly ellipsoid spores in
G. zealandica (Stalpers & Buchanan 1991; Boidin
& Gilles 2000). However, we detected an overlap in
spore ranges for specimens from New Zealand and
the Northern Hemisphere (Fig. 6). As specimens of
H zealandicum and/f. polonense cannot be reliably
separated by morphology alone, both taxa could be
placed in synonymy. However, the degree of genetic
differentiation in phylogenetic analyses suggests
that/f. zealandicum may represent a separate taxon.
Currently, we choose to retain H zealandicum as a
separate species in the light of the phylogenetic and
geographic information available.
Gloeohypochnicium analogum (Bourdot &
Galzin) Hjortstam, Mycotaxon 28, 30 (1987)
BASIONYM: Gloeocystidium analogum Bourdot &
Galzin, Societe de Mycologique de France Bulletin
28, 366 (1913).
= Corticium globososporum G.Cunn., Transac-
tions of the Royal Society of New Zealand 82, 285
(1954).
= Hypochnicium analogum (Bourdot & Galzin)
J.Erikss., Symbolae botanicae Upsalienses 16, 101
(1958).
NOTES : This species has been placed in the genus Hy-
pochnicium because of its verrucose, cyanophilous
spores. However, in scanning electromicrographs the
ornamentation was shown to differ from other spe-
cies within the genus (Eriksson & Ryvarden 1976).
The presence of true gloeocystidia or sulfocystidia
and its phylogenetic position outside Hypochnicium
based on LSU sequence data (Larsson & Larsson
2003; Binder et al. 2005) support the erection of a
monotypic genus for this taxon (Hjortstam 1987).
147
fpecie* re corded from New Zealand
GSoeofypoc fastis
Key to the fpecie* of Hypochm&iim and
j1 I^menophore odontoidwithpenicilhte
I^menophore smooih
Crloe ocystidn present, daitening in sulfo-vaiull^
Crloe ocystidn a.b sent, other cystidia. present
3 One type of cystiiiim pre sent, septate with c lamps ? thin to thrk-walle d frralls up to 1
cyhndrE al or tapenng, spore s smooth
Two types of cystidia. present, both aseptate ? one thin-wile d and subcylindnca.1, the oiher
thick-walled frra.lt up to 2 4 \sm 1hickl? subcylindnc al or with inflated base ? spores venucose
•
thick],
DISCUSSION
The n ices t level of interspe cifc genetic divergence
recorded n J$>pocfoRC2&&n species was 0 25?for
exanple ?betwe en 1he two Hew Ze aland spec K s if
L^jfiLiSiJfl and if km&&2 Thr level of divergence
E hi^ierthan nterspe cifc genetn drtance srepoited
for some other ba.siiicanyc etes ? which ranged, for
example ̂ between 0 05 and 0 12 nthe genus Scfezo-
fCTu(Pnilis etal 2000] Despite the hi^ilevel of
divergence between some taia ji^logenetic ana.^-
sis of ITS secpience data, supports the hypothesis
that i&p\^^Efaum is a. monophyletic ^m^i with
respect to the out^mips |Flg 1] Thr coiTobora.tes
the view of Eriksson & ^varden (197o"lba.sed on
moiphological studies that in spite of 1he presence
of some drtinct sub^m^s ? JfypocfoRcu&n isLLea.s-
1^ rec o^uae d and ra.ftier well separatedfrom other
genera." Some moipho logic a. 1 characters appear to
be good syna.pomoiphies dhrk-walled spores ? dis-
tiict,nch^branchedsubicularhypha.el?whii oftier
prominent chara.cters are not (pre sence and type of
cystidia.? structure of hymenialsurfic e] Asana.ter-
native hypoftie SE itis possible to bok upon Gyroph-
L^Kpjjj andAE\i^a as cbsnnctgenera. However^io
uninngmoiphobgica.lchara.cters c ouHbe lientuied
for the apparent^ c bse ^ r e htedUxa. H fymtemtx
and H £ j i i j ^ Iti 4dditnn?this stuc^ suggests that
the type sp ecie s of JfypocfoRcu&n ? H
(Somneif A ft] JbhnErikss
be ne sted in the cla.de c onta.inrig AE\i^a and Gy-
?pp^^Kpj2j? a.s i f iu i iJSjhas been prop ose d to
be c lose^ re late d to if ^CTa^riLm (Eriksson A
^varden 1976] One orb o1h of ftie se genera.would
then be a. younger synonym of JfypocfoRcu&n, and
148 New Zealand Journal of Botany, 2007, Vol. 45
7.0
6.0
E
1 5.0
a
CO
4.0
3.0
H. lyndoniae
H. zealandicum
H. polonense
4.0 5.0 6.0 7.0
Spore length (pm)
8.0
Fig. 6 Mean spore dimensions
(n = 30) for representatives of H.
polonense, H. zealandicum, and
H. lyndoniae.
c H. polonense
• H. zealandicum
A H. lyndoniae
9.0
the remaining species in Hypochnicium would be
left without a well-recognised name.
In contrast to fruitbody and cystidial morphology,
spore ornamentation appears to be an important
synapomorphy for part of the species included in
this study (Fig. 1). Differences in wall thickness of
hyphae and septation of cystidia, as observed, for
example, in H. polonense and H. lyndoniae, appear
of importance only for species but not generic de-
limitation. In contrast, the presence of gloeocystidia
in Gloeohypochnicium is an important diagnostic
character for generic delimitation as phylogenetic
analyses of the large subunit of ribosomal DNA
placed this taxon on a different clade from Hypoch-
nicium (Binder et al. 2005).
Morphological and molecular analyses agree
in delimiting species of Hypochnicium as sepa-
rate taxa, but they do not provide congruent results
with respect to the evolutionary relationships for all
taxa included in the present study. For example, H.
cystidiatum and H. aotearoae share rather distinct
morphological characters, which may indicate a
close relationship. These include, for example, the
presence of two types of cystidia and a high degree
of similarity in their thick-walled cystidia. However,
phylogenetic analysis places them in separate clades
(Fig. 1). Conversely, H. lyndoniae and H. lundellii
differ considerably in their morphology despite their
apparently close phylogenetic relationship. Hypoch-
nicium lyndoniae has been described as "pseudodi-
miticwith thick-walled skeletocystidia" (Hjortstam
1987; Hjortstam & Ryvarden 2004) while H. lundel-
lii is clearly monomitic and does not possess any
cystidia. It cannot be excluded that the results of ITS
sequence analysis may be distorted, for example, by
nonrandom substitution events in ITS sequences,
well documented in other organisms and fungal
species (e.g., Levinson & Gutman 1987; Platas et
al. 2001). Analyses of additional genomic regions
may clarify this incongruence in the future.
Morphological analyses indicate that H. zea-
landicum could be accommodated within the cur-
rent broad morphological species concept of H.
polonense. Hypochnicium polonense is a species
with a worldwide distribution that has a remarkable
variation of spore shapes and dimensions, often in
a single collection (Eriksson & Ryvarden 1976).
The rather heterogenous nature of H. polonense is
also recognised in the presence of two intersteril-
ity groups from the Northern Hemisphere, which
cannot be separated on the basis of morphology
(Hallenberg 1991). However, H. zealandicum is cur-
rently retained as a separate species because of the
apparent genetic differentiation of the single culture
that was available from New Zealand compared with
H. polonense from other parts of the world (Fig. 1).
Further molecular studies are required to establish
which taxonomic entities within this morphological
group are present in the Pacific region. Although
New Zealand's geographic isolation may be a factor
contributing to speciation in some instances, stud-
ies of other species have indicated that gene flow
may occur across great distances. For example,
ITS sequence analysis of New Zealand, Australian,
and North American cultures of Schizopora radula
showed little genetic differentiation across that range
(Paulus et al. 2000).
In conclusion, the current study supports a wide
delimitation of the genus Hypochnicium, which is
characterised by thick-walled, cyanophilous spores
and distinct, richly branched subicular hyphae. The
striking differentiation in cystidia of some species
Paulus et al.—Hypochnicium from New Zealand
is important only at the level of species delimitation
with the exception of gloeocystidia. Three species of
Hypochnicium have been reported from New Zea-
land, and of these, H. zealandicum and H. aotearoae
are endemic as far as known. Their genetic dif-
ferentiation from other morphologically similar
taxa indicates that there is a need to reassess the
status of New Zealand corticioid taxa (Cunningham
1963) currently referred to by Northern Hemisphere
names.
ACKNOWLEDGMENTS
We are much indebted to Vivian Alden for technical
assistance. This study was financially supported through
research grants to NH from the Vidfelt Foundation
and the Royal Academy of Arts and Sciences in
Goteborg, both of which are gratefully acknowledged.
BP gratefully acknowledges the support of a Landcare
Research capability grant. We thank E. Ljungstrand and
S. Pennycook for comments on the Latin diagnosis. The
curators of LY and MUCL have kindly placed material
at our disposal.
REFERENCES
Binder M, Hibbett DS, Larsson K-H, Larsson E, Langer
E, Langer G 2005. The phylogenetic distribution
of resupinate forms across the major clades of
mushroom-forming fungi. Systematics and Bio-
diversity 3: 113-157.
Boidin J, Gilles G 1971. Basidiomycetes Corticiaceae de la
Republique Centrafricaine III. Le genre Hypoch-
nicium. Cahiers de la Maboke 9: 89-93.
Boidin J, Gilles G 2000. Basidiomycetes Aphyllopho-
rales de l'île de la Reunion. XX — Le genre
Hypochnicium Eriksson. Bulletin de la Societe de
Mycologique de France 116: 159-172.
Cunningham GH 1953. Thelephoraceae of New Zealand.
Part II: the genus P ellicularia. Transactions of the
Royal Society of New Zealand 81: 321-328.
Cunningham GH 1959. Hydnaceae of New Zealand. Part
II. The genus Odontia. Transactions of the Royal
Society of New Zealand 86: 65-103.
Cunningham GH 1963. The Thelephoraceae of Australia
and New Zealand. DSIR Bulletin 145: 1-359.
Eriksson J 1958. Studies in the Heterobasidiomycetes and
Homobasidiomycetes—Aphyllophorales of Mud-
dus National Park in North Sweden. Symbolae
botanicae Upsalienses 16: 1-172.
Eriksson J, Ryvarden L 1976. The Corticiaceae. Volume
4. Norway, Oslo, Fungiflora.
Farris JS, Albert VA, Kallersjo M, Lipscomb D, Kluge
AG 1996. Parsimony jackknifmg outperforms
neighbor-joining. Cladistics 12: 99-124.
Galtier N, Gouy M, Gautier C 1996. SEAVIEW and
PHYLO_WIN: two graphic tools for sequence
alignment and molecular phylogeny. Computer
Applications in the Biosciences 12: 543-548.
Gardes M, Brims TD 1993. ITS primers with enhanced
specificity for basidiomycetes —application to the
identification of mycorrhizae and rusts. Molecular
Ecology 2: 113-118.
Hallenberg N 1991. Speciation and distribution in Corti-
ciaceae (Basidiomycetes). Plant Systematics and
Evolution 177: 93-110.
Hjortstam K 1987. Studies in tropical Corticiaceae (Ba-
sidiomycetes) VII. Specimens collected by Leif
Ryvarden from East Africa II. Mycotaxon 28:
19-37.
Hjortstam K 1997. A checklist to genera and species of
corticioid fungi (Basidiomycotina, Aphyllopho-
rales).Windahlia 23: 1-54.
Hjortstam K, Larsson K-H 1994. Annotated check-list
to genera and species of corticioid fungi (Aphyl-
lophorales, Basidiomycotina) with special regards
to tropical and subtropical areas. Windahlia 21:
1-75.
Hjortstam K, Ryvarden L 2004. A re-evaluation of No-
dotia Hjortstam (Corticioid fungi, Basidiomy-
cotina, Aphyllophorales). Synopsis Fungorum
18: 17-19.
Holmgren PK, Holmgren NH 1998 onwards (continuously
updated). Index Herbariorum. New York Botanical
Garden. http://sciweb.nybg.org/science2/Index-
Herbariorum.asp [accessed 23 Nov 2005].
Julich W 1979. Studies in resupinate Basidiomycetes
— VI. Persoonia 10: 325-336.
Katoh K, Kuma K, Toh H, MiyataT 2005. MAFFT version
5: improvement in accuracy of multiple sequence
alignment (describes the G-INS-i, L-INS-i and
E-INS-i strategies). Nucleic Acids Research 33:
511-518.
Larsson E, Larsson K-H 2003. Phylogenetic relation-
ships of russuloid basidiomycetes with em-
phasis on aphyllophorales taxa. Mycologia 95:
1037-1065.
Larsson K-H, Larsson E, Koljalg U 2004. High phyloge-
netic diversity among corticioid homobasidiomyc-
etes. Mycological Research 108: 983-1002.
Levinson G, Gutman GA 1987. Slipped-strandmispairing:
a major mechanism for DNA sequence evolution.
Molecular Biology and Evolution 4: 203-221.
Nilsson RH, Hallenberg N 2003. Phylogeny of the Hy-
pochnicium punctulatum complex as inferred from
ITS sequence data. Mycologia 95: 54-60.
149
150 New Zealand Journal of Botany, 2007, Vol. 45
Nylander JAA 2004. MrModeltest v2. Program distrib-
uted by the author. Evolutionary Biology Centre,
Uppsala University.
Parmasto E, Nilsson H, Larsson K-H 2004. Cortbase
version 2. Extensive updates of a nomenclatural
database for corticioid fungi (Hymenomycetes).
Phyloinformatics 5: 1-7.
Paulus B, Hallenberg N, Buchanan PK, Chambers GK
2000. Aphylogenetic study of the genus Schizopo-
ra (Basidiomycota) based on IT S DNA sequences.
Mycological Research 104: 1155-1163.
Platas G, Acero J, Borkowski JA, Gonzalez V, Portal MA,
Rubio V, Sanchez-Ballesteros J, Salazar O, Pelaez
F 2001. Presence of a simple tandem repeat in
the ITS 1 region of the Xylariales. Current Micro-
biology 43: 43-50.
Reid DA 1956. New or interesting records of Australasian
Basidiomycetes. Kew Bulletin 10: 631-647.
Ronquist F, Huelsenbeck JP 2003. MRBAYES 3: Baye-
sian phylogenetic inference under mixed models.
Bioinformatics 19: 1572-1574.
Stalpers JA 1985. Type studies of the species of Corticium
described by G.H. Cunningham. New Zealand
Journal of Botany 23: 301-310.
Stalpers JA, Buchanan PK 1991. Type studies of the spe-
cies of Pellicularia and Peniophora described
by G.H. Cunningham. New Zealand Journal of
Botany 29: 331-340.
Swofford DL 2003. PAUP*. Phylogenetic analysis using
parsimony (*and other methods). Version 4.0bl0.
Massachusetts, Sunderland, Sinauer Associates.