available online at www.studiesinmycology.org
doi:10.3114/sim.2009.64.02
studies in MyCology 64: 17–47. 2009.
phylogenetic lineages in the Capnodiales
P.W. Crous1, 2*, C.L. Schoch3, K.D. Hyde4, A.R. Wood5, C. Gueidan1, G.S. de Hoog1 and J.Z. Groenewald1
1
CBS-KNAW Fungal Biodiversity Centre, P.O. Box 85167, 3508 AD, Utrecht, The Netherlands; 2Wageningen University and Research Centre (WUR), Laboratory of
Phytopathology, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands; 3National Center for Biotechnology Information, National Library of Medicine, National
Institutes of Health, 45 Center Drive, MSC 6510, Bethesda, Maryland 20892-6510, U.S.A.; 4School of Science, Mae Fah Luang University, Tasud, Muang, Chiang Rai 57100,
Thailand; 5ARC – Plant Protection Research Institute, P. Bag X5017, Stellenbosch, 7599, South Africa
*Correspondence: Pedro W. Crous, p.crous@cbs.knaw.nl
Abstract: The Capnodiales incorporates plant and human pathogens, endophytes, saprobes and epiphytes, with a wide range of nutritional modes. Several species are
lichenised, or occur as parasites on fungi, or animals. The aim of the present study was to use DNA sequence data of the nuclear ribosomal small and large subunit RNA genes
to test the monophyly of the Capnodiales, and resolve families within the order. We designed primers to allow the ampliication and sequencing of almost the complete nuclear
ribosomal small and large subunit RNA genes. Other than the Capnodiaceae (sooty moulds), and the Davidiellaceae, which contains saprobes and plant pathogens, the order
presently incorporates families of major plant pathological importance such as the Mycosphaerellaceae, Teratosphaeriaceae and Schizothyriaceae. The Piedraiaceae was not
supported, but resolves in the Teratosphaeriaceae. The Dissoconiaceae is introduced as a new family to accommodate Dissoconium and Ramichloridium. Lichenisation, as well
as the ability to be saprobic or plant pathogenic evolved more than once in several families, though the taxa in the upper clades of the tree lead us to conclude that the strictly
plant pathogenic, nectrotrophic families evolved from saprobic ancestors (Capnodiaceae), which is the more primitive state.
Key words: Ascomycetes, Brunneosphaerella, Capnodiales, DNA sequence comparisons, Mycosphaerella, novel primers, systematics.
Taxonomic novelties: Brunneosphaerella Crous, gen. nov., B. jonkershoekensis (Marinc., M.J. Wingf. & Crous) Crous, comb. nov., B. protearum (Syd. & P. Syd.) Crous,
comb. nov., Devriesia hilliana Crous & U. Braun, sp. nov., D. lagerstroemiae Crous & M.J. Wingf., sp. nov., D. strelitziicola Arzanlou & Crous, sp. nov., Dissoconiaceae
Crous & de Hoog, fam. nov., Hortaea thailandica Crous & K.D. Hyde, sp. nov., Passalora ageratinae Crous & A.R. Wood, sp. nov., P. armatae Crous & A.R. Wood, sp. nov.,
Rachicladosporium cboliae Crous, sp. nov.
INTroDuCTIoN
The Dothideomycetes encompasses plant and human pathogens,
endophytes, saprobes and epiphytes. The class presently
contains two subclasses, namely Pleosporomycetidae and
Dothideomycetidae (Schoch et al. 2006, 2009a). Although
the main orders, Pleosporales and Dothideales correlate with
the presence or absence of pseudoparaphyses and other
centrum characteristics, many orders remain unresolved. The
Dothideomycetidae include the orders Dothideales, Capnodiales
and Myriangiales, which lack paraphyses, pseudoparaphyses and
periphysoids. Based on a multi-gene phylogeny, and the presence
of ostiolar periphyses as possible synapomorphy, the Capnodiales
were recognised as the order incorporating the Capnodiaceae,
Davidiellaceae, Mycosphaerellaceae and Piedraiaceae (Schoch et
al. 2006). However, several studies (Hunter et al. 2006, Crous et al.
2007a, b) showed the Mycosphaerellaceae to be polyphyletic, and
to contain additional variation at the familial level, leading to the
circumscriptions of the Teratosphaeriaceae and Schizothyriaceae.
Crous et al. (2009b, c) again revealed Teratosphaeriaceae to be
too widely deined, including some further unresolved families.
The present study focuses on the Capnodiales, which
is based on the Capnodiaceae, representing a group of leaf
epiphytes associated with honeydew of insects, usually visible as
a black growth on leaf surfaces, fruit and twigs. Members of the
Capnodiaceae form supericial ascomata with fasciculate asci, and
hyaline to dark, septate ascospores. Anamorphs are dematiaceous,
and include mycelial (phragmo- to dictyoconidia), spermatial and
pycnidial synanamorphs (Hughes 1976, Cheewangkoon et al.
2009).
The Mycosphaerellaceae was treated as a family in the
Dothideales by Hawksworth et al. (1995), while Kirk et al. (2001)
introduced a separate order, the Mycosphaerellales for this
family, and Kirk et al. (2008) again placed it in the Capnodiales.
The Mycosphaerellaceae is recognised by having characteristic
pseudothecial ascomata that can be immersed or supericial,
embedded in host tissue or erumpent, having ostiolar periphyses,
but lacking interascal tissue at maturity. Ascospores are hyaline, but
in some cases slightly pigmented (Barr 1987), and predominantly
1-septate, although some taxa with 3-septate ascospores have
been recorded (Crous et al. 2003). Although up to 30 anamorph
genera have been linked to Mycosphaerella (Crous et al. 2000,
2001, 2007a–c, 2009a–c, Crous 2009), recent studies have
shown this to be incorrect, and that the family in fact consists of
numerous genera with morphologically conserved Mycosphaerellalike teleomorphs, and distinct anamorphs (Crous et al. 2007a, b,
2009b, c).
Families tentatively placed in the Capnodiales (Lumbsch
& Huhndorf 2007, Kirk et al. 2008) include epiphytes
(Antennulariellaceae,
Capnodiaceae,
Metacapnodiaceae)
(Hughes 1976), saprobes and plant pathogens (Davidiellaceae,
Dissoconiaceae,
Mycosphaerellaceae,
Schizothyriaceae,
Teratosphaeriaceae) (Aptroot 2006, Crous 2009), and colonisers
or hair shafts of mammals (Piedraiaceae) (de Hoog et al. 2000).
To address the status of the Capnodiales as an order, and the
intrafamilial relationships within this order, DNA sequences of
Copyright 2009 CBS-KNAW Fungal Biodiversity Centre, P.O. Box 85167, 3508 AD Utrecht, The Netherlands.
You are free to share - to copy, distribute and transmit the work, under the following conditions:
Attribution:
You must attribute the work in the manner speciied by the author or licensor (but not in any way that suggests that they endorse you or your use of the work).
Non-commercial:
You may not use this work for commercial purposes.
No derivative works: You may not alter, transform, or build upon this work.
For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. Any of the above conditions can be waived if you get
permission from the copyright holder. Nothing in this license impairs or restricts the author’s moral rights.
17
Crous et al.
the 18S, 5.8S and 28S nrRNA genes were generated for a set of
speciically selected taxa. A further aim was to clarify genera within
these families, and resolve anamorph-teleomorph relationships for
the taxa investigated.
mATerIAlS AND meTHoDS
Isolates
Isolates were selected (Table 1 - see online Supplementary
Information) that are representative of the Mycosphaerellaceae
(Crous 1998, Crous et al. 2004a, c, 2006a, b, 2007a),
Schizothyriaceae (Batzer et al. 2005, 2007), Teratosphaeriaceae
(Crous et al. 2007a, 2008b, c, 2009a–c), Piedraiaceae (Kruys et
al. 2006), Davidiellaceae (Braun et al. 2003, Schubert et al. 2007a,
b), Capnodiaceae (Schoch et al. 2006), as well as numerous other
genera for which the familial relationships have remained unclear,
such as the Phaeophleospora complex (Crous et al. 1997, 2007a,
2009b, c, Andjic et al. 2007), Polythrincium (Simon et al. 2009), the
Dissoconium complex (Crous et al. 2004c, 2007c, 2008b, Arzanlou
et al. 2008b), and several less well-known genera represented
by one or two species only. For fresh material excised leaf spots
bearing ascomata were soaked in water for approximately 2 h, after
which they were placed in the bottom of Petri dish lids, with the top
half of the dish containing 2 % malt extract agar (MEA; Crous et
al. 2009d). Ascospore germination patterns were examined after
24 h, and single-ascospore and conidial cultures established as
described by Crous et al. (1991). Colonies were sub-cultured onto
synthetic nutrient-poor agar (SNA), potato-dextrose agar (PDA),
oatmeal agar (OA), MEA (Crous et al. 2009d), and incubated at 25
°C under continuous near-ultraviolet light to promote sporulation.
Other cultures were obtained from the culture collection of the
Centraalbureau voor Schimmelcultures (CBS-KNAW) in Utrecht,
the Netherlands or the working collection of Pedro Crous (CPC).
DNA isolation, ampliication and molecular phylogeny
Genomic DNA was extracted from mycelium taken from fungal
colonies on MEA using the UltraCleanTM Microbial DNA Isolation
Kit (Mo Bio Laboratories, Inc., Solana Beach, CA, U.S.A.). A
part of the nuclear rDNA operon spanning the 3’ end of the 18S
rRNA gene (SSU), the irst internal transcribed spacer (ITS1), the
5.8S rRNA gene, the second ITS region (ITS2) and the irst 900
bp at the 5’ end of the 28S rRNA gene (LSU) was ampliied and
sequenced as described by Cheewangkoon et al. (2008) standard
for all strains included (Table 1). For selected strains (see Table
1), the almost complete SSU and LSU (missing the irst and last
20–30 nucleotides) were ampliied and sequenced using novel and
previously published primers (Table 2; see below).
Novel primers were designed using a variety of complete
SSU and LSU sequences obtained from the GenBank sequence
database (www.ncbi.nlm.nih.gov/). The selection was not limited
only to fungi belonging to the Dothideomycetes but encompassed
as many as possible full sequences in order to make the primers
as robust as possible. We aimed to keep the melting temperature
(Tm) of the novel primers at 40–45 °C and the GC content to
approximately 50 % to keep them as compatible as possible to
existing published primers. Primer parameters were calculated
using the OligoAnalyzer tool on the web site of Integrated
DNA Technologies (http://eu.idtdna.com/analyzer/Applications/
18
OligoAnalyzer/) with the “Oligo Conc” parameter set at 0.2 mM
and the “Na+ Conc” parameter set at 16 mM. A framework of
existing and novel primers was then aligned onto the sequence
of Magnaporthe grisea (GenBank accession AB026819) to derive
primer positions (Table 2) and evaluate coverage over the gene
regions. These primers were ampliied and sequenced in the
following overlapping sections to cover the almost complete SSU
and LSU for the selected strains (Table 2): SSU1Fd or SSU6Fm
with SSU2Rd, SSU2Fd with SSU3Rd, SSU7Fm with SSU4Rd or
SSU6Rm, SSU4Fd with 5.8S1Rd, V9G or LSU1Fd with LSU3Rd,
LSU8Fd with LSU8Rd, LSU4Fd with LSU5Rd, and LSU5Fd with
LSU7Rd. For some strains (Table 3) it was necessary to add an
additional overlap for SSU4Fd with 5.8S1Rd (using SSU4Fd with
SSU7Rm and SSU8Fm with 5.8S1Rd), for LSU8Fd with LSU8Rd
(using LSU8Fd with LSU3Rd and LSU3Fd with LSU8Rd), and for
LSU5Fd with LSU7Rd (using LSU5Fd with LSU6Rd and LSU6Fd
with LSU7Rd) to complete the gaps due to large insertions.
The internal transcribed spacer regions, as well as all insertions
(Table 3) were excluded from all analyses. Sequence data were
deposited in GenBank (Table 1) and alignments in TreeBASE
(www.treebase.org). Two separate analyses were performed:
The irst using only partial LSU data due to the limited number
of complete LSU sequences available and the second using the
almost complete SSU, 5.8S nrDNA and LSU alignment.
Maximum likelihood analyses (ML) were conducted in
RAxML v. 7.0.4 (Stamatakis 2006) for the partial LSU alignment.
A general time reversible model (GTR) with a discrete gamma
distribution and four rate classes was applied. A tree was obtained
by simultaneously running a fast bootstrap search of 1000
pseudoreplicates (Stamatakis et al. 2008) followed by a search for
the most likely tree. Maximum Likelihood bootstrap value (MLBP)
equal or greater than 70 % are given at the nodes (Fig. 1).
Maximum likelihood analyses (ML) were conducted in RAxML
v. 7.0.4 (Stamatakis 2006) for the almost complete SSU, 5.8S
nrDNA and LSU alignment. A general time reversible model (GTR)
with a discrete gamma distribution and four rate classes was
applied to each partition (SSU, 5.8S nrDNA and LSU). A tree was
obtained by simultaneously running a fast bootstrap search of 500
pseudoreplicates (Stamatakis et al. 2008) followed by a search for
the most likely tree. Maximum Likelihood bootstrap value (MLBP)
equal or greater than 70 % are given at the nodes (Fig. 2).
Taxonomy
Fungal structures were mounted in lactic acid, and 30
measurements (× 1000 magniication) obtained per structure type.
The range obtained is presented, except for spore measurements,
where the 95 % conidence intervals are given with the extremes in
parentheses. Colony colours (surface and reverse) were assessed
after 1–2 wk on MEA at 25 °C in the dark, using the colour charts of
Rayner (1970). All cultures obtained in this study are maintained in
the culture collection of the Centraalbureau voor Schimmelcultures
(CBS-KNAW) in Utrecht, the Netherlands (Table 1). Nomenclatural
novelties and descriptions were deposited in MycoBank (Crous et
al. 2004b). Names for which the taxonomy has not been resolved,
but need to be allocated to another genus, are placed in inverted
commas, e.g. “Mycosphaerella” iridis.
PhylogenetiC lineages in the Capnodiales
Table 2. Details of primers used for this study and their relation to selected published primers. Primer names ending with a "d" denotes a
degenerate primer whereas those ending with a "m" denotes speciic primers designed based on the partial novel sequences generated.
The start and end positions of the primers are derived using Magnaporthe grisea GenBank accession AB026819 as reference in the 5'–3'
direction.
Name
Sequence (5’ – 3’)
orientation
%GC Tm (oC)
Start
end
reference
5.8S1Fd
CTC TTG GTT CBV GCA TCG
Forward
57.4
49.8 – 54.2 – 56.8
2333
2350
This study
5.8S1Rd
WAA TGA CGC TCG RAC AGG CAT G
Reverse
52.3
57.6 – 58.9 – 60.2
2451
2472
This study
F377
AGA TGA AAA GAA CTT TGA AAA
GAG AA
Forward
26.9
40.3
3005
3030
www.lutzonilab.net/primers/
page244.shtml
ITS1
TCC GTA GGT GAA CCT GCG G
Forward
63.2
49.5
2162
2180
White et al. (1990)
ITS1F
CTT GGT CAT TTA GAG GAA GTA A
Forward
36.4
39.0
2124
2145
Gardes & Bruns (1993)
ITS1Fd
CGA TTG AAT GGC TCA GTG AGG C
Forward
54.5
48.0
2043
2064
This study
ITS1Rd
GAT ATG CTT AAG TTC AGC GGG
Reverse
47.6
43.1
2671
2691
This study
ITS4
TCC TCC GCT TAT TGA TAT GC
Reverse
45.0
41.6
2685
2704
White et al. (1990)
ITS4S
CCT CCG CTT ATT GAT ATG CTT
AAG
Reverse
41.7
42.9
2680
2703
Kretzer et al. (1996)
Forward
40.9
40.8
2138
2159
White et al. (1990)
ITS5
GGA AGT AAA AGT CGT AAC AAG G
LR0R
GTA CCC GCT GAA CTT AAG C
Forward
52.6
43.2
2668
2686
Rehner & Samuels (1994)
LR2
TTT TCA AAG TTC TTT TC
Reverse
23.5
28.5
3009
3025
www.lutzonilab.net/primers/
page244.shtml
LR2R
AAG AAC TTT GAA AAG AG
Forward
29.4
30.4
3012
3028
www.lutzonilab.net/primers/
page244.shtml
LR3
GGT CCG TGT TTC AAG AC
Reverse
52.9
40.5
3275
3291
Vilgalys & Hester (1990)
LR3R
GTC TTG AAA CAC GGA CC
Forward
52.9
40.5
3275
3291
www.lutzonilab.net/primers/
page244.shtml
LR5
TCC TGA GGG AAA CTT CG
Reverse
52.9
41.0
3579
3595
Vilgalys & Hester (1990)
LR5R
GAA GTT TCC CTC AGG AT
Forward
47.1
37.8
3580
3596
www.biology.duke.edu/fungi/
mycolab/primers.htm
LR6
CGC CAG TTC TGC TTA CC
Reverse
58.8
43.5
3756
3772
Vilgalys & Hester (1990)
LR7
TAC TAC CAC CAA GAT CT
Reverse
41.2
35.3
4062
4078
Vilgalys & Hester (1990)
LR8
CAC CTT GGA GAC CTG CT
Reverse
58.8
44.3
4473
4489
www.lutzonilab.net/primers/
page244.shtml
LR8R
AGC AGG TCT CCA AGG TG
Forward
58.8
44.3
4473
4489
www.lutzonilab.net/primers/
page244.shtml
LR9
AGA GCA CTG GGC AGA AA
Reverse
52.9
43.6
4799
4815
www.lutzonilab.net/primers/
page244.shtml
LR10
AGT CAA GCT CAA CAG GG
Reverse
52.9
41.6
5015
5031
www.lutzonilab.net/primers/
page244.shtml
LR10R
GAC CCT GTT GAG CTT GA
Forward
52.9
41.6
5013
5029
www.lutzonilab.net/primers/
page244.shtml
LR11
GCC AGT TAT CCC TGT GGT AA
Reverse
50.0
43.9
5412
5431
www.lutzonilab.net/primers/
page244.shtml
LR12
GAC TTA GAG GCG TTC AG
Reverse
52.9
39.4
5715
5731
Vilgalys & Hester (1990)
LR12R
CTG AAC GCC TCT AAG TCA GAA
Forward
47.6
43.7
5715
5735
www.biology.duke.edu/fungi/
mycolab/primers.htm
LR13
CAT CGG AAC AAC AAT GC
Reverse
47.1
38.8
5935
5951
www.lutzonilab.net/primers/
page244.shtml
LR14
AGC CAA ACT CCC CAC CTG
Reverse
61.1
47.6
5206
5223
www.lutzonilab.net/primers/
page244.shtml
LR15
TAA ATT ACA ACT CGG AC
Reverse
35.3
32.5
2780
2796
www.lutzonilab.net/primers/
page244.shtml
LR16
TTC CAC CCA AAC ACT CG
Reverse
52.9
42.1
3311
3327
Moncalvo et al. (1993)
LR17R
TAA CCT ATT CTC AAA CTT
Forward
27.8
31.2
3664
3681
www.lutzonilab.net/primers/
page244.shtml
LR20R
GTG AGA CAG GTT AGT TTT ACC
CT
Forward
43.5
43.6
5570
5592
www.lutzonilab.net/primers/
page244.shtml
LR21
ACT TCA AGC GTT TCC CTT T
Reverse
42.1
41.7
3054
3072
www.lutzonilab.net/primers/
page244.shtml
LR22
CCT CAC GGT ACT TGT TCG CT
Reverse
55.0
46.8
2982
3001
www.lutzonilab.net/primers/
page244.shtml
www.studiesinmycology.org
19
Crous et al.
Table 2. (Continued).
Name
Sequence (5’ – 3’)
orientation
%GC Tm (oC)
Start
end
reference
LSU1Fd
GRA TCA GGT AGG RAT ACC CG
Forward
55.0
41.8 – 44.0 – 46.3
2655
2674
This study
LSU1Rd
CTG TTG CCG CTT CAC TCG C
Reverse
63.2
49.6
2736
2754
This study
LSU2Fd
GAA ACA CGG ACC RAG GAG TC
Forward
57.5
45.5 – 46.5 – 47.6
3280
3299
This study
LSU2Rd
ATC CGA RAA CWT CAG GAT CGG
TCG
Reverse
52.1
48.3 – 49.0 – 49.8
3379
3402
This study
GTT CAT CYA GAC AGC MGG ACG
Forward
57.1
44.7 – 47.4 – 50.2
3843
3863
This study
CAC ACT CCT TAG CGG ATT CCG
AC
Reverse
56.5
49.1
3876
3898
This study
CCG CAG CAG GTC TCC AAG G
Forward
68.4
51.2
4469
4487
This study
CGG ATC TRT TTT GCC GAC TTC
CC
Reverse
54.3
47.4 – 48.7 – 50.0
4523
4545
This study
AGT GGG AGC TTC GGC GC
Forward
70.6
51.6
3357 /
5072
3373 /
5088
This study
GGA CTA AAG GAT CGA TAG GCC
ACA C
Reverse
52.0
48.3
5355
5379
This study
LSU3Fd
LSU3Rd
LSU4Fd
LSU4Rd
LSU5Fd
LSU5Rd
LSU6Fd
CCG AAG CAG AAT TCG GTA AGC G
Forward
54.5
48.1
5499
5520
This study
LSU6Rd
TCT AAA CCC AGC TCA CGT TCC C
Reverse
54.5
48.6
5543
5564
This study
LSU7Fd
GTT ACG ATC TRC TGA GGG TAA
GCC
Forward
52.1
46.0 – 47.4 – 48.8
5943
5966
This study
GCA GAT CGT AAC AAC AAG GCT
ACT CTA C
Reverse
46.4
47.9
5927
5954
This study
CCA GAG GAA ACT CTG GTG GAG
GC
Forward
60.9
51.2
3469
3491
This study
GTC AGA TTC CCC TTG TCC GTA
CC
Reverse
56.5
48.9
4720
4742
This study
GGT AGC CAA ATG CCT CGT CAT C
Forward
54.5
47.9
4882
4903
This study
LSU9Rm
GAT TYT GCS AAG CCC GTT CCC
Reverse
59.5
49.2 – 50.0 – 50.9
4979
4999
This study
LSU10Fm
GGG AAC GTG AGC TGG GTT TAG A
Forward
54.5
48.6
5543
5564
This study
LSU10Rm
CGC TTA CCG AAT TCT GCT TCG G
Reverse
54.5
48.1
5499
5520
This study
LSU11Fm
TTTGGTAAGCAGAACTGGCGATGC
Forward
50.0
49.4
3753
3776
This study
LSU12Fd
GTGTGGCCTATCGATCCTTTAGTCC
Forward
52.0
48.3
5355
5379
This study
NS1
GTA GTC ATA TGC TTG TCT C
Forward
42.1
36.9
413
431
White et al. (1990)
NS1R
GAG ACA AGC ATA TGA CTA C
Reverse
42.1
36.9
413
431
www.lutzonilab.net/primers/
page244.shtml
NS2
GGC TGC TGG CAC CAG ACT TGC
Reverse
66.7
53.8
943
963
White et al. (1990)
NS3
GCAAGTCTGGTGCCAGCAGCC
Forward
66.7
53.8
943
963
White et al. (1990)
NS4
CTT CCG TCA ATT CCT TTA AG
Reverse
40.0
38.2
1525
1544
White et al. (1990)
NS5
AAC TTA AAG GAA TTG ACG GAA G
Forward
36.4
40.1
1523
1544
White et al. (1990)
NS6
GCA TCA CAG ACC TGT TAT TGC
CTC
Reverse
50.0
47.5
1806
1829
White et al. (1990)
Forward
50.0
47.5
1806
1829
White et al. (1990)
LSU7Rd
LSU8Fd
LSU8Rd
LSU9Fm
NS7
GAG GCA ATA ACA GGT CTG TGA
TGC
NS8
TCC GCA GGT TCA CCT ACG GA
Reverse
60.0
50.4
2162
2181
White et al. (1990)
NS17
CAT GTC TAA GTT TAA GCA A
Forward
31.6
34.2
447
465
Gargas & Taylor (1992)
NS18
CTC ATT CCA ATT ACA AGA CC
Reverse
40.0
38.0
887
906
Gargas & Taylor (1992)
NS19
CCG GAG AAG GAG CCT GAG AAA C
Forward
59.1
49.3
771
792
Gargas & Taylor (1992)
NS20
CGT CCC TAT TAA TCA TTA CG
Reverse
40.0
37.3
1243
1262
Gargas & Taylor (1992)
NS21
GAA TAA TAG AAT AGG ACG
Forward
33.3
30.5
1193
1210
Gargas & Taylor (1992)
NS22
AAT TAA GCA GAC AAA TCA CT
Reverse
30.0
36.4
1687
1706
Gargas & Taylor (1992)
NS23
GAC TCA ACA CGG GGA AAC TC
Forward
55.0
45.5
1579
1598
Gargas & Taylor (1992)
NS24
AAA CCT TGT TAC GAC TTT TA
Reverse
30.0
36.2
2143
2162
Gargas & Taylor (1992)
SR11R
GGA GCC TGA GAA ACG GCT AC
Forward
60.0
47.8
779
798
Spatafora et al. (1995)
SR1R
TAC CTG GTT GAT TCT GC
Forward
47.1
38.5
394
410
Vilgalys & Hester (1990)
SR3
GAA AGT TGA TAG GGC T
Reverse
43.8
34.8
696
711
www.biology.duke.edu/fungi/
mycolab/primers.htm
20
PhylogenetiC lineages in the Capnodiales
Table 2. (Continued).
Name
Sequence (5’ – 3’)
orientation
%GC Tm (oC)
Start
end
reference
SSU1Fd
CTG CCA GTA GTC ATA TGC TTG
TCT C
Forward
48.0
46.5
407
431
This study
CTT TGA GAC AAG CAT ATG AC
Reverse
40.0
48.7
416
435
This study
SSU2Fd
GAA CAA YTR GAG GGC AAG
Forward
50.0
47.8 – 50.7 – 53.5
930
947
This study
SSU2Rd
TAT ACG CTW YTG GAG CTG
Reverse
47.2
48.4 – 49.9 – 51.2
974
991
This study
SSU3Fd
ATC AGA TAC CGT YGT AGT C
Forward
44.7
48.4 – 49.5 – 50.5
1389
1407
This study
SSU3Rd
TAY GGT TRA GAC TAC RAC GG
Reverse
47.5
49.0 – 52.5 – 56.0
1397
1416
This study
SSU4Fd
CCG TTC TTA GTT GGT GG
Forward
52.9
50.0
1670
1686
This study
SSU4Rd
CAG ACA AAT CAC TCC ACC
Reverse
50.0
50.3
1682
1699
This study
SSU5Fd
TAC TAC CGA TYG AAT GGC
Forward
47.2
48.9 – 50.1 – 51.2
2037
2054
This study
SSU5Rd
CGG AGA CCT TGT TAC GAC
Reverse
55.6
52.5
2148
2165
This study
SSU6Fm
GCT TGT CTC AAA GAT TAA GCC
ATG CAT GTC
Forward
43.3
49.0
423
452
This study
GCA GGT TAA GGT CTC GTT CGT
TAT CGC
Reverse
51.9
50.1
1707
1733
This study
GAG TGT TCA AAG CAG GCC TNT
GCT CG
Forward
55.8
51.0 – 52.2 – 53.3
1153
1178
This study
CAA TGC TCK ATC CCC AGC ACG
AC
Reverse
58.7
49.5 – 50.8 – 52.1
1921
1943
This study
GCA CGC GCG CTA CAC TGA C
Forward
68.4
52.2
1848
1866
This study
TTA CGT CCC TGC CCT TTG TA
Forward
45.0
42.8
2002
2021
de Hoog & Gerrits van den Ende
(1998)
SSU1Rd
SSU6Rm
SSU7Fm
SSU7Rm
SSU8Fm
V9G
Table 3. Isolates containing group I intron sequences. The insertion positions of these introns are derived using Magnaporthe grisea
GenBank accession AB026819 as reference in the 5'–3' direction.
Isolate
Insertion
between
18S or 28S
nrDNA
Intron size
(bp)
Blast result
Batcheloromyces leucadendri CBS 110892
1559 – 1560
18S nrDNA
350
No signiicant similarity
1820 – 1821
18S nrDNA
399
190/252 of AY545722 Hydropisphaera erubescens 18S nrDNA
4875 – 4876
28S nrDNA
328
211/264 of DQ246237 Teratosphaeria mexicana 28S nrDNA
5424 – 5425
28S nrDNA
538
No signiicant similarity
5538 – 5539
28S nrDNA
383
218/283 of EU181458 Trichophyton soudanense 28S nrDNA
1559 – 1560
18S nrDNA
325
No signiicant similarity
1820 – 1821
18S nrDNA
399
191/254 of AY545722 Hydropisphaera erubescens 18S nrDNA
4875 – 4876
28S nrDNA
328
211/263 of DQ246237 Teratosphaeria mexicana 28S nrDNA
5424 – 5425
28S nrDNA
535
75/90 of DQ442697 Arxula adeninivorans 26S nrDNA
5538 – 5539
28S nrDNA
372
34/36 of GQ120133 Uncultured marine fungus 18S nrDNA
Catenulostroma macowanii CBS 110756
1559 – 1560
18S nrDNA
395
297/379 of DQ848302 Mycosphaerella latebrosa 18S nrDNA
5424 – 5425
28S nrDNA
914
No signiicant similarity
Catenulostroma macowanii CBS 111029
1559 – 1560
18S nrDNA
395
303/379 of DQ848302 Mycosphaerella latebrosa 18S nrDNA
Batcheloromyces proteae CBS 110696
5424 – 5425
28S nrDNA
914
No signiicant similarity
Cercospora apii CBS 118712
1820 – 1821
18S nrDNA
733
288/363 of EU167577 Mycosphaerella milleri 18S nrDNA
Cercospora capsici CPC 12307
1820 – 1821
18S nrDNA
732
287/363 of EU167577 Mycosphaerella milleri 18S nrDNA
Cercospora janseana CBS 145.37
1820 – 1821
18S nrDNA
350
295/365 of EU167577 Mycosphaerella milleri 18S nrDNA
Devriesia staurophora CBS 375.81
3560 – 3561
28S nrDNA
309
No signiicant similarity
Miuraea persicae CPC 10069
1820 – 1821
18S nrDNA
603
399/443 of DQ848342 Mycosphaerella populorum 18S nrDNA
Mycosphaerella latebrosa CBS 652.85
1559 – 1560
18S nrDNA
370
234/296 of DQ848311 Septoria betulae 18S nrDNA
1820 – 1821
18S nrDNA
933
Matches same species
2168 – 2169
18S nrDNA
494
377/449 of DQ848326 Septoria alnifolia 18S nrDNA
4875 – 4876
28S nrDNA
481
No signiicant similarity
missing 5018 –
5019
28S nrDNA
Not present
Not present
www.studiesinmycology.org
21
Crous et al.
Table 3. (Continued).
Isolate
Insertion
between
18S or 28S
nrDNA
Intron size
(bp)
Blast result
5424 – 5425
28S nrDNA
680
No signiicant similarity
5538 – 5539
28S nrDNA
471
No signiicant similarity
1559 – 1560
18S nrDNA
370
231/295 of DQ848310 Septoria betulae 18S nrDNA
1820 – 1821
18S nrDNA
918
Matches same species
2168 – 2169
18S nrDNA
494
377/449 of DQ848326 Septoria alnifolia 18S nrDNA
4875 – 4876
28S nrDNA
480
No signiicant similarity
5018 – 5019
28S nrDNA
417
144/181 of AF430703 Beauveria bassiana 28S nrDNA
5424 – 5425
28S nrDNA
680
No signiicant similarity
5538 – 5539
28S nrDNA
471
No signiicant similarity
Mycosphaerella marksii CBS 110942
1559 – 1560
18S nrDNA
341
332/355 of DQ848296 Mycosphaerella musae 18S nrDNA
Mycosphaerella marksii CPC 11222
1559 – 1560
18S nrDNA
341
332/355 of DQ848296 Mycosphaerella musae 18S nrDNA
Mycosphaerella latebrosa CBS 687.94
Passalora-like genus CPC 11876
5538 – 5539
28S nrDNA
580
No signiicant similarity
Passalora bellynckii CBS 150.49
1559 – 1560
18S nrDNA
409
147/191 of DQ848296 Mycosphaerella musae 18S nrDNA
Passalora dodonaea CPC 1223
5424 – 5425
28S nrDNA
738
No signiicant similarity
Phacellium paspali CBS 113093
4875 – 4876
28S nrDNA
340
161/197 of DQ248314 Symbiotaphrina kochii 28S nrDNA
Phaeophleospora eugeniicola CPC 2557
missing 5424 –
5425
28S nrDNA
Not present
Not present
5538 – 5539
28S nrDNA
744
No signiicant similarity
Phaeophleospora eugeniicola CPC 2558
5424 – 5425
28S nrDNA
1846
No signiicant similarity
5538 – 5539
28S nrDNA
744
No signiicant similarity
Pseudocercospora angolensis CBS 112933
5018 – 5019
28S nrDNA
379
No signiicant similarity
Pseudocercospora angolensis CBS 149.53
5018 – 5019
28S nrDNA
379
No signiicant similarity
Pseudocercospora punctata CBS 113315
5424 – 5425
28S nrDNA
723
No signiicant similarity
5538 – 5539
28S nrDNA
725
67/73 of AF430699 Beauveria bassiana 28S nrDNA
Pseudocercospora punctata CPC 10532
5424 – 5425
28S nrDNA
731
No signiicant similarity
5538 – 5539
28S nrDNA
725
67/73 of AF430699 Beauveria bassiana 28S nrDNA
Ramularia coleosporii CPC 11516
1559 – 1560
18S nrDNA
445
No signiicant similarity
Ramularia grevilleana CPC 656
5538 – 5539
28S nrDNA
546
No signiicant similarity
Septoria apiicola CBS 400.54
5424 – 5425
28S nrDNA
763
No signiicant similarity
Septoria obesa CBS 354.58
1820 – 1821
18S nrDNA
575
No signiicant similarity
2168 – 2169
18S nrDNA
548
394/454 of DQ848326 Septoria alnifolia 18S nrDNA
4875 – 4876
28S nrDNA
430
No signiicant similarity
Septoria pyricola CBS 222.31
5424 – 5425
28S nrDNA
723
No signiicant similarity
Septoria quercicola CBS 663.94
1559 – 1560
18S nrDNA
334
241/308 of DQ848303 Mycosphaerella latebrosa 18S nrDNA
1820 – 1821
18S nrDNA
442
379/452 of DQ848335 Mycosphaerella latebrosa 18S nrDNA
4875 – 4876
28S nrDNA
345
No signiicant similarity
5018 – 5019
28S nrDNA
367
122/155 of DQ518980 Lipomyces spencermartinsiae 28S
nrDNA
5424 – 5425
28S nrDNA
526
No signiicant similarity
5538 – 5539
28S nrDNA
603
No signiicant similarity
Septoria rosae CBS 355.58
1820 – 1821
18S nrDNA
496
No signiicant similarity
Sonderhenia eucalypticola CPC 11252
1559 – 1560
18S nrDNA
408
339/404 of DQ848314 Mycosphaerella populorum 18S nrDNA
4875 – 4876
28S nrDNA
337
229/289 of AB044641 Cordyceps sp. 28S nrDNA
5424 – 5425
28S nrDNA
705
No signiicant similarity
1559 – 1560
18S nrDNA
379
40/44 of AB007686 Exophiala calicioides 18S nrDNA
5018 – 5019
28S nrDNA
376
No signiicant similarity
Stigmina platani CBS 110755
Stigmina synanamorph CPC 11721
5018 – 5019
28S nrDNA
371
No signiicant similarity
Teratosphaeria aff. nubilosa CBS 114419
4871 – 4872
28S nrDNA
141
No signiicant similarity; high identity to Teratosphaeria nubilosa
5538 – 5539
28S nrDNA
580
No signiicant similarity; high identity to Teratosphaeria nubilosa
22
PhylogenetiC lineages in the Capnodiales
Table 3. (Continued).
Isolate
Insertion
between
18S or 28S
nrDNA
Intron size
(bp)
Blast result
Teratosphaeria aff. nubilosa CBS 116283
4871 – 4872
28S nrDNA
141
No signiicant similarity; high identity to Teratosphaeria nubilosa
5538 – 5539
28S nrDNA
580
No signiicant similarity; high identity to Teratosphaeria nubilosa
1559 – 1560
18S nrDNA
403
52/61 of DQ471010 Rutstroemia irma 18S nrDNA
4875 – 4876
28S nrDNA
345
224/290 of EF115309 Cordyceps bassiana 28S nrDNA
5424 – 5425
28S nrDNA
478
47/50 of EF115313 Cordyceps bassiana 28S nrDNA
5538 – 5539
28S nrDNA
402
No signiicant similarity
1559 – 1560
18S nrDNA
403
52/61 of DQ471010 Rutstroemia irma 18S nrDNA
4875 – 4876
28S nrDNA
345
224/290 of EF115309 Cordyceps bassiana 28S nrDNA
5424 – 5425
28S nrDNA
478
47/50 of EF115313 Cordyceps bassiana 28S nrDNA
5538 – 5539
28S nrDNA
402
No signiicant similarity
954 – 955
18S nrDNA
316
129/158 of DQ518980 Lipomyces spencermartinsiae 26S
nrDNA
1559 – 1560
18S nrDNA
360
No signiicant similarity
1820 – 1821
18S nrDNA
388
128/168 of AF281670 Cryptendoxyla hypophloia 18S nrDNA
Teratosphaeria juvenalis CBS 110906
Teratosphaeria juvenalis CBS 111149
Teratosphaeria mexicana CBS 110502
Teratosphaeria mexicana CBS 120744
Teratosphaeria nubilosa CBS 115669
Teratosphaeria nubilosa CBS 116005
Teratosphaeria ohnowa CBS 112896
Teratosphaeria ohnowa CBS 112973
Teratosphaeria pseudosuberosa CBS 118911
Teratosphaeria sp. CBS 208.94
Teratosphaeria suberosa CPC 11032
Thedgonia-like genus CPC 12304
www.studiesinmycology.org
3560 – 3561
28S nrDNA
383
124/151 of EF647754 Thecaphora thlaspeos 28S nrDNA
4875 – 4876
28S nrDNA
327
99/114 of L81104 Gaeumannomyces graminis var. tritici 28S
nrDNA
5018 – 5019
28S nrDNA
315
No signiicant similarity
5424 – 5425
28S nrDNA
553
No signiicant similarity
954 – 955
18S nrDNA
318
130/158 of DQ518980 Lipomyces spencermartinsiae 26S
nrDNA
1559 – 1560
18S nrDNA
360
No signiicant similarity
1820 – 1821
18S nrDNA
389
85/109 of AF281670 Cryptendoxyla hypophloia 18S nrDNA
3560 – 3561
28S nrDNA
378
119/155 of AY298780 Lentinellus castoreus 18S nrDNA
4875 – 4876
28S nrDNA
327
162/200 of AB033530 Penicillium sabulosum 18S nrDNA
5018 – 5019
28S nrDNA
309
No signiicant similarity
5424 – 5425
28S nrDNA
659
No signiicant similarity
4871 – 4872
28S nrDNA
141
No signiicant similarity; high identity to Teratosphaeria aff.
nubilosa
5538 – 5539
28S nrDNA
580
No signiicant similarity; high identity to Teratosphaeria aff.
nubilosa
4871 – 4872
28S nrDNA
141
No signiicant similarity; high identity to Teratosphaeria aff.
nubilosa
5538 – 5539
28S nrDNA
580
No signiicant similarity; high identity to Teratosphaeria aff.
nubilosa
954 – 955
18S nrDNA
325
28/28 of DQ848329 Botryosphaeria quercuum 18S nrDNA
3560 – 3561
28S nrDNA
294
168/227 of FJ358267 Chaetothyriales sp. 28S nrDNA
5424 – 5425
28S nrDNA
607
47/48 of EF115313 Cordyceps bassiana 28S nrDNA
954 – 955
18S nrDNA
324
28/28 of DQ848329 Botryosphaeria quercuum 18S nrDNA
3560 – 3561
28S nrDNA
294
168/227 of FJ358267 Chaetothyriales sp. 28S nrDNA
5424 – 5425
28S nrDNA
607
47/48 of EF115313 Cordyceps bassiana 28S nrDNA
3560 – 3561
28S nrDNA
324
28/28 of DQ848329 Botryosphaeria quercuum 18S nrDNA
4875 – 4876
28S nrDNA
364
No signiicant similarity
954 – 955
18S nrDNA
342
No signiicant similarity
3560 – 3561
28S nrDNA
309
59/70 of AY207244 Mycena pura 28S nrDNA
4875 – 4876
28S nrDNA
296
44/51 of EF551317 Tremella globispora 28S nrDNA
5424 – 5425
28S nrDNA
313
159/197 of AB033529 Penicillium oblatum 18S nrDNA
5538 – 5539
28S nrDNA
596
80/99 of AB044639 Cordyceps kanzashiana 28S nrDNA
1820 – 1821
18S nrDNA
444
262/331 of EU167577 Mycosphaerella milleri 18S nrDNA
23
Dothidea insculpta DQ247802
Phaeotheca triangularis EU019279
Comminutispora agavaciensis Y18699
Incertae sedis
Phaeotheca fissurella EU981289
Racodium rupestre EU048576
Racodium rupestre EU048575
Racodium rupestre EU048577
Graphiopsis chlorocephala EU009458
Rachicladosporium luculiae EU040237
Rachicladosporium cboliae CPC 14034
Verrucocladosporium dirinae EU040244
Toxicocladosporium rubrigenum FJ790305
Toxicocladosporium irritans EU040243
Toxicocladosporium chlamydosporum FJ790301
Davidiella allicina CBS 723.79
Davidiellaceae
Davidiella macrospora DQ008148
Sphaerulina polyspora CBS 354.29
Cladosporium bruhnei EU019261
Cladosporium sp. FJ790289
Melanodothis caricis CBS 860.72
Cladosporium sp. CPC 15513
Cladosporium sp. CPC 15516
Cladosporium cladosporioides FJ890369
Scorias spongiosa DQ678075
Fumagospora capnodioides EU019269
Antennariella placitae GQ303299
Microxyphium theae GU301849
Polychaeton citri CBS 116435
Capnodiaceae
Leptoxyphium madagascariensis GQ303308
Leptoxyphium fumago CBS 123.26
Microxyphium citri AY004337
Capnodium coffeae DQ247800
Microxyphium aciculiforme GU301847
Condioxyphium gardeniorum GU301807
Devriesia strelitziicola CBS 122480
Anisomeridium consobrinum GU323215
Mycosphaerella eurypotami GU301852
Cystocoleus ebeneus EU048571
Cystocoleus ebeneus EU048573
Hortaea acidophila CBS 113389
Sporidesmium pachyanthicola DQ408557
Capnodiales sp. GU323217
Ramichloridium brasilianum EU041854
Capnodiales sp. GU323218
Staninwardia suttonii DQ923535
Capnodiales sp. GU323220
Teratosphaeria sp. GQ852712
Devriesia lagerstroemiae CPC 14403
Devriesia strelitziae EU436763
Teratosphaeria knoxdavesii EU707865
Capnodiales sp. GU323223
Tripospermum myrti GU323216
Passalora sp GQ852622
Devriesia hilliana CBS 123187
Penidiella strumelloidea EU019277
Capnobotryella renispora EU019248
Capnobotryella renispora CBS 215.90
Phacellium paspali GQ852627
Penidiella tasmaniensis DQ246233
Penidiella pseudotasmaniensis GQ852625
Capnodiales sp. GU323219
Teratosphaeria parva EU707875
Teratosphaeria jonkershoekensis GU301874
Catenulostroma protearum CPC 15370
Catenulostroma protearum CPC 15368
Catenulostroma protearum CPC 15369
Catenulostroma elginense EU019252
Capnodiales sp. GU323222
Capnodiales sp. GU323221
Xenomeris juniperi EF114709
Devriesia staurophora EF137359
Aulographina pinorum GU296138
Aulographina pinorum CBS 302.71
Catenulostroma microsporum EU167572
Catenulostroma abietis EU019249
Catenulostroma microsporum EU019255
Catenulostroma germanicum EU019253
Phaeothecoidea minutispora GQ852629
Bootstrap support values:
Phaeothecoidea
eucalypti EU019280
= 95 % and higher
Phaeothecoidea intermedia GQ852628
= 90 % to 94 %
Teratosphaeria mexicana DQ246237
= 80 % to 89 %
Teratosphaeria pseudosuberosa EU019256
Teratosphaeria suberosa CPC 11032
= 70 % to 79 %
Teratosphaeria suberosa DQ246235
Readeriella menaiensis GQ852670
Readeriella mirabilis GQ852662
0.1
Readeriella novaezelandiae DQ246239
Readeriella dendritica EU019271
Readeriella patrickii GQ852664
Readeriella tasmanica GQ852669
Readeriella eucalyptigena GQ852667
Readeriella dimorphospora EU019258
Readeriella angustia GQ852665
Readeriella nontingens EU019260
Readeriella minutispora EU019259
Readeriella callista GQ852655
Readeriella pseudocallista GQ852668
Readeriella eucalypti EU019289
Readeriellaa readeriellophora DQ246238
Teratosphaeriaceae
Crous et al.
fig. 1. RAxML tree using only the partial LSU alignment with bootstrap values after 1 000 pseudorepetitions on the nodes. Type strains and novel species described in this
study are indicated in bold.
24
PhylogenetiC lineages in the Capnodiales
Teratosphaeriaceae (continued)
Batcheloromyces proteae EU019247
Teratosphaeria sp. EU019307
Teratosphaeria ohnowa EU019305
Teratosphaeria secundaria EU019306
Teratosphaeria flexuosa FJ493216
Penidiella columbiana EU019274
Penidiella eucalypti EU882145
Penidiella tenuiramis GQ852626
Penidiella venezuelensis EU019278
Hortaea werneckii EU019270
Hortaea werneckii GU301818
Hortaea werneckii GU301817
Stenella araguata EU019250
Hortaea thailandica CPC 16651
Baudoinia compniacensis GQ852580
Piedraia quintanilhae CBS 327.63
Piedraia hortae var. hortae CBS 374.71
Piedraia hortae var. paraguayensis CBS 276.32
Piedraia hortae var. hortae CBS 375.71
Piedraia hortae var. hortae CBS 480.64
Piedraia hortae AY016366
Teratosphaeria cryptica CBS 110975
Teratosphaeria cryptica DQ246222
Teratosphaeria ovata FJ493218
Teratosphaeria sp. GQ852713
Teratosphaeria alboconidia FJ493221
Teratosphaeria brunneotingens EU019286
Teratosphaeria complicata GQ852714
Teratosphaeria miniata GQ852711
Teratosphaeria hortaea FJ790299
Teratosphaeria hortaea FJ790300
Teratosphaeria angophorae CBS 120493
Teratosphaeria verrucosa EU019293
Teratosphaeria juvenalis FJ493217
Teratosphaeria considenianae DQ923527
Teratosphaeria majorizuluensis GQ852710
Teratosphaeria corymbiae FJ493203
Teratosphaeria gauchensis EU019290
Teratosphaeria blakelyi DQ923526
Teratosphaeria zuluensis EU019296
Teratosphaeria stellenboschiana GQ852716
Teratosphaeria stellenboschiana EU019295
Teratosphaeria veloci FJ493223
Teratosphaeria suttonii EU019288
Teratosphaeria toledana DQ246230
Teratosphaeria nubilosa DQ246228
Teratosphaeria eucalypti DQ246225
Teratosphaeria viscidus FJ493204
Teratosphaeria destructans EU019287
Teratosphaeria sp. FJ493202
Teratosphaeria molleriana EU019292
Teratosphaeria profusa FJ493220
Teratosphaeria dimorpha FJ493215
Teratosphaeria macowanii EU019254
Teratosphaeria maxii DQ885899
Teratosphaeria maculiformis EU707867
Teratosphaeria proteae-arboreae EU707883
Teratosphaeria fibrillosa EU019282
Teratosphaeria fibrillosa GU323213
Ramichloridium apiculatum EU041848
Ramichloridium apiculatum EU041851
Dissoconium aciculare CBS 204.89
Dissoconium aciculare EU019266
Dissoconiaceae
Dissoconium australiensis GQ852588
Dissoconium dekkeri DQ204768
Dissoconium dekkeri CBS 111282
Dissoconium commune EU019267
Dissoconium commune CBS 110747
Zygophiala cryptogama FJ147157
Schizothyrium pomi EF134947
Schizothyriaceae
Zygophiala sp. FJ147159
Ramichloridium pini EU041859
Mycosphaerella parkii GQ852616
Pseudocercosporella sp. FJ031995
Mycosphaerella madeirae DQ204756
Polythrincium trifolii EU167612
Polythrincium trifolii EU167613
Polythrincium trifolii EU167610
Passalora vaginae GQ852624
Zasmidium aerohyalinosporum GQ852736
Mycosphaerella rosigena CBS 330.51
Mycosphaerella intermedia DQ246248
Mycosphaerella marksii GQ852612
Zasmidium nabiacense GQ852734
Phaeophleospora concentrica FJ493205
Brunneosphaerella protearum CPC 13905
Brunneosphaerella protearum CPC 16338
Brunneosphaerella protearum CPC 13914
Brunneosphaerella protearum CPC 15231
Periconiella arcuata EU041836
Mycosphaerellaceae
Verrucisporota daviesiae GQ852730
Periconiella velutina EU041838
Rasutoria tsugae EF114705
Rasutoria pseudotsugae EF114704
Verrucisporota proteacearum GQ852731
Penidiella nectandrae EU019275
Ramichloridium biverticillatum EU041853
Ramichloridium musae EU041857
Ramichloridium australiense EU041852
Ramichloridium strelitziae EU041860
Zasmidium citri GQ852733
Zasmidium anthuriicola GQ852732
Ramichloridium cerophilum EU041855
Zasmidium cellare EF137362
Zasmidium nocoxi GQ852735
Mycosphaerella aleuritidis EU167594
Bootstrap support values:
= 95 % and higher
= 90 % to 94 %
= 80 % to 89 %
= 70 % to 79 %
0.1
fig. 1. (Continued).
www.studiesinmycology.org
25
Bootstrap support values:
= 95 % and higher
= 90 % to 94 %
= 80 % to 89 %
= 70 % to 79 %
0.1
fig. 1. (Continued).
26
Ramularia nagornyi EU019257
Ramularia aplospora EU040238
Ramularia endophylla EU167569
Ramularia miae DQ885902
Ramularia pratensis var. pratensis EU019284
Ramularia brunnea EU167605
Mycosphaerella graminicola DQ678084
Mycosphaerella graminicola CBS 115943
Phaeophleospora eugeniicola FJ493209
Lecanosticta acicola GQ852598
Phaeophleospora eugeniae FJ493206
Mycosphaerella endophytica DQ246255
Mycosphaerella endophytica GQ852603
Mycosphaerella gregaria EU167580
Mycosphaerella pseudoendophytica DQ246253
Mycosphaerella stromatosa EU167598
Passalora daleae EU040236
Passalora sp. GQ852623
Passalora ageratinae CPC 15365
Passalora fulva DQ008163
Dothistroma septosporum GU350739
Dothistroma septosporum GQ852597
Dothistroma septosporum GU301853
Dothistroma pini GQ852596
Passalora bellynckii GQ852618
Mycosphaerella microsora EU167599
Mycosphaerella keniensis GQ852610
Passalora brachycarpa GQ852619
Mycosphaerella africana GQ852601
Mycosphaerella aurantia DQ246256
Mycosphaerella ellipsoidea GQ852602
Passalora graminis GQ852621
Passalora dalbergiae CPC 15419
Cercosporella virgaureae GQ852585
Ramulispora sorghi GQ852653
Cercospora beticola DQ678091
Cercospora zebrinae GQ852584
Cercospora apii GQ852583
Mycosphaerella coacervata EU167596
Mycosphaerella linorum EU167590
Septoria cucubali GQ852676
Septoria rubi EU167589
Septoria senecionis GQ852678
Septoria convolvuli GQ852675
Septoria apiicola GQ852674
Septoria leucanthemi GQ852677
Septoria populicola EU167578
Septoria aceris GQ852673
Mycosphaerella latebrosa CBS 687.94
Mycosphaerella flageoletiana EU167597
Mycosphaerella harthensis EU167602
Septoria berberidis EU167603
Mycosphaerella ribis EU167588
Sonderhenia eucalypticola DQ267574
Sonderhenia eucalyptorum DQ923536
Phaeophleospora atkinsonii CBS 124565
Phaeophleospora atkinsonii CBS 124566
Mycosphaerella colombiensis DQ204745
Phaeocryptopus gaeumannii EF114698
Mycosphaerella irregulariramosa GQ852609
Mycosphaerella acaciigena GQ852599
Mycosphaerella holualoana GQ852608
Mycosphaerella crystallina DQ204746
Mycosphaerella heimii GQ852604
Mycosphaerella heimioides GQ852607
Mycosphaerella konae GQ852611
Passalora eucalypti GQ852620
Pseudocercospora vitis DQ073923
Pseudocercospora sphaerulinae GQ852652
Mycosphaerella bixae GQ852630
Pseudocercospora gracilis DQ204750
Pseudocercospora robusta DQ204767
Pseudocercospora eucalyptorum DQ204762
Pseudocercospora pseudoeucalyptorum GQ852636
Pseudocercospora basitruncata DQ204759
Pseudocercospora platani GQ852635
Pseudocercospora fori DQ204748
Pseudocercospora crousii GQ852631
Pseudocercospora natalensis DQ267576
Pseudocercospora punctata GQ852645
Mycosphaerella milleri EU167577
Mycosphaerella pyri GQ852617
Pseudocercospora griseola f. griseola GU348997
Pseudocercospora griseola f. griseola GQ852633
Pseudocercospora fijiensis GQ852632
Pseudocercospora schizolobii GQ852646
Pseudocercospora tereticornis GQ852649
Pseudocercospora paraguayensis GQ852634
Pseudocercospora basiramifera DQ204761
Pseudocercospora sp. GQ852651
Mycosphaerellaceae
(continued)
Crous et al.
PhylogenetiC lineages in the Capnodiales
Magnaporthe grisea AB026819
Stomiopeltis betulae CBS 114420
Neofusicoccum australe CPC 10899
Graphiopsis chlorocephala CPC 11969
Toxicocladosporium irritans CBS 185.58
Davidiella tassiana CBS 723.79
Cladosporium bruhnei CBS 115683
Cladosporium bruhnei CBS 188.54
Cladosporium cladosporioides CBS 401.80
Cladosporium uredinicola ATCC 46649
Cladosporium cladosporioides CBS 109.21
Scorias spongiosa CBS 325.33
Leptoxyphium fumago CBS 123.26
Capnodium coffeae CBS 147.52
Pseudocercosporella fraxini CPC 11509
Mycosphaerella tasmaniensis CBS 111687
Phacellium paspali CBS 113093
Pseudocercospora-like genus CPC 10712
Capnobotryella renispora CBS 215.90
Capnobotryella renispora CBS 214.90
Pseudotaeniolina globosa CBS 109889
Devriesia staurophora CBS 375.81
Catenulostroma microsporum CBS 110890
Catenulostroma germanicum CBS 539.88
Readeriella mirabilis CBS 116293
Readeriella dimorphospora CBS 120034
Teratosphaeria mexicana CBS 110502
Teratosphaeria mexicana CPC 12349
Teratosphaeria suberosa CPC 11032
Teratosphaeria pseudosuberosa CBS 118911
Stenella araguata CBS 105.75
Teratosphaeria bellula CBS 111700
Catenulostroma chromoblastomycosum CBS 597.97
Catenulostroma elginense CBS 111030
Batcheloromyces proteae CBS 110696
Batcheloromyces leucadendri CPC 1837
Penidiella columbiana CBS 486.80
Teratosphaeria secundaria CBS 115608
Teratosphaeria sp. CBS 208.94
Teratosphaeria ohnowa CBS 112896
Teratosphaeria ohnowa CBS 112973
Teratosphaeria cryptica CBS 110975
Teratosphaeria stellenboschiana CBS 116428
Teratosphaeria fibrillosa CPC 1876
Teratosphaeria macowanii CBS 110756
Teratosphaeria macowanii CBS 111029
Teratosphaeria molleriana CPC 4577
Teratosphaeria molleriana CBS 111164
Teratosphaeria molleriana CBS 116370
Teratosphaeria alcornii CBS 313.76
Teratosphaeria verrucosa CPC 18
Teratosphaeria juvenalis CBS 110906
Bootstrap support values:
Teratosphaeria juvenalis CBS 111149
= 95 % and higher
Teratosphaeria nubilosa CBS 116005
= 90 % to 94 %
= 80 % to 89 %
Teratosphaeria nubilosa CBS 115669
= 70 % to 79 %
Teratosphaeria aff. nubilosa CBS 114419
Teratosphaeria aff. nubilosa CBS 116283
Teratosphaeria destructans CBS 111370
0.1
Teratosphaeria destructans CBS 111369
Teratosphaeria suttonii CPC 11279
Teratosphaeria suttonii CPC 12352
Teratosphaeria toledana CBS 113313
Teratosphaeria toledana CBS 115513
Davidiellaceae
Capnodiaceae
Teratosphaeriaceae
fig. 2. RAxML tree using the SSU, 5.8S nrDNA and LSU alignment with bootstrap values after 500 pseudorepetitions on the nodes.
www.studiesinmycology.org
27
Crous et al.
Bootstrap support values:
= 95 % and higher
= 90 % to 94 %
= 80 % to 89 %
= 70 % to 79 %
0.1
fig. 2. (Continued).
28
Staninwardia suttonii CBS 120061 Incertae
Passalora-like genus CPC 11876
Schizothyrium pomi CBS 228.57
Schizothyrium pomi CBS 486.50
Schizothyrium pomi CBS 406.61
Ramichloridium apiculatum CPC 12310
Dissoconium aciculare CBS 342.82
Dissoconium aciculare CBS 204.89
Dissoconium aciculare CBS 201.89
Dissoconium commune CBS 114238
Dissoconium commune CBS 110747
Dissoconium commune CBS 114239
Dissoconium dekkeri CBS 567.89
Dissoconium dekkeri CBS 111272
Dissoconium dekkeri CBS 110748
Dissoconium dekkeri CBS 111169
Dissoconium dekkeri CBS 111282
Passalora zambiae CBS 112971
Passalora zambiae CBS 112970
Passalora vaginae CBS 140.34
Ramichloridium-like genus CPC 10672
Mycosphaerella parkii CBS 387.92
Mycosphaerella marksii CBS 110942
Mycosphaerella marksii CPC 11222
Lecanosticta acicola CBS 871.95
Phaeophleospora eugeniicola CPC 2557
Phaeophleospora eugeniicola CPC 2558
Mycosphaerella stromatosa CBS 101953
Mycosphaerella endophytica CBS 114662
Mycosphaerella sp. CBS 111166
Mycosphaerella sp. CBS 111167
Mycosphaerella marasasii CBS 110790
Verrucisporota daviesiae CBS 116002
Verrucisporota proteacearum CBS 116003
Ramichloridium musae CBS 190.63
Ramichloridium cerophilum CBS 103.59
Zasmidium citri CBS 116366
Zasmidium anthuriicola CBS 118742
Thedgonia-like genus CPC 12304
Passalora sp. CPC 12319
Mycosphaerella lupini CPC 1661
Septoria-like genus CBS 102377
Passalora dodonaeae CPC 1223
Phloeospora maculans CBS 115123
Passalora perplexa CBS 116364
Passalora sequoiae CPC 11258
Passalora fulva CBS 119.46
Pseudocercospora opuntiae CBS 117708
Dothistroma septosporum CBS 112498
Dothistroma pini CBS 116487
Passalora bellynckii CBS 150.49
Mycosphaerella keniensis CBS 111001
Passalora sp. CPC 3951
Passalora brachycarpa CBS 115124
Mycosphaerella africana CBS 116154
Mycosphaerella ellipsoidea CBS 110843
Mycosphaerella graminicola CBS 100335
Ramularia rufomaculans CPC 10852
Mycosphaerella graminicola CBS 115943
Mycosphaerella graminicola CBS 110744
Ramularia sp. CBS 324.87
Ramularia endophylla CBS 113265
Ramularia grevilleana CPC 656
Ramularia sp. CPC 11297
Ramularia nagornyi CBS 120253
Ramularia acroptili CBS 120252
Ramularia brunnea CPC 4903
Ramularia pratensis var. pratensis CPC 11294
Ramularia sp. CPC 10066
Ramularia coleosporii CPC 11516
Ramularia uredinicola CPC 10813
sedis
Schizothyriaceae
Dissoconiaceae
Mycosphaerellaceae
PhylogenetiC lineages in the Capnodiales
Bootstrap support values:
= 95 % and higher
= 90 % to 94 %
= 80 % to 89 %
= 70 % to 79 %
0.1
Mycosphaerella handelii CBS 113302
Septoria quercicola CBS 663.94
Septoria lactucae CBS 352.58
Passalora eucalypti CBS 111318
Pseudocercospora angolensis CBS 149.53
Pseudocercospora angolensis CBS 112933
Pseudocercospora sphaerulinae CBS 112621
Pseudocercospora platani CBS 110755
Stigmina synanamorph CPC 11721
Pseudocercospora ocimicola CPC 10283
Pseudocercospora punctata CPC 10532
Pseudocercospora punctata CBS 113315
Mycosphaerella pyri CBS 222.31
Pseudocercospora griseola f. griseola CBS 194.47
Pseudocercospora griseola f. griseola CBS 880.72
Mycosphaerella bixae CBS 111804
Pseudocercospora luzardii CPC 2556
Pseudocercospora humuli CPC 11358
Pseudocercospora protearum var. leucadendri CPC 1869
Pseudocercospora pseudoeucalyptorum CBS 114242
Pseudocercospora kaki CPC 10636
Pseudocercospora cruenta CBS 462.75
Pseudocercospora vitis CPC 11595
Pseudocercospora paraguayensis CBS 111317
Pseudocercospora fijiensis X300
Pseudocercospora atromarginalis CPC 11372
Pseudocercospora pallida CPC 10776
Pseudocercospora fuligena CPC 12296
Pseudocercospora cordiana CBS 114685
Pseudocercospora cruenta CPC 10846
Pseudocercospora macrospora CBS 114696
Pseudocercospora chengtuensis CPC 10785
Passalora graminis CBS 113303
Sonderhenia eucalypticola CPC 11252
Mycosphaerella irregulariramosa CBS 111211
Mycosphaerella heimioides CBS 111190
Mycosphaerella heimii CBS 110682
Mycosphaerella holualoana CBS 110699
Mycosphaerella acaciigena CBS 112516
Mycosphaerella acaciigena CBS 112515
Cercosporella virgaureae CBS 113304
Ramulispora sorghi CBS 110579
Ramulispora sorghi CBS 110578
Pseudocercosporella capsellae CPC 10301
Pseudocercospora sp. CPC 11592
Mycosphaerella latebrosa CBS 687.94
Mycosphaerella latebrosa CBS 652.85
Pseudocercosporella sp. CPC 11414
Miuraea persicae CPC 10069
Septoria rosae CBS 355.58
Passalora dioscoreae CPC 10855
Pseudocercospora eucommiae CPC 10802
Cercospora janseana CBS 145.37
Cercospora beticola CBS 116456
Cercospora zebrinae CBS 118790
Cercospora zebrinae CBS 118789
Cercospora sojina CPC 12322
Cercospora zebrinae CBS 112893
Cercospora capsici CPC 12307
Cercospora apii CBS 118712
Pseudocercosporella sp. CPC 10050
Septoria cucubali CBS 102368
Septoria protearum CPC 1470
Pseudocercosporella sp. CPC 4008
Pseudocercosporella sp. CBS 112737
Septoria leucanthemi CBS 109090
Septoria apiicola CBS 400.54
Septoria obesa CBS 354.58
Septoria convolvuli CBS 102325
Septoria senecionis CBS 102366
Septoria dysentericae CPC 12328
Mycosphaerellaceae
(continued)
fig. 2. (Continued).
www.studiesinmycology.org
29
Crous et al.
reSulTS
DNA ampliication and phylogeny
Ampliication products of approximately 1 700 bases were obtained
for the standard ampliication of the isolates listed in Table 1. The
LSU region of these sequences was used to obtain additional
sequences from GenBank that were added to the partial LSU
alignment. We expected a total size of approximately 5 500 bp for
the concatenated SSU, ITS1, 5.8S nrDNA, ITS2 and LSU at the start
of the study; however, our alignment totalled about 12 000 bp due
to numerous insertions (most likely group 1 introns) encountered
for several strains (Table 3). These insertions frequently resulted
in products too large to amplify or sequence effectively and
sometimes required us to design additional novel primers in extra
overlapping steps to complete these gaps (see Materials and
Methods for details). Searching the GenBank database using these
insertions had varied success (Table 3). Sequences of the 18S
nrDNA are more abundant in the database whereas sequences of
the second half to two-thirds of the 28S nrDNA are mostly absent.
This also evident in Table 3, where insertions in the SSU more
frequently found with similarity sequences in the database and
insertions in the LSU (e.g. those between positions 5018–5019 and
5424–5425) frequently did not retrieve any signiicant similarity.
Although there were some exceptions (e.g. the insertion between
1820 and 1821 in the SSU of Batcheloromyces leucadendri), most
of the insertions in the SSU obtained hits with SSU sequences of
species of Capnodiales in the database. In one case, between
954 and 955 for the SSU sequence of Teratosphaeria mexicana
(both strains), a partial hit was obtained with an LSU sequence
of Lipomyces spencermartinsiae (GenBank DQ518980). Many of
the insertions in the LSU sequences did not retrieve signiicant hits
in the database and those that did were with unrelated taxa. It is
quite possible that this is an artifact of the poor representation of
full-length LSU sequences in the database, especially for members
of the Capnodiales. In some cases, an LSU insertion retrieved
a hit with SSU sequences in the database, e.g. the insertion
between 5538 and 5539 in Batcheloromyces proteae and between
3560 and 3561 and 4875 and 4876 in Teratosphaeria mexicana
strain CBS 120744. In two cases (Mycosphaerella latebrosa and
Phaeophleospora eugeniicola), an insertion was either lost or gained
between two strains of the same species. The primers designed in
this study allowed us to effectively amplify and sequence the SSU
and LSU for the selected isolates. Althought these primers were
not tested on taxa outside of the Capnodiales (except for one of the
outgroups, Neofusicoccum australe), we attempted to design them
as robust as possible using degeneracy if needed. We therefore
expect that these primers will have wider applicability than just the
Capnodiales in cases where other published primers fail to amplify
or amplify poorly.
The RAxML search of the partial LSU alignment yielded a
most likely tree (Fig. 1) with a log likelihood -13397.994021. The
matrix had 395 distinct alignment patterns, with 6 % completely
undetermined characters in the alignment. The manually adjusted
alignment contained 295 sequences (including the outgroup
sequence, Dothidea insculpta GenBank DQ247802) and 763
characters including alignment gaps. The RAxML search of the
almost complete SSU, 5.8S nrDNA and LSU alignment yielded a
most likely tree (Fig. 2) with a log likelihood -39022.881140. The
matrix had 1211 alignment patterns with 0.01 % of the characters
consisting of gaps or undetermined characters. The manually
adjusted alignment contained 205 sequences (including the
30
outgroup sequences, Neofusicoccum australe CPC 10899 and
Magnaporthe grisea GenBank AB026819) and 5110 characters
including alignment gaps. The obtained phylogenies (Figs 1–2) are
discussed in the Taxonomy section below.
Taxonomy
Several well-supported clades could be distinguished in the present
study (Figs 1–2), correlating to families in the Capnodiales. These
families, and several new genera and species, are treated below.
Treatment of phylogenetic clades
Capnodiales Woron. Ann. Mycol. 23: 177. 1925.
Data obtained from multi-gene phylogenies prompted Schoch
et al. (2006) to merge Mycosphaerellales with Capnodiales.
Although the present study included numerous additional isolates,
the orders remain problematic. Although there is support for the
Mycosphaerellales as an order, additional families such as the
Schizothyriaceae and Dissoconiaceae (see below) would have
to also be elevated to order level, which would result in orders
containing a single family, while Teratosphaeriaceae appears to
comprise unresolved lineages. For this reason it was decided to
retain these families within Capnodiales, but noting that as more
families are added and better circumscribed, it is quite possible that
the Mycosphaerellales would again be resurrected.
Mycosphaerellaceae Lindau, In: Engler & Prantl, Nat. Planzenfamilien 1(1): 421. 1897.
Type species: Mycosphaerella punctiformis (Pers. : Fr.) Starbäck,
Bih. Kongl. Svenska Vetensk.-Akad. Handl. 15(3, 2): 9. 1889.
Notes: The Mycosphaerellaceae contains numerous genera,
20 of which are listed by Crous (2009), with many names
under consideration (Crous et al. 2009b, c). From these data
it is clear that genera such as Passalora, Pseudocercospora,
Pseudocercosporella, Septoria, Zasmidium and Ramichloridium
are paraphyletic (Hunter et al. in prep.). Well-resolved genera
include Cercospora, Cercosporella, Ramularia, Ramulispora,
Sonderhenia and Polythrincium. One particularly problematic
clade contains Periconiella, Ramichloridium, Verrucisporota
and Zasmidium, along with “Mycosphaerella” and Rasutoria
teleomorphs. Barr (1987) erected Rasutoria for species with brown
ascospores occurring on Gymnospermae. Rasutoria clusters
in a clade adjacent to “Mycosphaerella” species with hyaline
ascospores, such as M. aleuritidis and Mycosphaerella daviesiicola
(Verrucisporota daviesiae) (Beilharz & Pascoe 2002).
The genus Phaeophleospora (1916) clusters with
Lecanosticta acicola. The genus Lecanosticta (1922) has typical
Phaeophleospora-like conidia, except that its conidiomata are
acervular, and not pycnidial. If the type of Lecanosticta, L. pini
also clusters in this clade, the generic concept Phaeophleospora
may have to be widened to include Lecanosticta, as was done with
Kirramyces to include Colletogloeopsis (Cortinas et al. 2006a, b).
Considerable controversy has surrounded the coelomycetes
that Crous et al. (1997) placed in Phaeophleospora. Based on
DNA phylogenetic data, it has now been shown that Kirramyces
anamorphs (Walker et al. 1992), including those accommodated
in Colletogloeopsis (Crous & Wingield 1996, Crous et al. 2004c,
2006c, Cortinas et al. 2006a, b), are linked to Teratosphaeria
(Andjic et al. 2007, Crous et al. 2009b, c). Crous et al. (2007a)
PhylogenetiC lineages in the Capnodiales
showed Phaeophleospora to reside in the Mycosphaerellaceae and
Kirramyces in the Teratosphaeriaceae, respectively. However, most
taxa investigated to date were collected from Eucalyptus. As shown
in the present study, Phaeophleospora atkinsonii, a pathogen of
Hebes spp. (Wu et al. 1996, Pennycook & McKenzie 2002), clusters
distant from Phaeophleospora s. str., while the same is true for
Phaeophleospora concentrica, which is a pathogen of Protea spp.
(Taylor et al. 2001a), and Phaeophleospora stonei, a pathogen of
Eucalyptus (Crous et al. 2007c, 2009c). These taxa thus clearly
represent yet another two genera in the Phaeophleospora complex.
An older name that would potentially be available is Scoleciasis.
However, when B. Sutton examined exsiccati of the type species,
S. aquatica, only ascomata of a Leptosphaeria species were
found (Crous et al. 1997). The association of S. aquatica with the
Leptosphaeria was also noted in the original description, and this may
indicate that Scoleciasis is allied to taxa in the Phaeosphaeriopsis/
Phaeoseptoria complex (Arzanlou & Crous 2006). Both P. atkinsonii
and P. concentrica have a typical Kirramyces morphology, namely
brown, percurrently proliferating conidiogenous cells, and brown,
obclavate, verruculose, transversely euseptate conidia. Further
species thus need to be included in analyses before these generic
concepts can be clariied.
During the course of this study several fresh collections of
Leptosphaeria protearum were obtained. Leptosphaeria protearum
is a major leaf spot and blight pathogen of Protea spp. (KnoxDavies et al. 1987), and causes severe losses in plantations of
South African Protea spp. in Hawaii, and has been recorded in
many countries where South African proteas are cultivated (Taylor
& Crous 1998, Taylor et al. 2001b, Crous et al. 2004a). Cultures
of this pathogen were found to cluster in the Mycosphaerellaceae,
where they represent an undescribed genus, characterised
by having bitunicate asci without pseudoparaphyses, brown,
3-septate ascospores, and a Coniothyrium-like anamorph. Its close
phylogenetic relationship to Phaeophloeospora concentrica (Fig. 1)
suggests that they could be congeneric, and that in future more
Phaeophloeospora-like anamorphs may be found to cluster in this
clade. We propose a new genus to accommodate Leptosphaeria
protearum below.
Brunneosphaerella Crous, gen. nov. MycoBank MB514694.
Etymology: Brunneus + Sphaerella = is after its brown ascospores
and Sphaerella-like morphology.
Mycosphaerellae similis, sed ascosporis brunneis, 3-septatis.
Ascomata amphigenous, immersed to semi-immersed, black,
single, gregarious, substomatal, pyriform or globose with a papillate,
periphysate ostiole. Peridium consisting of three strata of slightly
compressed textura angularis, an outer stratum of dark brown, thickwalled cells, becoming paler in the central stratum, and hyaline,
thin-walled in the inner stratum. Asci clavate to cylindro-clavate,
often curved, tapering to a pedicel, narrowing slightly to a rounded
apex with an indistinct ocular chamber, 8-spored, bitunicate with
issitunicate dehiscense. Pseudoparaphyses absent. Ascospores
biseriate, fusiform, broader at the apical end, initially hyaline and
1-septate, becoming yellow-brown and 3-septate at maturity,
slightly constricted at median to supra-median septum.
Type species: Brunneosphaerella protearum (Syd. & P. Syd.) Crous,
comb. nov.
www.studiesinmycology.org
Brunneosphaerella jonkershoekensis (Marinc., M.J.
Wingf. & Crous) Crous, comb. nov. MycoBank MB514695.
Fig. 3.
Basionym: Leptosphaeria jonkershoekensis Marinc., M.J. Wingf. &
Crous, In: Marincowitz et al., Microfungi occurring on Proteaceae in
the fynbos: 62. 2008.
Ascomata pseudothecial, subepidermal, immersed, obpyriform,
papillate, 180–205 × 160–235 µm. Peridium 20–30 µm thick,
composed of relatively large cells, 11–15 × 2.5–5.5 µm; cells
arranged in three strata; outer stratum consisting of 3–5 layers of dark
brown, very thick-walled cells; middle stratum transient, consisting
of a few layers of pale brown, thick-walled, compressed cells; inner
stratum consisting of 1–2 layers of thin-walled, very compressed
cells. Pseudoparaphyses absent. Asci bitunicate, inlated cylindrical
to clavate, 81–95 × 13–15 µm, ocular chamber dome-shaped,
indistinct. Ascospores pale brown, fusoid to ellipsoidal, tapering
towards the base, (25–)29–34(–36) × (5–)6–7(–9) µm (av. 31.4 ×
6.7 µm), apical cell the shortest, upper hemispore slightly larger
than lower, at times slightly curved, 3-septate, smooth, guttulate
(adapted from Marincowitz et al. 2008).
Host range and geographic distribution: Protea repens (South
Africa, Western Cape) (Marincowitz et al. 2008).
Specimen examined: South Africa, Western Cape Province, Jonkershoek Nature
Reserve, leaf litter of Protea repens, 6 Jun. 2000, S. Marincowitz, PREM 59447
holotype.
Notes: Although no culture is presently available for this species, it
clearly represents a species of Brunneosphaerella, characterised
by its bitunicate asci, and brown, 3-septate ascospores, as
well as the absence of pseudoparaphyses. Brunneosphaerella
jonkershoekensis can easily be distinguished from B. protearum
based on its much larger ascospores (Crous et al. 2004a).
Brunneosphaerella protearum (Syd. & P. Syd.) Crous,
comb. nov. MycoBank MB514696. Fig. 4.
Basionym: Leptosphaeria protearum Syd. & P. Syd., Ann. Mycol.
10: 441. 1912.
Anamorph: “Coniothyrium” protearum Joanne E. Taylor & Crous,
IMI Descriptions of Fungi and Bacteria No. 1343. 1998.
Leaf spots circular to irregular, discrete to conluent, variable in size,
under conditions favourable to disease symptoms more similar
to a blight than a leaf spot, necrotic, sunken with a raised dark
brown margin and with conspicuous black ascomata in the dead
tissue, 4–30 mm diam. Ascomata pseudothecial, substomatal,
amphigenous, immersed to semi-immersed, not erumpent, black,
single, gregarious, 180–320 µm diam; in section, substomatal,
subepidermal, pyriform or globose with a papillate, periphysate
ostiole, immersed in a stroma consisting of deteriorated host
mesophyll cells illed with fungal hyphae, (210–)230–264(–288) µm
high, (180–)200–255(–300) µm diam. Peridium consisting of three
strata of slightly compressed textura angularis, an outer stratum of
dark brown, thick-walled cells, becoming paler in the central stratum,
and hyaline, thin-walled in the inner stratum, altogether (20–)24.5–
37.5(–50) µm thick. Asci clavate to cylindro-clavate, often curved,
tapering to a pedicel, narrowing slightly to a rounded apex with an
indistinct ocular chamber, 8-spored, bitunicate with issitunicate
dehiscense, (70–)80–87.5(–105) × (13.5–)14.5–16(–21.5) µm.
Pseudoparaphyses absent. Ascospores biseriate, fusiform, broader
31
Crous et al.
fig. 3. Brunneosphaerella jonkershoekensis. A–B. Vertical sections through ascomata showing wall structure. C–D, G. Bitunicate asci. E–F. Ascospores. Scale bars: A, C = 50
µm, B = 20 µm, D, G = 10 µm, E–F = 5 µm (from Marincowitz et al. 2008).
at the apical end, initially hyaline and 1-septate, becoming yellowbrown and 3-septate at maturity, slightly constricted at median to
supra-median septum, (21.5–)27.5–29.5(–37.5) × (6.3–)7.5–8(–10)
µm in water mounts, (21–)25.5–27.5(–31) × (5.5–)6–7(–8) µm in
lactophenol. Conidiomata barely visible and interspersed between
ascomata, pycnidial, subepidermal, substomatal, separate,
globose to pyriform, occasionally with well-developed papilla, dark
brown, < 200 µm diam. Conidiophores reduced to conidiogenous
cells. Conidiogenous cells discrete, smooth, hyaline, doliiform to
ampulliform, holoblastic, proliferating 1–2 times percurrently, 4–6
× 3–4 µm. Conidia pale brown to medium brown, thick-walled on
maturity, smooth to inely verruculose, eguttulate, ellipsoidal to
globose, often truncate at one end, 5–10 × 3–7 µm (adapted from
Crous et al. 2004a).
Host range and geographic distribution: Protea cynaroides, P.
‘Susara’ (Portugal, Madeira) (Moura & Rodrigues 2001); P. caffra,
P. compacta, P. cynaroides, P. gaguedi, P. grandiceps, P. lacticolor,
P. laurifolia, P. lepidocarpodendron, P. lorifolia, P. magniica, P.
nitida, P. punctata, P. repens, P. ‘Sheila’, Protea spp. (South Africa);
P. cynaroides, P. laurifolia, P. neriifolia, P. ‘Ivory Musk’, P. ‘Mink’, P.
‘Pink Ice’, P. ‘Rose Mink’, P. susannae, Protea sp. (U.S.A., Hawaii)
(Taylor et al. 2001b); P. cynaroides, P. gaguedi, P. neriifolia, Protea
sp. (Zimbabwe, Inyanga) (Masuka et al. 1998).
Specimens examined: South Africa, Western Cape Province, Bettys’ Bay, leaf
litter of Protea magniica, 11 Jul. 2000, S. Marincowitz, PREM 59448; Helderberg
Nature Reserve, leaf litter of Protea laurifolia, 14 Aug. 2000, S. Marincowitz, PREM
59482; Helderberg Nature Reserve, leaf litter of Protea obtusifolia, 14 Aug. 2000,
S. Marincowitz, PREM 59495; Jonkershoek Nature Reserve, leaf litter of Protea
32
nitida, 6 Jun. 2000, S. Marincowitz, PREM 59442; Jonkershoek Nature Reserve,
leaf litter of Protea repens, 6 Jun. 2000, S. Marincowitz, PREM 59450; Jonkershoek
Nature Reserve, S33°59’11.2” E18°57’14.7” leaves of Protea sp., 1 Apr. 2007, P.W.
Crous, CBS H-20330, cultures CPC 13914–13916; Jonkershoek Nature Reserve,
S33°59’26.1” E18°57’59.5” leaves of Protea repens, 1 Apr. 2007, P.W. Crous, CBS
H-20331, cultures CPC 13911–13913; Jonkershoek Nature Reserve, leaves of
Protea sp., 1 Apr. 2007, P.W. Crous, CBS H-20332, cultures CPC 13908–13910;
Jonkershoek Nature Reserve, “Tweede Waterval”, leaves of Protea sp., 1 Apr.
2007, P.W. Crous, CBS H-20333, cultures CPC 13902–13907; Jonkershoek Nature
Reserve, leaves of Protea nitida, 12 Apr. 2008, L. Mostert, CBS H-20334, cultures
CPC 15231–15233; Kirstenbosch Botanical Garden, leaves of Protea sp., 13 Jan.
2009, P.W. Crous, CBS H-20335, culture CPC 16338.
Notes: Although Taylor & Crous (1998) reported a Coniothyriumlike anamorph to develop in culture, none of the cultures examined
in the present study on MEA, PDA or OA could be induced to
sporulate, though spermatogonia and ascomatal initials were
commonly observed.
The fact that cultures of Leptosphaeria protearum, which
represents a well-known and serious pathogen of Proteaceae,
clustered in the Mycosphaerellaceae, was totally unexpected.
A further surprise was the fact that this species appears to
represent a complex of several cryptic taxa. Whether these taxa
can be correlated with differences in host range and geographic
distribution can only be resolved once more collections have
been obtained for study. Although the genus Sphaerulina, which
represents Mycosphaerella-like taxa with 3-septate, hyaline
ascospores, is part of the Mycosphaerellaceae (Crous et al.,
unpubl data), the type species, S. myriadea, clusters in the
Septoria clade, and is thus unavailable for the species occurring on
Proteaceae. Morphologically Brunneosphaerella is also distinct in
PhylogenetiC lineages in the Capnodiales
fig. 4. Brunneosphaerella protearum. A–D. Leaf spots on different Protea spp. E. Close up of leaf spot showing ascomata. F. Substomatal ascomata. G–H. Vertical sections
though ascomata, showing wall structure. I–K. Germinating ascospores on MEA. L–M, R. Bitunicate asci. N–Q, S. Juvenile to mature ascospores. Scale bars: G = 75 µm, H
= 10 µm.
www.studiesinmycology.org
33
Crous et al.
that ascospores are always brown at maturity, and anamorphs have
brown, percurrently proliferating conidiogenous cells, appearing
Phaeophleospora-like. The recognition of Brunneosphaerella
as a distinct genus in the Mycosphaerellaceae also raises the
intriguing possibility that many phytopathogenic species of the
Leptosphaeria-complex with brown, 3-septate ascospores, but
lacking paraphyses, actually belong to Brunneosphaerella.
Passalora ageratinae Crous & A.R. Wood, sp. nov. MycoBank MB514697. Fig. 5.
Culture characteristics: On MEA erumpent, with uneven, folded
surface, lobate margin, and moderate aerial mycelium; centre pale
mouse-grey with patches of cinnamon, outer margin olivaceousgrey; reverse olivaceous-grey with patches of cinnamon; reaching
15 mm diam; on PDA spreading, with cinnamon to cream patches
in centre, becoming umber towards smooth margins, with diffuse
red pigment in agar; reverse olivaceous-grey, with patches of
red, reaching 15 mm diam; on OA lat, spreading, up to 30 mm
diam, iron-grey, with white, solitary mycelia strands, though aerial
mycelium generally absent, reaching 30 mm diam.
Etymology: Named after the host on which it occurs, Ageratina
adenophora.
Host range and geographic distribution: Ageratina adenophora,
Australia, South Africa.
Passalorae assamensis similis, sed coloniis amphigenis, sine mycelio externo,
conidiophoris brevioribus, 15–40 × 3–4.5 µm.
Specimen examined: South Africa, KwaZulu-Natal Province, Hilton, on leaves of
Ageratina adenophora, 28 May 2008, A.R. Wood, CBS H-20336 holotype, cultures
ex-type CPC 15365 = CBS 125419, CPC 15366, 15367.
Leaf spots amphigenous, angular to irregular, 2–8 mm diam,
medium brown, frequently with pale to grey-brown central part,
and raised, dark brown border; pale to medium brown in reverse,
with raised, dark brown border. Mycelium internal, consisting of
smooth, branched, pale brown, 2–3 µm wide hyphae. Caespituli
fasciculate, amphigenous, medium brown, arising from a brown,
erumpent stroma, up to 80 µm wide, 40 µm high. Conidiophores
subcylindrical, straight to geniculous-sinuous, unbranched,
medium brown, inely verruculose, 1–3-septate, 15–40 × 3–4.5
µm. Conidiogenous cells terminal, pale to medium brown, inely
verruculose with terminal, sympodial conidiogenous loci that are
1–2 µm diam, slightly thickened, darkened and refractive, 10–20 ×
3–4 µm. Conidia in unbranched chains, pale brown, smooth, inely
to prominently guttulate, subcylindrical to narrowly obclavate, apex
obtuse, base long obconically subtruncate, (0–)1–3(–5)-septate,
(20–)30–60(–80) × (3–)4(–4.5) µm; hila 1–1.5 µm wide, somewhat
thickened, darkened and refractive.
Notes: Ageratina adenophora (crofton weed; Asteraceae), which
is indigenous to Mexico, has invaded many countries as a rapidly
growing weed, forming dense thickets (Morris 1989, Parsons &
Cuthbertson 1992, Wagner et al. 1999, Zhu et al. 2007, Muniappan
et al. 2009). It is considered a serious weed in agriculture and
forestry (Bess & Haramoto 1958, Sharma & Chhetri 1977, Kluge
1991), often replacing more-desired vegetation or native species.
A leaf spot pathogen, originally misidentiied as Cercospora
eupatorii (this species is currently known as Pseudocercospora
eupatorii), was found to infect plants in Australia where a stem
galling ly (Procecidochares utilis; Tephritidae) was introduced from
Hawaii as a biological control agent (Dodd 1961). Presumably the
fungus was introduced together with the lies originally from Mexico
to Hawaii and then to Australia. Subsequently this same fungus
was obtained from Australia and released in South Africa after
host speciicity testing indicated it was restricted to A. adenophora
fig. 5. Passalora ageratinae. A. Leaf spots. B. Close up of leaf spot with fruiting structures. C–D. Conidiophores. E–J. Conidia. Scale bars = 10 µm.
34
PhylogenetiC lineages in the Capnodiales
(Morris 1989). The fungus causes partial defoliation of mature
plants (Dodd 1961, Auld 1969), though the impact depends on
environmental conditions (Dodd 1961). Seedlings are however
killed rapidly (Wang et al. 1997).
This fungus, which has hitherto been known simply as
“Phaeoramularia” sp., still lacks a name and proper description.
The genus Phaeoramularia is treated as a synonym of Passalora
(Crous & Braun 2003), and hence the species is named in the latter
genus as P. ageratinae. Interestingly, this species appears to be
closely related to Passalora fulva, which is a serious pathogen of
tomato (Solanaceae) (Thomma et al. 2005).
Passalora armatae Crous & A.R. Wood, sp. nov. MycoBank
MB514698. Fig. 6.
Etymology: Named after the host on which it occurs, Dalbergia
armata.
Passaloraea dalbergiicolae similis, sed conidiophoris in synnematibus densis,
conidiis ad basim obconice truncatis, apice rostrato.
Leaf spots amphigenous, on upper surface visible as red-brown,
irregular to subcircular spots with indistinct margins, 0.5–2 mm
diam; in reverse indistinct, chlorotic to medium or red-brown.
Mycelium internal, consisting of smooth, branched, pale brown,
2–3 µm wide hyphae. Caespituli hypophyllous, fasciculate to
synnematous, up to 200 µm high and 250 µm wide, situated on
a prominently erumpent, pale brown stroma, up to 100 µm high
and wide. Conidiophores subcylindrical, unbranched, lexuous,
guttulate, pale to medium brown, smooth, 120–180 × 4–6 µm,
2–6-septate. Conidiogenous cells terminal, subcylindrical,
guttulate, pale to medium brown, inely verruculose, becoming
somewhat swollen, appearing slightly clavate, 25–70 × 6–8 µm;
conidiogenous loci 4–20 per conidiogenous cell, sympodial, round,
darkened, thickened, refractive, prominent, 2–3 µm wide, up to
1 µm high. Conidia (27–)30–40(–45) × 9–10(–12) µm, pale to
medium brown, smooth to inely verruculose, granular to guttulate,
thin-walled, ellipsoidal to obovoid, transversely 2–4-euseptate,
widest in middle of basal cell, or middle of conidium, tapering to an
obconically truncate base; hilum thickened, darkened and refractive;
apical cell conical, elongating to an apical beak up to 20 µm long.
When cultivated conidia remain attached to conidiogenous cells,
giving conidiophores the appearance of small tufts which is very
characteristic, and not commonly observed in Passalora.
Culture characteristics: On MEA slow growing, erumpent, with
dense white aerial mycelium, which becomes mouse-grey, reaching
5 mm diam after 1 wk; on PDA mouse-grey (surface), iron-grey
(reverse), with diffuse red pigment in agar; on OA similar to PDA,
also with diffuse red pigment in agar.
Host range and geographic distribution: Dalbergia armata, South
Africa.
Specimen examined: South Africa, KwaZulu-Natal Province, South Coast, Mpenjati
Nature Reserve, between Ramsgate and Port Edward, on leaves of Dalbergia
armata, 28 May 2008, A.R. Wood, CBS H-20337 holotype, cultures ex-type CPC
15419 = CBS 125420, CPC 15420, 15421.
Notes: Passalora dalbergiae, which occurs on Dalbergia sissoo
(Fabaceae) in India, is distinct from P. armatae in having supericial
mycelium and solitary conidiophores (Hernández-Gutiérrez &
fig. 6. Passalora armatae. A. Fruiting in vivo. B–C. Caespituli with prominent basal stroma. D. Sporulation on MEA. E. Conidiogenous cells giving rise to conidia. F–G. Conidia.
Scale bars: B = 125 µm, C–E = 10 µm.
www.studiesinmycology.org
35
Crous et al.
Dianese 2009). The previously described Passalora dalbergiicola
is similar to P. armatae in conidial dimensions (3-septate, 25–45 ×
7–10 µm; Ellis 1976), but distinct in that conidiophores are not in
dense synnemata, conidiogenous cells can have single apical loci,
and conidia have a less prominent basal taper, and lack the apical
beaks typical of P. armatae (in vivo and in vitro).
Schizothyriaceae Höhn. ex Trotter, Sacc., D. Sacc. & Traverso, In: Saccardo, Syll. Fung. 24(2): 1254. 1928.
Type species: Schizothyrium acerinum Desm., Ann. Sci. Nat. Bot.
11: 360. 1849.
Notes: Members of the Schizothyriaceae are associated with
lyspeck symptoms on apples and pear fruit. The fungi grow
supericially on the epicuticular wax, thereby reducing the
marketability of the fruit, but do not penetrate the cuticle (Belding
et al. 2000). Batzer et al. (2005, 2007) reported a range of diverse
fungi to be associated with lyspeck symptoms on apples, the most
prominent being species of Schizothyrium.
Dissoconiaceae Crous & de Hoog, fam. nov. MycoBank
MB514699.
Ascomata
pseudotheciales,
immerse,
globosa,
uniloculares.
Sine
pseudoparaphysibus. Asci fasciculati, octospori, bitunicati. Ascosporae ellipsoideaefusiformes, 1-septatae, hyalinae. Conidiophora separata, ex hyphis oriunda,
subcylindrica, subulata, lageniformia vel cylindrica, apicem versus attenuata, apice
obtuse rotundato vel truncate, recta vel semel geniculata, laevia, modice brunnea,
0–pluriseptata, locis terminalibus vel lateralibus, rhachidi cum cicatricibus leniter
incrassates, fuscatis. Conidia solitaria, pallide olivaceo-brunnea, laevia, ellipsoidea,
obclavata vel globosa, 0–1-septata, hilis aliquantum fuscatis. Conidia secundaria
nulla vel formata ad conidia primaria, pallide olivacea vel subhyalina, aseptata,
pyriformia; conidiis impigre vel passive emittentibus.
Ascomata pseudothecial, immersed, globose, unilocular, papillate,
ostiolate, canal periphysate; wall consisting of 3–4 layers of
brown textura angularis; inner layer of lattened, hyaline cells.
Pseudoparaphyses absent. Asci fasciculate, 8-spored, bitunicate.
Ascospores ellipsoid-fusoid, 1-septate, hyaline, with or without
mucoid sheath. Mycelium internal and external, consisting of
branched, septate, smooth, hyaline to pale brown hyphae.
Conidiophores separate, arising from hyphae, subcylindrical,
subulate or lageniform to cylindrical, tapering to a bluntly rounded
or truncate apex, straight to once geniculate, smooth, medium
brown, 0–multi-septate; loci terminal and lateral, visible as slightly
thickened, darkened scars on a rachis. Conidia solitary, pale
olivaceous-brown, smooth, ellipsoid to obclavate or globose,
0–1-septate; hila somewhat darkened. Secondary conidia present
or absent; developing adjacent to primary conidia, pale olivaceous
to subhyaline, aseptate, pyriform; conidium discharge active or
passive.
Type species: Dissoconium aciculare de Hoog, Oorschot &
Hijwegen, Proc. K. Ned. Akad. Wet., Ser. C, Biol. Med. Sci. 86(2):
198. 1983.
Notes: Species of Dissoconium have Mycosphaerella-like
teleomorphs (Crous et al. 2004c). The genus is characterised by
forming conidia in pairs that are forcefully discharged, which is
quite unique in the Capnodiales (de Hoog et al. 1983). Although D.
aciculare, the type species of Dissoconium, was originally assumed
to be hyperparasitic on powdery mildew (de Hoog et al. 1983),
Jackson et al. (2004) revealed that another species, D. dekkeri,
36
could act as a foliar pathogen of Eucalyptus. Dissoconium dekkeri
is, however, most commonly found in leaf spots in association
with other species of Teratosphaeria and Mycosphaerella.
Species of Dissoconium remain commensalists, and frequently
occur asexually on lesions associated with pathogenic species
of Capnodiales (Crous unpubl. data). They are ecologically
and morphologically quite distinct from other members of the
Capnodiales, and hence a separate family, the Dissoconiaceae,
is herewith introduced to accommodate them. Ramichloridium
forms brown, solitary conidiophores with a rachis and apical loci
similar to that observed on Dissoconium, and primary conidia that
are pale brown, 0–1-septate, with slightly thickened hila, but lacks
secondary conidia (Arzanlou et al. 2008b). Both Dissoconium and
Ramichloridium have in the past been reported as hyperparasitic
on powdery mildews on various hosts (Hijwegen & Buchenauer
1984), which suggests that they share a similar ecology.
Teratosphaeriaceae Crous & U. Braun, Stud. Mycol. 58: 8.
2007.
Type species: Teratosphaeria ibrillosa Syd. & P. Syd., Ann. Mycol.
10: 40. 1912.
Notes: Since the family was established by Crous et al. (2007a)
it has been shown to be too widely deined, incorporating many
diverse genera (Crous et al. 2009b, c), and even families such as
the Piedraiaceae (Fig. 1). The node as such is not well supported,
suggesting that as more taxa are added, further families remain to
be separated from the Teratosphaeriaceae. Presently it incorporates
diverse elements, and even lichens such as Cystocoleus ebeneus
and Anisomeridium consobrinum. The identity of the latter strain
(CBS 101364) needs to be conirmed, as its position in the tree
appears doubtful.
The genus Catenulostroma, which is associated with numerous
diverse substrates and habitats (Crous et al. 2007a), is typiied by C.
protearum, for which an epitype is designated in the present study.
Several strains isolated from rock surfaces (Guiedan et al. 2008,
Ruibal et al. 2008, 2009, this volume) cluster with Catenulostroma
(Fig. 1), and appear to represent undescribed species of the latter. Of
interest is the fact that the type species of Aulographina, A. pinorum
(CBS 302.71, 174.90), which has hysterothecia, clusters in a clade
with Catenulostroma microsporum, which has a Teratosphaeria-like
teleomorph with pseudothecia (Taylor & Crous 2000, Crous et al.
2004a, 2007a). Isolates of A. pinorum were found to produce a
Catenulostroma anamorph in culture. This raises two possibilities,
namely that either the incorrect fungus was originally isolated
from pine needles (namely Catenulostroma abietis), or that this is
a species complex, in which A. pinorum resides. If these strains
are indeed conirmed to represent A. pinorum, then it reveals
the genus Aulographina to be heterogeneous, as A. eucalypti,
which is a major leaf spot pathogen of Eucalyptus (Crous et al.
1989, Park et al. 2000, Carnegie & Keane 2003), clusters distant
from A. pinorum. The taxonomy of these taxa is currently being
addressed, and will be reported on elsewhere (Cheewangkoon et
al., in prep.). During the course of this study some new members of
the Teratosphaeriaceae were collected, which are described below:
PhylogenetiC lineages in the Capnodiales
fig. 7. Catenulostroma protearum. A. Colony on OA. B–G. Sporulating colony, with variable muriform to transversely septate conidia. Scale bars = 10 µm.
Catenulostroma protearum (Crous & M.E. Palm) Crous &
U. Braun, Stud. Mycol. 58: 17. 2007. Fig. 7.
The present species was one of a batch of novel taxa that Frank
collected and sent to us for treatment shortly before he had a
relapse. Frank’s friendship and mycological expertise will be sorely
missed.
Culture characteristics: On MEA spreading, erumpent, with folded
surface, and unevenly lobed, smooth margins; aerial mycelium
sparse; surface iron-grey to greenish black, reverse greenish black;
reaching 15 mm diam after 2 wk; similar on PDA and OA.
Devriesiae strelitziae similis, sed conidiis minoribus, (5–)7–10(–12) × (2–)2.5(–3)
µm.
Basionym: Trimmatostroma protearum Crous & M.E. Palm, Mycol.
Res. 103: 1303. 1999.
Host range and geographic distribution: Protea, Leucadendron and
Hakea spp., South Africa.
Specimens examined: South Africa, on leaves of Protea grandiceps, L. Schroeder,
15 Sept. 1986, holotype BPI 1107849; South Africa, Western Cape Province,
Stellenbosch, Assegaibos, on leaves of Leucadendron tinctum, F. Roets, 16 Apr.
2008, epitype designated here CBS H-20338, culture ex-epitype, CPC 15369,
15370 = CBS 125421; ditto, on leaves of Hakea sericea, CBS H-20339, single
ascospore culture CPC 15368.
Notes: Catenulostroma protearum was originally described from
dead leaves of Protea grandiceps collected in South Africa (Crous
& Palm 1999). Unfortunately the cultures died before they could be
deposited, and hence the phylogenetic position of Catenulostroma
remained uncertain. This proved to be problematic, as the genus
was later shown to be heterogeneous (Crous et al. 2007a).
The designation of the epitype in the present study clariies the
phylogenetic position of the genus, and reveals Catenulostroma
s. str. to represent species that occur in extreme environments,
on rocks, or on hard, leathery leaves such as Proteaceae and
Gymnospermae.
Devriesia hilliana Crous & U. Braun, sp. nov. MycoBank
MB514700. Fig. 8.
Etymology: Named in fond memory of Dr C.F. Hill. “Frank” collected
numerous fungi over the years, and sent them to the various
international colleagues he knew to be working on these groups.
www.studiesinmycology.org
Colonies sporulating on MEA. Mycelium consisting of branched,
septate, pale brown, smooth, 2–3 µm wide hyphae. Conidiophores
solitary, erect on creaping hyphae, unbranched, medium brown,
smooth, lexuous, thick-walled, 15–50 × 2–3 µm, 3–11-septate.
Conidiogenous cells terminal, medium brown, subcylindrical,
smooth, 5–20 × 2–3 µm; proliferating sympodially; hila lattened,
unthickened, somewhat darkened, 1–1.5 µm wide. Conidia
medium brown, smooth, subcylindrical to narrowly fusoid-ellipsoidal
or obclavate, apical conidium with obtuse apex, additional conidia
with truncate ends, somewhat darkened, 1–1.5 µm wide; conidia
straight to irregularly bent, mostly in unbranched chains, (5–)7–
10(–12) × (2–)2.5(–3) µm.
Culture characteristics: On MEA erumpent, spreading, with folded
surface, and smooth margins with sparse aerial mycelium; surface
mouse-grey, with thin, olivaceous-grey margin; reverse iron-grey,
reaching 8 mm diam; on PDA similar, up to 8 mm diam, centre
mouse-grey, margin and reverse iron-grey; on OA erumpent with
moderate mouse-grey aerial mycelium, and iron-grey margin.
Host range and geographic distribution: Macrozamia communis,
Auckland, New Zealand.
Specimen examined: New Zealand, Auckland, Auckland University Campus,
Princes Street, on Macrozamia communis, C.F. Hill, 20 Apr. 2008, CBS H-20340
holotype, culture ex-type CPC 15382 = CBS 123187.
37
Crous et al.
fig. 8. Devriesia hilliana. A. Sporulating colony on OA. B–D. Conidiophores giving rise to catenulate conidia. E–G. Fragmenting conidial segments from aerial hyphae. Scale
bars = 10 µm.
Devriesia lagerstroemiae Crous & M.J. Wingf., sp. nov.
MycoBank MB514701. Fig. 9.
patches of iron-grey; reverse iron-grey, reaching 10 mm diam; on
PDA similar, but on OA iron-grey, reaching 15 mm diam.
Etymology: Named after the host on which it occurs, Lagerstroemia.
Host range and geographic distribution: Lagerstroemia indica,
U.S.A., Louisiana.
Devriesiae strelitziae similis, sed conidiis latioribus, (5–)7–10(–12) × (2–)2.5(–3)
µm.
Colonies sporulating on OA. Mycelium consisting of smooth,
branched, septate, 2–3 µm wide hyphae. Conidiophores rarely
micronematous, predominantly macronematous, erect on creeping
hyphae, brown, cylindrical with swollen basal cell, thick-walled,
smooth, lexuous, 20–90 × 3–4 µm, 5–20-septate. Conidiogenous
cells terminal, cylindrical to clavate, polyblastic, pale to medium
brown, 5–10 × 2–3(–4) µm; scars somewhat thickened and
darkened, not refractive. Ramoconidia medium brown, smooth,
subcylindrical, 9–15 × 3–5 µm, (0–)1(–2)-septate, but with clavate
apex and several lattened loci that are somewhat darkened and
thickened, 1 µm diam. Conidia in branched chains of up to 10,
pale brown, smooth, narrowly ellipsoid, 0–1-septate, (5–)8–12(–15)
× 2–3(–4) µm; apical conidium with rounded apex, the rest with
lattened loci that are somewhat darkened and thickened, not
refractive, 0.5–1 µm diam.
Culture characteristics: On MEA erumpent, spreading, with sparse
aerial mycelium and irregular margin; surface olivaceous-grey, with
38
Specimen examined: u.S.A., Louisiana, Baton Rouge, Cod & Cook Centre,
N30°24’50.3” W91°10’6.6”, on Lagerstroemia indica, P.W. Crous & M.J. Wingield,
holotype CBS H-20341, culture ex-type CPC 14403 = CBS 125422.
Notes: Devriesia lagerstroemiae clusters close to D. hilliana.
As far as we know, neither species is heat-resistant, nor forms
chlamydospores, and hence the placement in Devriesia is more
due to phylogenetic similarity than their ecology.
Devriesia strelitziicola Arzanlou & Crous, sp. nov. MycoBank MB514702. Fig. 10.
Etymology: Named after its host plant, Strelitzia.
Devriesiae strelitziae similis, sed conidiis majoribus, (7–)25–45(–100) × (2–)2.5(–3)
µm.
Colonies sporulating on OA. Mycelium consisting of medium
brown, smooth, septate, branched, 2–3 µm wide hyphae;
chlamydospores not observed. Conidiophores dimorphic.
Microconidiophores reduced to conidiogenous cells on hyphae,
PhylogenetiC lineages in the Capnodiales
fig. 9. Devriesia lagerstroemiae. A. Leaves and lowers of Lagerstroemia indica. B. Leaf spots. C. Colony on OA. D–H. Conidiophores giving rise to branched conidial chains.
Scale bars = 10 µm.
erect, cylindrical, medium brown, smooth with truncate ends,
proliferating sympodially, 4–7 × 2–3 µm. Macroconidiophores erect,
cylindrical, straight to geniculate-sinuous, medium brown, smooth,
unbranched or branched above, 30–100 × 2.5–3 µm, 3–10-septate.
Conidiogenous cells terminal or lateral on branched conidiophores,
medium brown, smooth, cylindrical, proliferating sympodially, 7–15
× 2.5–3 µm; loci truncate, inconspicuous, 1–1.5 µm wide. Conidia
medium brown, smooth, guttulate, subcylindrical to narrowly
obclavate, apex obtuse to truncate, base truncate, occurring in
branched chains, widest at the basal septum, (7–)25–45(–100)
× (2–)2.5(–3) µm, (0–)3–6(–13)-septate; hila inconspicuous to
somewhat darkened and thickened, not refractive, 1–1.5 µm wide.
appears that a different generic name will have to be introduced to
accommodate those taxa occurring on plants. Further collections
are required, however, to clarify the generic boundaries of Devriesia
(Crous et al. 2007b).
Culture characteristics: On MEA erumpent, slow growing, with
moderate aerial mycelium and smooth margins; surface mousegrey, reverse iron-grey, reaching 8 mm diam after 2 wk; similar on
PDA and OA.
Colonies sporulating on MEA. Mycelium consisting of pale
brown, smooth, septate, branched, 3–4 µm wide hyphae that
become darker and thick-walled in the conidiogenous region.
Conidiogenous cells integrated, intercalary on hyphae, reduced
to short cylindrical loci, 2–2.5 µm wide, 1–4 µm tall; collarettes
inconspicuous to minute; proliferating 1–2 times percurrently at
apex. Conidia ellipsoid, aseptate, pale to medium brown, (4–)5–
7(–9) × (2.5–)3 µm, verruculose, apex obtuse, base subtruncate
with minute collarette; becoming swollen and elongate at maturity,
with 1–4 transverse and 1–2 oblique septa; (9–)10–13(–15) × (4–)
5–6(–9) µm; hila inconspicuous, up to 2 µm wide, frequently with
visible marginal frill; microcyclic conidiation commonly observed on
OA, MEA and PDA.
Host range and geographic distribution: Strelitzia sp., South Africa.
Specimen examined: South Africa, KwaZulu-Natal, Durban, Botanical Garden
near Reunion, on leaves of Strelitzia sp., 5 Feb. 2005, W. Gams & H. Glen, CBS
H-20342, holotype, culture ex-type X1045 = CBS 122480.
Notes: Devriesia strelitziicola is the second Devriesia species to
be described from this host (Arzanlou et al. 2008a). The genus
Devriesia was originally established to accommodate a group of
heat-resistant, Cladosporium-like fungi (Seifert et al. 2004), and it
www.studiesinmycology.org
Hortaea thailandica Crous & K.D. Hyde, sp. nov. MycoBank MB514703. Fig. 11.
Etymology: Named after the country where it was collected,
Thailand.
Hortaeae werneckii similis, sed conidiis brunneis, verruculosis, majoribus, (9–)10–
13(–15) × (4–)5–6(–7) µm.
39
Crous et al.
fig. 10. Devriesia strelitziicola. A. Strelitzia sp. with dead leaves. B. Colony on OA. C–G. Conidiophores giving rise to conidia. H–M. Conidia. Scale bars = 10 µm.
Culture characteristics: On MEA erumpent, spreading; surface
irregular, folded, greenish black, with sparse olivaceous-grey aerial
mycelium and smooth, lobed, margins; reverse greenish black;
reaching 12 mm diam after 2 wk; similar on OA and PDA.
Host range and geographic distribution: Syzygium siamense,
Thailand.
Specimen examined: Thailand, Khao Yai National Park, N14°14’42.6”
E101°22’15.7”, on leaves of Syzygium siamense, in lesions with a cercosporoid
fungus, 27 Mar. 2009, P.W. Crous & K.D. Hyde, holotype in BBH, isotype CBS
H-20343, culture ex-type CPC 16652, 16651 = CBS 125423, also in BCC.
Notes: Similar to Hortaea werneckii, which is also frequently
isolated from lesions in association with plant pathogenic
fungi, H. thailandica occurred in leaf spots in association with a
cercosporoid fungus. It is distinct from H. werneckii by forming
larger conidia that turn medium brown and verruculose with age.
40
Several other taxa are newly placed in the Teratosphaeriaceae
in the present study that require further evaluation. Xenomeris
juniperi, a bitunicate ascomycete on Jupinerus with pseudothecia
associated with a stroma, and pigmented, 1-septate ascospores,
clusters close to Teratosphaeria species occurring on Protea and
Eucalyptus, where the ascomata are also associated with stromatic
tissue (Taylor & Crous 2000, Crous et al. 2006c). Fresh collections
of this fungus would be required, however, to resolve its status. The
occurrence of Sporidesmium species in the Teratosphaeriaceae
should be interpreted with care, as the genus is polyphyletic, and
further studies are required to resolve its status (Shenoy et al.
2006, Crous et al. 2008a, Yang et al., in prep.).
PhylogenetiC lineages in the Capnodiales
fig. 11. Hortaea thailandica. A. Cercosporoid leaf spots on Syzygium siamense, in which H. thailandica occurred. B. Colonies on OA. C–E. Hyphae with conidiogenous loci
(arrows). F–H. Conidia. Scale bars = 10 µm.
Davidiellaceae C.L. Schoch, Spatafora, Crous & Shoemaker, Mycologia 98: 1048. 2006.
Type species: Davidiella tassiana (De Not.) Crous & U. Braun,
Mycol. Progr. 3: 8. 2003.
Notes: The Davidiellaceae was introduced for the genus Davidiella,
which has Cladosporium anamorphs. As shown in the present
analysis, however, allied genera such as Toxicocladosporium,
Verrucocladosporium, Rachicladosporium and Graphiopsis also
belong in this family. Of interest is the position of Melanodothis caricis
in Cladosporium s. str. This fungus, which infects lorets of Carex
and Kobresia, forms a stroma that gives rise to several immersed
ascomata with bitunicate, oblong asci that are aparaphysate, and
0–(2)-septate, hyaline, 9–14.5 × 2–4 µm ascospores. In culture,
a hyaline, Ramularia-like anamorph developed, with sympodial
proliferation, catenulate conidia, with thickened, darkened loci
(Arnold 1971). Although these characteristics are atypical of the
Davidiella/Cladosporium species in this clade, the position of
Melanodothis caricis in this family cannot simply be disregarded.
However, the ex-type culture of this fungus (CBS 860.72) proved
to be sterile.
A further unconirmed sequence (CBS 354.29, culture sterile,
but fast growing, grey-brown, Cladosporium-like), is that submitted
as Sphaerulina polyspora. The culture was accessioned in 1929,
deposited by A.E. Jenkins, and there is reason to believe that it
was derived from BPI 623724!, which is authentic for the species,
www.studiesinmycology.org
and collected by F.A. Wolf in May 1924. Wolf (1925) described this
fungus from twigs of Oxydendron arboretum with die-back disease
symptoms, collected in Raleigh, North Carolina. Sphaerulina
polyspora (623723 = Type!) has pseudothecia with aparaphysate,
bitunicate asci, and ascospores that are hyaline, 3–5-septate, 20–
24 × 6–7 µm. On the host it was linked to a Phoma-like anamorph,
which also grew similar in culture (yeast-like budding), and has
hyaline conidia which are ellipsoidal, 7–8 × 3.8–4 µm.
Colonies were reported as slow-growing, grey, appressed,
with germinating ascospores forming yeast-like budding cells, and
rarely having hyphae that extended from the margin of the colonies.
The link between Sphaerulina-like species, with Selenophoma
and Aureobasidium synanamorphs was recently illustrated
by Cheewangkoon et al. (2009). Although members of the
Dothideomycetes, these taxa do not cluster in the Davidiellaceae,
and hence it seems a fair assumption that CBS 354.29 is not
representative of Sphaerulina polyspora.
Rachicladosporium cboliae Crous, sp. nov. MycoBank
MB514704. Fig. 12.
Etymology: Named after the Consortium for the Barcode of Life,
CBOL, who organised a Fungal Barcoding Symposium, during
which this fungus was collected.
Rachicladosporio americano similis, sed conidiophoris dense fasciculatis et conidiis
minoribus.
41
Crous et al.
fig. 12. Rachicladosporium cboliae. A. Front Royal collection site in Virginia. B–E, G. Conidiophores with branched conidial chains. F. Hyphal coil. H–I. Chlamydospores in
chains. J. Conidia. Scale bars = 10 µm.
Colonies sporulating on OA. Mycelium consisting of branched,
septate hyphae, pale brown, smooth, 1.5–3 µm wide, frequently
constricted at septa, forming hyphal coils, but characteristically also
forming intercalary and terminal clusters of chlamydospores that
are brown, thick-walled, up to 6 µm diam. Conidiophores forming
laterally on creeping hyphae, erect, visible as densely branched
tufts on agar surface; conidiophores medium brown, smooth, thickwalled with bulbous base, lacking rhizoids, cylindrical, unbranched,
lexuous, up to 250 µm long, 4–6 µm wide, 10–20-septate.
Conidiogenous cells terminal, medium brown, smooth, polyblastic,
subcylindrical, 10–20 × 3–4 µm; loci terminal, thickened, darkend,
refractive, 1 µm diam. Ramoconidia 0(–1)-septate, subcylindrical,
medium brown, smooth, 7–12 × 3–4 µm. Conidia 0(–1)-septate, in
branched chains of up to 10, ellipsoid, pale brown, smooth, (6–)7–
8(–10) × (2–)2.5(–3) µm; hila thickened, darkened and refractive,
up to 1 µm diam.
Culture characteristics: On MEA spreading with sparse aerial
mycelium and smooth margins; surface folded, centre pale mousegrey to mouse-grey, margin iron-grey; reverse greenish black,
reaching 15–20 mm diam after 2 wk; on PDA spreading with
42
moderate aerial mycelium and smooth margins; surface olivaceousgrey, margin mouse-grey, reverse olivaceous-grey; reaching 30
mm diam; on OA spreading, folded with moderate aerial mycelium;
surface pale mouse-grey (centre) to olivaceous-grey at margin,
reaching 20 mm diam.
Host range and geographic distribution: Twig litter, Virginia, U.S.A.
Specimen examined: U.S.A., Virginia, Front Royal, N38°53’35” W78°10’50”, on twig
debris, 14 May 2007, P.W. Crous, holotype CBS H-20344, cultures ex-type CPC
14034 = CBS 125424, CPC 14035, 14036.
Notes: Rachicladosporium cboliae is a cryptic species close to R.
americanum, which was collected at the same site. They can be
distinguished on the litter in that R. cboliae has conidiophores with
densely branched tufts of conidia, in contrast to the more sparsely
branched conidiophores of R. americanum. Furthermore, R. cboliae
also forms prominent chains of chlamydospores in culture, which
lacks in R. americanum. Finally, R. cboliae has smaller ramoconidia
and conidia than those found in R. americanum (ramoconidia 13–23
× 3–4 µm; conidia 10–18 × 3–4 µm; Cheewangkoon et al. 2009).
PhylogenetiC lineages in the Capnodiales
DISCuSSIoN
The class Dothideomycetes incorporates fungal taxa exhibiting a
wide range of nutritional modes, and results in these fungi being
found in many diverse niches (Fig. 13). The two largest orders
Pleosporales (Zhang et al. 2009; this volume) and Capnodiales
encapsulate this diversity. Here we continue to expand sampling
within the Capnodiales in order to provide a well founded
phylogenetic scaffold for taxonomic classiication, informative
genomic sampling, ecological studies and evolutionary evaluations.
Capnodiales
The Capnodiales currently contain nine families (Lumbsch
& Huhndorf 2007, Kirk et al. 2008), a selection of which are
included in this study, namely Capnodiaceae, Davidiellaceae,
Mycosphaerellaceae, Piedraiaceae, and Teratosphaeriaceae.
Unfortunately, no cultures were available of the Antennulariellaceae
and Metacapnodiaceae, while Coccodiniaceae was again
shown to cluster outside the order, in Chaetothyriales (Crous
et al. 2007a). Families supported within Capnodiales (Fig. 1)
include Capnodiaceae, Davidiellaceae, Teratosphaeriaceae,
Dissoconiaceae, Schizothyriaceae and Mycosphaerellaceae. No
support was obtained for Piedraiaceae, which appeared to cluster
within Teratosphaeriaceae.
One of the main aims of the present study was to resolve
the status of the Capnodiales and Mycosphaerellales. Although
we were able to distinguish a clear, well resolved node for the
Mycosphaerellales (incl. Mycosphaerellaceae), this node was not
well supported, and elevating it to ordinal level would mean that
additional orders need to be introduced to accommodate several
families outside the Capnodiales s. str. This inding led us to
conclude that it is best to retain all families within a single, diverse
order, namely the Capnodiales.
evolution of nutritional modes and ecological
growth habits
The ancestral state of the present assemblage of taxa is likely
to be saprobic, as Phaeotheca (Sigler et al. 1981, de Hoog et al.
1997, Tsuneda et al. 2004), and Comminutispora (Ramaley 1996)
represent the earliest diverging lineages. This was similarly found
for a majority of lineages in the larger context of Ascomycota
(Schoch et al. 2009a, b). These taxa were not only all isolated from
dead materials or substrates, but they also share the same unique
mode of conidiogenesis, namely endoconidia, and a “black-yeast”
appearance in culture. Phaeotheca, which is strongly halophilic
(Zalar et al. 1999) is closely related to the lichen Racodium rupestre,
which forms an association with Trentepohlia algae, in which the
ilamentous algae is enclosed by melanised hyphae of the fungus.
This feature is also shared by another lichen, namely Cystocoleus
ebeneus (Teratosphaeriaceae) (Muggia et al. 2008). The
Capnodiaceae (sooty molds) that also cluster in a basal position in
the tree are epiphytes, growing on insect exudates (honey dew). The
Capnodiaceae are related to the Davidiellaceae, which represent
Cladosporium and allied genera. This family contains a wide range
of ecological adaptations, from primary plant pathogens, such as
Graphiopsis chlorocephala on Paeonia (Schubert et al. 2007a,
Braun et al. 2008), “Mycosphaerella” iridis on Iris (David 1997), to
taxa opportunistic on humans, Cladosporium bruhnei (Schubert et
al. 2007b), to halotolerant taxa, Cladosporium sphaerospermum
www.studiesinmycology.org
(Zalar et al. 2007, Dugan et al. 2008), to saprobes, C. herbarum, C.
cladosporioides (Schubert et al. 2007b).
The Teratosphaeriaceae contains several disjunct elements,
many of which may still eventually be removed from the family as
more taxa and additional sequence data are added, providing a
better resolution to some of these clades. In its widest sense, the
family contains lichens (Anisomeridium, Cystocoleus), saprobes
(Catenulostroma spp.), and halophilic, hyperhydrotic or lipophilic
species that have been reported from humans (Piedraia, Hortaea,
Penidiella, Stenella) (de Hoog et al. 2000, Bonifaz et al. 2008,
Plemenitaš et al. 2008), with the most derived clades tending to
contain plant pathogens (Readeriella, Teratosphaeria).
Dissoconiaceae is an early diverging lineage to the
Mycosphaerellaceae and Schizothyriaceae. Whereas most
members of Dissoconiaceae appear to be commensalists, there
is evidence that some species could be plant pathogenic (Jackson
et al. 2004), while the Schizothyriaceae contains epiphytes (Batzer
et al. 2007). The Mycosphaerellaceae contains species that
are biotrophic (Polythrincium; Simon et al. 2009), necrotrophic
plant pathogens (Brunneosphaerella, Cercospora, Dothistroma,
Pseudocercospora, Pseudocercosporella, Ramularia, and
Septoria), as well as some species that are saprobic (Passalora,
Pseudocercospora, Ramichloridium and Zasmidium; Arzanlou et
al. 2007), or endophytic (Pseudocercosporella endophytica; Crous
1998).
Within the Capnodiales, the positioning of saprobes such
as Phaeotheca and Comminutispora and the sooty moulds
(Capnodiaceae) may represent the more primitive state, from
where transitions occurred to more lichenised, saprobic, biotrophic
and nectrotrophic, plant pathogenic members of the order (Fig. 13).
This appears to mirror the other large and diverse order in the class,
the Pleosporales (Zhang et al. 2009; this volume). Lichenisation,
as well as the ability to be saprobic or plant pathogenic evolved
more than once, though the taxa in the later diverging clades of the
tree tend to be strictly nectrotrophic plant pathogens. This should
be interpreted with care, however, as Polythrincium is presently
the only biotrophic member included in this analysis, and other
biotrophic members of the Capnodiales may end up clustering
here, among the presently dominant nectrotropic plant pathogens.
One important and recent addition to Capnodiales diversity is
the rock-inhabiting fungi (Ruibal et al. 2008, 2009; this volume).
Although so far mainly isolated from sources in Antarctica and the
Mediterranean area, it is clear that they are a ubiquitous group of
fungi likely found throughout the globe. Their genetic diversity is
underscored by the fact that rock inhabiting fungi of convergent
morphology are also placed in other ascomycotan classes and
orders (Gueidan et al. 2008). The fact that many of these species
have reduced morphologies and are slow growers make their
taxonomy challenging, but their phylogenetic placement within
Teratosphaeriaceae and several other lineages within Capnodiales
makes their inclusion in subsequent phylogenetic assessments of
this order essential.
For this study, we designed novel primers to supplement
primers presently available in literature. Although primers are
usually designed for the genus or family of interest, they frequently
tend to have a wider application. Therefore, we attempted to design
our primers using a wide range of sequences from the GenBank
sequence database, in the hope that these primers will eventually
ind application outside of the Capnodiales as well. Although this
remains to be tested, we expect it to be the case. Our sequencing
of the complete SSU and LSU for the selected members of the
Capnodiales had a surprisingly large number of insertions present
43
Crous et al.
fig. 13. Members of Capnodiales exhibiting different ecological growth habits. A–C. Mycosphaerella marksii (plant pathogen). A. Leaf spot on Eucalyptus. B. Homothallic colony
on MEA. C. Asci. D. Conidiophore of Cladosporium sphaerospermum (saprobe). E–G. Ascomata and asci of Davidiella macrocarpa (saprobe). H–J. Dissoconium dekkeri (plant
pathogen, commensalist). H. Colony sporulating on MEA, with discharged conidia at the margin. I. Asci. J. Primary and secondary conidia attached to conidiophore. K–L.
Dissoconium proteae (commensalist). K. Sporulation on MEA with microsclerotia. L. Two conidial types attached to conidiophore (arrow). M–Q. Conidioxyphium gardeniorum
(sooty mold). M. Sporulation on MEA. N–P. Elongated, branched conidiomata with apical ostiolar hyphae. Q. Conidia. R–T. Leaf spot, ascus and verruculose ascospores of
Teratosphaeria ibrillosa (plant pathogen). U–X. Schizothyrium pomi (epiphyte). U. Thyrothecia occurring on a Rhus stem. V. Ascomatal initials forming on OA. W. Asci. X.
Conidiophore and conidia in vitro. Scale bars: E = 200 µm, M–O = 50 µm, all others = 10 µm.
44
PhylogenetiC lineages in the Capnodiales
for numerous strains. Although some of these insertions were
anticipated based on data already present in GenBank’s database,
the insertions in the LSU were not expected based on the sequences
used for primer design. However, this could be a result of the fewer
complete LSU sequences available in the database rather than a
deviation on the part of members of the Capnodiales. More complete
LSU sequences are needed from diverse orders to test whether
this is the case or not. Some of the taxa sequenced during this
study had insertions present at almost all of the possible insertion
positions, e.g. Mycosphaerella latebrosa, Septoria quercicola and
Teratosphaeria mexicana. These taxa are distributed throughout
the tree, and do not only cluster in a basal position, and therefore it
is dificult to predict why so many insertions were present. If these
insertions were all present in a basal position, it would have been
possible to argue that the higher number of insertions represents
the ancestral condition, and that these insertions are lost during
evolution. However, this proved not to be the case, and it could be
that these taxa accumulated these insertions.
Although the present study adds signiicantly to our knowledge
of the Capnodiales, the Capnodiaceae are still underrepresented,
and probably consist of numerous diverse lineages that can
be elevated to family level once our phylogenies become more
resolved. Regardless of this fact, the Mycosphaerellaceae clade
appears to be quite robust. It seems likely that further sampling of
the diverse Teratosphaeriaceae will necessitate further taxonomic
changes. The fact that the saprobic and plant pathogenic and
endophytic modes have evolved several times in different
families, suggest that many taxa can still easily adapt to changing
environments. A focus on adding more lichenicolous taxa, and taxa
occurring on non-plant substrates is crucial to provide further insight
into the ecological adaptations occurring in the Capnodiales.
ACKNowleDGemeNTS
Several colleagues have helped to collect material studied here, without which this
work would not have been possible. Drs E.H.C. Mckenzie and S.R. Pennycook
(Landcare New Zealand) are speciically thanked for recollecting Phaeophleospora
atkinsonii. We are grateful to BIOTEC and Dr Lily Eurwilaichitr (Director, Bioresources
unit, BIOTEC) for assisting with a collection trip in Thailand under the collaborative
BIOTEC-CBS memorandum of understanding. We thank Miss Marjan Vermaas for
preparing the photographic plates, and M. Starink-Willemse, and A. van Iperen, for
assistance with DNA sequencing and fungal cultures. Work performed for this paper
by the second author after 2008 was supported in part by the Intramural Research
Program of the NIH, National Library of Medicine. Before 2008 work was funded by
a grant from NSF (DEB-0717476). We are grateful to Drs Roland Kirschner, Alan J.
Phillips and Treena I. Burgess for their critical comments on this script. The views
expressed, however, are those of the authors.
refereNCeS
Andjic V, Barber PA, Carnegie AJ, Hardy GEStJ, Wingield MJ, Burgess TI (2007).
Phylogenetic reassessment supports accommodation of Phaeophleospora
and Colletogloeopsis from eucalypts in Kirramyces. Mycological Research 111:
1184–1198.
Aptroot A (2006). Mycosphaerella and its Anamorphs: 2. Conspectus of
Mycosphaerella. CBS Biodiversity Series 5, Utrecht, The Netherlands.
Arnold RH (1971). Melanodothis caricis, n. gen., n. sp. and “Hyalodothis? caricis”.
Canadian Journal of Botany 49: 2187–2196.
Arzanlou M, Crous PW (2006). Phaeosphaeriopsis musae. In: Fungal Planet – A
Global Initiative to promote the Study of Fungal Biodiversity (Crous PW, Seifert
KA, Samson RA, Hawksworth DL eds). CBS, Utrecht, Netherlands. Fungal
Planet No. 9.
Arzanlou M, Crous PW, Groenewald JZ (2008a). Devriesia strelitziae. In: Fungal
Planet – A Global Initiative to promote the Study of Fungal Biodiversity (Crous
PW, Seifert KA, Samson RA, Hawksworth DL eds). CBS, Utrecht, Netherlands.
Fungal Planet No. 22.
www.studiesinmycology.org
Arzanlou M, Groenewald JZ, Fullerton RA, Abeln ECA, Carlier J, et al. (2008b).
Multiple gene genealogies and phenotypic characters differentiate several
novel species of Mycosphaerella and related anamorphs on banana. Persoonia
20: 19–37.
Arzanlou M, Groenewald JZ, Gams W, Braun U, Shin H-D, Crous PW (2007).
Phylogenetic and morphotaxonomic revision of Ramichloridium and allied
genera. Studies in Mycology 58: 57–93.
Auld BA (1969). Incidence of damage caused by organisms which attack crofton
weed in the Richmond-Tweed region of New South Wales. Australian Journal
of Science 32: 163.
Barr ME (1987). Prodomus to class Loculoascomycetes. Published by the author,
Amherst, Massachusetts.
Batzer JC, Gleason ML, Harrington TC, Tiffany LH (2005). Expansion of the sooty
blotch and lyspeck complex on apples based on analysis of ribosomal DNA
gene sequences and morphology. Mycologia 97: 1268–1286.
Batzer JC, Mercedes Diaz Arias M, Harrington TC, Gleason ML, Groenewald JZ,
Crous PW (2007). Four species of Zygophiala (Schizothyriaceae, Capnodiales)
are associated with the sooty blotch and lyspeck complex on apple. Mycologia
100: 246–258.
Beilharz V, Pascoe I (2002). Two additional species of Verrucisporota, one with a
Mycosphaerella teleomorph, from Australia. Mycotaxon 82: 357–365.
Belding RD, Sutton TB, Blankenship SM, Young E (2000). Relationship between
apple fruit epicuticular wax and growth of Peltaster fructicola and Leptodontidium
elatius, two fungi that cause sooty blotch disease. Plant Disease 84: 767–772.
Bess HA, Haramoto FH (1958). Biological control of pamakani, Eupatorium
adenophorum, in Hawaii by a tephritid gall ly, Procecidochares utilis. 1. The life
history of the ly and its effectiveness in the control of the weed. In: Proceedings
of the Tenth International Congress of Entomology, Vol. 4 (Becker EC, ed.).
Canada, Ottawa, Mortimer: 543–548.
Bonifaz A, Badali H, Hoog GS de, Cruz M, Araiza J, et al. (2008). Tinea nigra by
Hortaea werneckii, a report of 22 cases from Mexico. Studies in Mycology 61:
77–82.
Braun U, Crous PW, Dugan F, Groenewald JZ, Hoog GS de (2003). Phylogeny and
taxonomy of Cladosporium-like hyphomycetes, including Davidiella gen. nov.,
the teleomorph of Cladosporium s.str. Mycological Progress 2: 3–18.
Braun U, Crous PW, Schubert K (2008). Taxonomic revision of the genus
Cladosporium s. lat. 8. Reintroduction of Graphiopsis (= Dichocladosporium)
with further reassessments of cladosporioid hyphomycetes. Mycotaxon 103:
207–216.
Carnegie AJ, Keane PJ (2003). Variation in severity of target spot, caused by
Aulographina eucalypti, in a eucalypt species and provenance trial in Victoria.
Australasian Plant Pathology 32: 393–402.
Cheewangkoon R, Crous PW, Hyde KD, Groenewald JZ, To-anan C (2008). Species
of Mycosphaerella and related anamorphs on Eucalyptus leaves from Thailand.
Persoonia 21: 77–91.
Cheewangkoon R, Groenewald JZ, Summerell BA, Hyde KD, To-anun C, Crous PW
(2009). Myrtaceae, a cache of fungal biodiversity. Persoonia 23: 55–85.
Cortinas M-N, Burgess T, Dell D, Xu D, Crous PW, Wingield BD, Wingield MJ
(2006b). First record of Colletogloeopsis zuluense comb. nov., causing a stem
canker of Eucalyptus in China. Mycological Research 110: 229–236.
Cortinas M-N, Crous PW, Wingield BD, Wingield MJ (2006a). Multi-gene
phylogenies and phenotypic characters distinguish two species within the
Colletogloeopsis zuluensis complex associated with Eucalyptus stem cankers.
Studies in Mycology 55: 133–146.
Crous PW (1998). Mycosphaerella spp. and their anamorphs associated with leaf
spot diseases of Eucalyptus. Mycologia Memoir 21: 1–170.
Crous PW (2009). Taxonomy and phylogeny of the genus Mycosphaerella and its
anamorphs. Fungal Diversity 38: 1–24.
Crous PW, Aptroot A, Kang JC, Braun U, Wingield MJ (2000). The genus
Mycosphaerella and its anamorphs. Studies in Mycology 45: 107–121.
Crous PW, Braun U (2003). Mycosphaerella and its anamorphs. 1. Names published
in Cercospora and Passalora. CBS Biodiversity Series 1: 1–571.
Crous PW, Braun U, Groenewald JZ (2007a). Mycosphaerella is polyphyletic.
Studies in Mycology 58: 1–32.
Crous PW, Braun U, Schubert K, Groenewald JZ (2007b). Delimiting Cladosporium
from morphologically similar genera. Studies in Mycology 58: 33–56.
Crous PW, Denman S, Taylor JE, Swart L, Palm ME (2004a). Cultivation and
diseases of Proteaceae: Leucadendron, Leucospermum and Protea. CBS
Biodiversity Series 2: 1–228.
Crous PW, Ferreira FA, Sutton BC (1997). A comparison of the fungal genera
Phaeophleospora and Kirramyces (coelomycetes). South African Journal of
Botany 63: 111–115.
Crous PW, Gams W, Stalpers JA, Robert V, Stegehuis G (2004b). MycoBank: an
online initiative to launch mycology into the 21st century. Studies in Mycology
50: 19–22.
Crous PW, Groenewald JZ, Mansilla JP, Hunter GC, Wingield MJ (2004c).
Phylogenetic reassessment of Mycosphaerella spp. and their anamorphs
occurring on Eucalyptus. Studies in Mycology 50: 195–214.
45
Crous et al.
Crous PW, Groenewald JZ, Summerell BA, Wingield BD, Wingield MJ (2009a).
Co-occurring species of Teratosphaeria on Eucalyptus. Persoonia 22: 38–48.
Crous PW, Groenewald JZ, Wingield MJ, Aptroot A (2003). The value of ascospore
septation in separating Mycosphaerella from Sphaerulina in the Dothideales: a
Saccardoan myth? Sydowia 55: 136–152.
Crous PW, Groenewald JZ, Wood AR (2008a). Sporidesmium knawiae. In: Fungal
Planet – A Global Initiative to promote the Study of Fungal Biodiversity (Crous
PW, Seifert KA, Samson RA, Hawksworth DL eds). CBS, Utrecht, Netherlands.
Fungal Planet No. 29.
Crous PW, Kang JC, Braun U (2001). A phylogenetic redeinition of anamorph
genera in Mycosphaerella based on ITS rDNA sequence and morphology.
Mycologia 93: 1081–1101.
Crous PW, Knox-Davies PS, Wingield MJ (1989). A summary of fungal leaf
pathogens of Eucalyptus and the diseases they cause in South Africa. South
African Forestry Journal 149: 9–16.
Crous PW, Liebenberg MM, Braun U, Groenewald JZ (2006a). Re-evaluating the
taxonomic status of Phaeoisariopsis griseola, the causal agent of angular leaf
spot of bean. Studies in Mycology 55: 163–173.
Crous PW, Palm ME (1999). Systematics of selected foliicolous fungi associated
with leaf spots of Proteaceae. Mycological Research 103: 1299–1304.
Crous PW, Schroers HJ, Groenewald JZ, Braun U, Schubert K (2006b).
Metulocladosporiella gen. nov. for the causal organism of Cladosporium
speckle disease of banana. Mycological Research 110: 264–275.
Crous PW, Summerell BA, Carnegie AJ, Mohammed C, Himaman W, Groenewald
JZ (2007c). Foliicolous Mycosphaerella spp. and their anamorphs on Corymbia
and Eucalyptus. Fungal Diversity 26: 143–185.
Crous PW, Summerell BA, Carnegie AJ, Wingield MJ, Groenewald JZ (2009b).
Novel species of Mycosphaerellaceae and Teratosphaeriaceae. Persoonia 23:
119–146.
Crous PW, Summerell BA, Carnegie AJ, Wingield MJ, Hunter GC, et al. (2009c).
Unravelling Mycosphaerella: do you believe in genera? Persoonia 23: 99–118.
Crous PW, Summerell BA, Mostert L, Groenewald JZ (2008b). Host speciicity and
speciation of Mycosphaerella and Teratosphaeria species associated with leaf
spots of Proteaceae. Persoonia 20: 59–86.
Crous PW, Verkley GJM, Groenewald JZ, Samson RA (eds) (2009d). Fungal
Biodiversity. CBS Laboratory Manual Series. Centraalbureau voor
Schimmelcultures, Utrecht, Netherlands.
Crous PW, Wingield MJ (1996). Species of Mycosphaerella and their anamorphs
associated with leaf blotch disease of Eucalyptus in South Africa. Mycologia
88: 441–458.
Crous PW, Wingield MJ, Mansilla JP, Alfenas AC, Groenewald JZ (2006c).
Phylogenetic reassessment of Mycosphaerella spp. and their anamorphs
occurring on Eucalyptus. II. Studies in Mycology 55: 99–131.
Crous PW, Wingield MJ, Park RF (1991). Mycosphaerella nubilosa a synonym of M.
molleriana. Mycological Research 95: 628–632.
Crous PW, Wood AR, Okada G, Groenewald JZ (2008c). Foliicolous microfungi
occurring on Encephalartos. Persoonia 21: 135–146.
David JC (1997). A contribution to the systematics of Cladosporium. Revision of the
fungi previously referred to Heterosporium. Mycological Papers 172: 1–157.
Dodd AP (1961). Biological control of Eupatorium adenophorum in Queensland.
Australian Journal of Science 23: 356–365.
Dugan FM, Braun U, Groenewald JZ, Crous PW (2008). Morphological plasticity in
Cladosporium sphaerospermum. Persoonia 21: 9–16.
Ellis MB (1976). More dematiaceous hyphomycetes. CAB, International Mycological
Institute, Surrey, Kew, U.K.
Gardes M, Bruns TD (1993). ITS primers with enhanced speciicity for basidiomycetes
- application to the identiication of mycorrhizae and rusts. Molecular Ecology
2: 113–118.
Gargas A, Taylor JW (1992). Polymerase chain reaction (PCR) primers for amplifying
and sequencing 18S rDNA from lichenized fungi. Mycologia 84: 589–592.
Gueidan C, Ruibal Villaseñor C, Hoog GS de, Gorbushina AA, Untereiner WA,
Lutzoni F (2008). A rock-inhabiting ancestor for mutualistic and pathogen-rich
fungal lineages. Studies in Mycology 61: 111–119.
Hawksworth DL, Kirk PM, Sutton BC, Pegler DN (1995). Ainsworth and Bisby’s
Dictionary of the Fungi, 8th edn. CAB International, Wallingford, U.K.
Hernández-Gutiérrez A, Dianese JC (2009). New cercosporoid fungi from the
Brazilian Cerrado 2. Species on hosts of the subfamilies Caesalpinioideae,
Faboideae and Mimosoideae (Leguminosae s. lat.). Mycotaxon 107: 1–24.
Hijwegen T, Buchenauer H (1984). Isolation and identiication of hyperparasitic
fungi associated with Erysiphaceae. Netherlands Journal of Plant Pathology
90: 79–84.
Hoog GS de, Beguin H, Batenburg-van de Vegte WH (1997). Phaeotheca
triangularis, a new meristematic black yeast from a humidiier. Antonie van
Leeuwenhoek 71: 289–295.
Hoog GS de, Gerrits van den Ende AHG (1998). Molecular diagnostics of clinical
strains of ilamentous Basidiomycetes. Mycoses 41: 183–189.
Hoog GS de, Guarro J, Gené J, Figueras MJ (2000). Atlas of Clinical Fungi. 2nd Edn.
Centraalbureau voor Schimmelcultures, Utrecht, Netherlands, and Universitat
Rovira I Virgili, Reus, Spain.
46
Hoog GS de, Oorschot CAN van, Hijwegen T (1983). Taxonomy of the Dactylaria
complex. II. Proceedings van de Koninklijke Nederlandse Akademie van
Wetenschappen, Series C, 86(2): 197–206.
Hughes SJ. 1976. Sooty molds. Mycologia 68: 451–691.
Hunter GC, Wingield BD, Crous PW, Wingield MJ (2006). A multi-gene phylogeny
for species of Mycosphaerella occurring on Eucalyptus leaves. Studies in
Mycology 55: 147–161.
Jackson SL, Maxwell A, Neumeister-Kemp HG, Dell B, Hardy GEStJ (2004).
Infection, hyperparasitism and conidiogenesis of Mycosphaerella lateralis on
Eucalyptus globulus in Western Australia. Australasian Plant Pathology 33:
49–53.
Kirk PM, Cannon PF, David JC, Stalpers JA (2001). Ainsworth and Bisby’s Dictionary
of the Fungi, 9th edn. CAB International, Wallingford, U.K.
Kirk PM, Cannon PF, Minter DW, Stalpers JA (2008). Ainsworth and Bisby’s
Dictionary of the Fungi, 10th edn. CAB International, Wallingford, U.K.
Kluge RL (1991). Biological control of crofton weed, Ageratina adenophora
(Asteraceae), in South Africa. Agriculture, Ecosystems & Environment 37:
187–191.
Knox-Davies PS, Wyk PS van, Marasas WFO (1987). Diseases of Protea,
Leucospermum and Leucadendron recorded in South Africa. Phytophylactica
19: 327–337.
Kretzer A, Li Y, Szaro T, Bruns TD (1996). Internal transcribed spacer sequences
from 38 recognized species of Suillus sensu lato: phylogenetic and taxonomic
implications. Mycologia 88: 776–785.
Kruys A, Eriksson OE, Wedin M (2006). Phylogenetic relationships of coprophilous
Pleosporales (Dothideomycetes, Ascomycota), and the classiication of some
bitunicate taxa of unknown position. Mycological Research 110: 527–536.
Lumbsch HT, Huhndorf S (2007). Outline of Ascomycota. Myconet 13: 1–99.
Marincowitz S, Crous PW, Groenewald JZ, Wingield MJ (2008). Microfungi
occurring on Proteaceae in the fynbos. CBS Biodiversity Series 7: 1–166.
Masuka J, Cole DL, Mguni C (1998). List of Plant Diseases in Zimbabwe.
Department of Research Specialist Services, Ministry of Lands and Agriculture,
Harare, Zimbabwe.
Moncalvo J-M, Rehner SA, Vilgalys R (1993). Systematics of Lyophyllum section
Difformia based on evidence from culture studies and ribosomal DNA
sequences. Mycologia 85: 788–794.
Morris MJ (1989). Host speciicity studies of leaf spot fungus, Phaeoramularia sp. for
the biological control of crofton weed (Ageratina adenophorum) in South Africa.
Phytophylactica 21: 281–283.
Moura MF, Rodrigues PF (2001). Fungal diseases on Proteas identiied in Maderia
Island. Acta Horticulturae 545: 265–268.
Muggia L, Hafellner J, Wirtz N, Hawksworth DL, Grube M (2008). The sterile
microilamentous lichens Cystocoleus ebeneus and Racodium rupestre are
relatives of clinically important dothidealean fungi. Mycological Research 112:
50–56.
Muniappan R, Raman A, Reddy GVP (2009) Ageratina adenophora (Sprengel)
King and Robinson (Asteraceae). In: Biological control of tropical weeds using
arthropods. (Muniappan R, Reddy GVP, Raman A, eds). Cambridge University
Press, Cambridge: 1–16.
Park RF, Keane PJ, Wingield MJ, Crous PW (2000). Fungal diseases of eucalypt
foliage. In: Diseases and pathogens of eucalypts. (Keane PJ, Kile GA, Podger
FD, Brown BN, eds). CSIRO publishing, Australia: 153–239.
Parsons WT, Cuthbertson EG (1992). Noxious Weeds of Australia. Melbourne:
Inkata Press.
Pennycook SR, McKenzie EHC (2002). Scoleciasis atkinsonii, an earlier name for
Phaeophleospora hebes; and a note on G.H. Cunningham’s epithets hebe and
pseudopanax. Mycotaxon 82: 145–146.
Plemenitaš A, Vaupotič T, Lenassi M, Kogej T, Gunde-Cimerman N (2008).
Adaptation of extremely halotolerant black yeast Hortaea werneckii to
increased osmolarity: a molecular perspective at a glance. Studies in Mycology
61: 67–75.
Ramaley AW (1996). Comminutispora gen. nov. and its Hyphospora gen. nov.
anamorph. Mycologia 88: 132–136.
Rayner RW (1970). A mycological colour chart. CMI and British Mycological Society.
Kew, Surrey, England.
Rehner SA, Samuels GJ (1994). Taxonomy and phylogeny of Gliocladium analysed
from nuclear large subunit ribosomal DNA sequences. Mycological Research
98: 625–634.
Ruibal C, Platas G, Bills GF (2008). High diversity and morphological convergence
among melanised fungi from rock formations in the Central Mountain System
of Spain. Persoonia 21: 93–110.
Ruibal C, Gueidan C, Selbmann L, Gorbushina A, Crous PW, et al. (2009).
Phylogeny of rock-inhabiting fungi related to Dothideomycetes. Studies in
Mycology 64: 123–133.
Schoch CL, Shoemaker RA, Seifert KA, Hambleton S, Spatafora JW, Crous PW
(2006). A multigene phylogeny of the Dothideomycetes using four nuclear loci.
Mycologia 98: 1041–1052.
PhylogenetiC lineages in the Capnodiales
Schoch CL, Crous PW, Groenewald JZ, Boehm EWA, Burgess TI, et al. (2009a). A
class-wide phylogenetic assessment of Dothideomycetes. Studies in Mycology
64: 1–15.
Schoch CL, Sung GH, Lopez-Giraldez F, Townsend JP, Miadlikowska J, et al.
(2009b). The Ascomycota Tree of Life: A Phylum-wide Phylogeny Clariies
the Origin and Evolution of Fundamental Reproductive and Ecological Traits.
Systematic Biology 58: 224–239.
Schubert K, Braun U, Groenewald JZ, Crous PW (2007a). Cladosporium leaf-blotch
and stem rot of Paeonia spp. caused by Dichocladosporium chlorocephalum
gen. nov. Studies in Mycology 58: 95–104.
Schubert K, Groenewald JZ, Braun U, Dijksterhuis J, Starink M, et al. (2007b).
Biodiversity in the Cladosporium herbarum complex (Davidiellaceae,
Capnodiales), with standardisation of methods for Cladosporium taxonomy
and diagnostics. Studies in Mycology 58: 105–156.
Seifert KA, Nickerson NL, Corlett M, Jackson ED, Louis-Seize G, Davies RJ
(2004). Devriesia, a new hyphomycete genus to accommodate heat-resistant,
cladosporium-like fungi. Canadian Journal of Botany 82: 914–926.
Sharma KC, Chhetri GKK (1977). Reports on studies on the biological control of
Eupatorium adenophorum. Nepalese Journal of Agriculture 12: 135–157.
Shenoy BD, Jeewon R, Wu WP, Bhat DJ, Hyde KD (2006). Ribosomal and RPB2
DNA sequence analyses suggest that Sporidesmium and morphologically
similar genera are polyphyletic. Mycological Research 110: 916–928.
Sigler L, Tsuneda A, Carmichael JW (1981). Phaeotheca and Phaeosclera, two new
genera of dematiaceous hyphomycetes and a redescription of Sarcinomyces
Lindner. Mycotaxon 12: 449–467.
Simon UK, Groenewald JZ, Crous PW (2009). Cymadothea trifolii, an obligate
biotrophic leaf parasite of Trifolium, belongs to Mycosphaerellaceae as shown
by nuclear ribosomal DNA analyses. Persoonia 22: 49–55.
Singh SK, Chaudhary RK, Morgan-Jones G (1995). Notes on Hyphomycetes: LXVII.
Three new species of Phaeoramularia from Nepal. Mycotaxon 54: 57–66.
Spatafora JW, Mitchell TG, Vilgalys R (1995). Analysis of genes encoding for smallsubunit rRNA sequences in studying phylogenetics of dematiaceous fungal
pathogens. Journal of Clinical Microbiology 33: 1322–1326.
Stamatakis A (2006). RAxML-VI-HPC: maximum likelihood-based phylogenetic
analyses with thousands of taxa and mixed models. Bioinformatics 22: 2688–
2690.
Stamatakis A, Hoover P, Rougemont J (2008). A Rapid Bootstrap Algorithm for the
RAxML Web Servers. Systematic Biology 57: 758–771.
Taylor JE, Cannon PF, David JC, Crous PW (2001a). Two new Phaeophleospora
species associated with leaf spots of Proteaceae. South African Journal of
Botany 67: 39–43.
Taylor JE, Crous PW (1998). Leptosphaeria protearum. IMI Descriptions of Fungi
and Bacteria No. 1343.
Taylor JE, Crous PW (2000). Fungi occurring on Proteaceae. New anamorphs for
Teratosphaeria, Mycosphaerella and Lembosia, and other fungi associated
with leaf spots and cankers of Proteaceous hosts. Mycological Research 104:
618–636.
Taylor JE, Crous PW, Palm ME (2001b). Foliar and stem fungal pathogens of
Proteaceae in Hawaii. Mycotaxon 78: 449–490.
Thomma BPHJ, van Esse PH, Crous PW, Wit PJGM de (2005). Cladosporium
fulvum (syn. Passalora fulva), a highly specialized plant pathogen as a model
for functional studies on plant pathogenic Mycosphaerellaceae. Molecular
Plant Pathology 6: 379–393.
Tsuneda A, Tsuneda I, Currah RS (2004). Endoconidiogenesis in Endoconidioma
populi and Phaeotheca issurella. Mycologia 96: 1134–1140.
Vilgalys R, Hester M (1990). Rapid genetic identiication and mapping of
enzymatically ampliied ribosomal DNA from several Cryptococcus species.
Journal of Bacteriology 172: 4238–4246.
Wagner WL, Herbst DR, Sohmer SH (1999). Manual of the Flowering Plants of
Hawaii. Revised edition. Honolulu, HI: University of Hawaii Press.
Walker J, Sutton BC, Pascoe IG (1992). Phaeoseptoria eucalypti and similar
fungi on Eucalyptus with description of Kirramyces gen. nov. (coelomycetes).
Mycological Research 96: 911–924.
Wang F, Summerell BA, Marshall D, Auld BA (1997). Biology and pathology of a
species of Phaeoramularia causing a leaf spot of crofton weed. Australasian
Plant Pathology 26: 165–172.
White TJ, Bruns T, Lee S, Taylor J (1990). Ampliication and direct sequencing of
fungal ribosomal RNA genes for phylogenetics. In: PCR Protocols: a guide to
methods and applications (Innis MA, Gelfand DH, Sninsky JJ, White TJ, eds).
Academic Press, San Diego, California: 315–322.
Wolf FA (1925). Some undescribed fungi on sourwood, Oxydendron arboretum (L.)
DC. Journal of the Elisha Mitchell Scientiic Society 41: 94–99.
Wu W, Sutton BC, Gange AC (1996). Revision of Septoria species on Hebe and
Veronica and description of Kirramyces hebes sp. nov. Mycological Research
100: 1207–1217.
Yen JM, Lim G (1980). Cercospora and allied genera of Singapore and the Malay
Peninsula. Gardens’ Bulletin Singapore 33: 151–263.
www.studiesinmycology.org
Zalar P, Hoog GS de, Gunde-Cimerman N (1999). Ecology of halotolerant
dothideaceous black yeast. Studies in Mycology 43: 38–48.
Zalar P, Hoog GS de, Schroers H-J, Crous PW, Groenewald JZ, Gunde-Cimerman
N (2007). Phylogeny and ecology of the ubiquitous saprobe Cladosporium
sphaerospermum, with descriptions of seven new species from hypersaline
environments. Studies in Mycology 58: 157–183.
Zhang Y, Schoch CL, Fournier J, Crous PW, Gruyter J de, et al. (2009). Multi-locus
phylogeny of the Pleosporales: a taxonomic, ecological and evolutionary reevaluation. Studies in Mycology 64: 85–102.
Zhu L, Sun OJ, Sang W, Li Z, Ma K (2007). Predicting the spatial distribution of
an invasive plant species (Eupatorium adenophorum) in China. Landscape
Ecology 22: 1143–1154.
47
Crous et al.
SupplemeNTAry INformATIoN
Table 1. Details of the isolates for which novel sequences were generated. Samples without an 18S rDNA accession number were only
used in the 28S rDNA analysis; sequences of CBS 723.79 and CBS 123.26 were used in both analyses. The accession number for 5.8S
nrDNA also includes the lanking spacer regions.
Species
Accession number1 Host
Country
Collector
GenBank Accession numbers
18S nrDNA, 5.8S nrDNA, 28S
nrDNA
CBS 302.71; ETH 7129;
UAMH 4037
Aulographina pinorum
Pinus maritima
France
E. Müller
—, GU214622, GU214393
Batcheloromyces leucadendri CBS 110892; CPC 1837
Leucadendron sp.
South Africa
L. Swart
GU214515, AY260100, EU019246
Batcheloromyces proteae
CBS 110696; CPC 1518
Protea cynaroides
South Africa
L. Viljoen
AY251102, AY260099, EU019247
Brunneosphaerella
protearum
CPC 13905
Protea sp.
South Africa
P.W. Crous
—, GU214623, GU214394
CPC 13914
Protea sp.
South Africa
P.W. Crous
—, GU214624, GU214395
CPC 15231
Protea nitida
South Africa
L. Mostert
—, GU214625, GU214396
CPC 16338
Protea sp.
South Africa
P.W. Crous
—, GU214626, GU214397
CBS 214.90; CBS 176.88;
IAM 13014; JCM 6932;
TNS F-198506
Capnobotrys neessii
Japan
J. Sugiyama
AY220612, AY220612, GU214398
CBS 215.90; IAM 13015
Capnobotrys neessii
Japan
J. Sugiyama
AY220613, AY220613, GU214399
Capnobotryella renispora
Capnodium coffeae
CBS 147.52
Coffea robusta
Zaire
—
DQ247808, AJ244239, GU214400
Catenulostroma
chromoblastomycosum
CBS 597.97
Man,
chromoblastomycosis
Zaire
V. de Brouwere
GU214516, AJ244260, EU019251
Catenulostroma elginense
CBS 111030; CPC 1958
Protea grandiceps
South Africa
J.E. Taylor
GU214517, AY260093, EU019252
Catenulostroma germanicum
CBS 539.88
Stone
Germany
—
GU214518, EU019253, EU019253
Catenulostroma microsporum
CBS 110890; CPC 1832
Protea cynaroides
South Africa
L. Swart
GU214520, AY260097, EU019255
Catenulostroma protearum
CBS 125421; CPC 15370
Leucadendron tinctum
South Africa
F. Roets
—, GU214627, GU214401
CPC 15368
Hakea sericea
South Africa
F. Roets
—, GU214628, GU214402
CPC 15369
Leucadendron tinctum
South Africa
F. Roets
—, GU214629, GU214403
Cercospora apii
CBS 118712
—
Fiji
P. Tyler
GU214653, GU214653, GU214653
Cercospora beticola
CBS 116456; CPC 11557
Beta vulgaris
Italy
V. Rossi
AY840527, AY840527, GU214404
Cercospora capsici
CPC 12307
Capsicum annuum
South Korea
H.D. Shin
GU214654, GU214654, GU214654
Cercospora janseana
CBS 145.37; CPC 4303;
IMI 303642
Oryza sativa
U.S.A.
E.C. Tullis
AY251103, AY260064, GU214405
Cercospora sojina
CPC 12322
Glycine soja
South Korea
H.D. Shin
GU214655, GU214655, GU214655
Cercospora zebrinae
CBS 112893; CPC 3955
Trifolium protense
Canada
K. Seifert
AY251104, AY260078, GU214406
CBS 118789; WAC 5106
Trifolium subterraneum
Australia
M.J. Barbetti
GU214656, GU214656, GU214656
CBS 118790; IMI 262766;
WAC 7973
Trifolium subterraneum
Australia
M.J. Barbetti
GU214657, GU214657, GU214657
Cercosporella virgaureae
CBS 113304
Erigeron annueus
South Korea
H.D. Shin
GU214658, GU214658, GU214658
Cladosporium bruhnei
CBS 115683; ATCC
66670; CPC 5101
CCA-treated Douglas-ir
pole
U.S.A.
—
AY251096, AY251078, GU214408
CBS 188.54; ATCC
11290; IMI 049638; CPC
3686
—
—
G.A. de Vries
AY251098, AY251077, EU019263
Cladosporium
cladosporioides
CBS 109.21; ATCC
11277; ATCC 200940;
CPC 3682; IFO 6368; IMI
049625
Hedera helix
U.K.
G.A. de Vries
AY251093, AY251073, EU019262
CBS 401.80; CPC 3683
Triticum aestivum
Netherlands
N.J. Fokkema
AY251091, AY251074, GU214409
Cladosporium herbarum
CBS 723.79
Allium porrum
New Zealand
A.C. Jamieson
EU167558, EU167558, GU214410
Cladosporium sp.
CPC 15513
Rubus fruticosus
Italy
P.W. Crous
—, GU214630, GU214411
CPC 15516
Pyrus communis
Ukraine
A. Akulov
—, GU214631, GU214412
Cladosporium uredinicola
ATCC 46649; CPC 5390
Quercus nigra
U.S.A.
G. Morgan-Jones
AY251097, AY251071, EU019264
Davidiella rosigena
CBS 330.51
Leaf spot in Rosa sp.
Netherlands
—
—, GU214632, GU214413
www.studiesinmycology.org
47-S1
PhylogenetiC lineages in the Capnodiales
Table 1. (Continued).
Species
Accession number1 Host
Country
Collector
GenBank Accession numbers
18S nrDNA, 5.8S nrDNA, 28S
nrDNA
Devriesia hilliana
CBS 123187; CPC 15382
Macrozamia communis
New Zealand
C.F. Hill
—, GU214633, GU214414
Devriesia lagerstroemiae
CBS 125422; CPC 14403
Lagerstroemia indica
U.S.A.
P.W. Crous &
M.J. Wingield
—, GU214634, GU214415
Devriesia staurophora
CBS 375.81; ATCC
200934; CPC 3687
Páramo soil
Colombia
H. Valencia
EF137359, AF393723, GU214416
Devriesia strelitziicola
CBS 122480; X1045
Strelitzia sp.
South Africa
W. Gams &
H. Glen
—, GU214635, GU214417
Dissoconium aciculare
CBS 201.89
Brassica sp.
Netherlands
T. Hijwegen
GU214522, AY725519, GU214418
CBS 204.89
Astragalus sp.
Germany
T. Hijwegen
GU214523, AY725520, GU214419
CBS 342.82; CPC 1534
Erysiphe, on Medicago
lupulina
Germany
T. Hijwegen
GU214524, AF173308, EU019266
CBS 110747; CPC 831
Eucalyptus nitens
South Africa
P.W. Crous
GU214525, AY725535, GU214420
CBS 114238; CPC 10440
Eucalyptus globulus
Spain
J.P.M. Vazquez
GU214526, AY725541, EU019267
Dissoconium commune
Dissoconium dekkeri
Dothistroma pini
CBS 114239; CPC 10492
Eucalyptus globulus
New Zealand
W. Gams
GU214527, AY725542, GU214421
CBS 110748; CMW
14906; CPC 825
Eucalyptus grandis
South Africa
G. Kemp
GU214528, AF309625, GU214422
CBS 111169; CMW 5164;
CPC 1232
Eucalyptus globulus
Zambia
—
GU214529, AY725550, GU214423
CBS 111272; CPC 1188
Eucalyptus nitens
South Africa
M.J. Wingield
GU214530, AY725551, GU214424
CBS 111282; CPC 1233
Eucalyptus globulus
Zambia
—
GU214531, AF173305, GU214425
CBS 567.89; CPC 1535
Juniperus chinensis
Netherlands
T. Hijwegen
AY251101, AF173309, EU019268
CBS 116487; CMW 10951
Pinus nigra
U.S.A.
G. Adams
GU214532, AY808302, GU214426
Dothistroma septosporum
CBS 112498; CPC 3779
Pinus radiata
Ecuador
—
GU214533, AY293062, GU214427
Graphiopsis chlorocephala
CBS 121523; CPC 11969
Paeonia oficinalis
Germany
K. Schubert
GU214534, EU009458, EU009458
Hortaea acidophila
CBS 113389
Lignite, pH 1
Germany
U. Hölker
—, GU214636, GU214428
Hortaea thailandica
CBS 125423; CPC 16651
Syzygium siamense
Thailand
P.W. Crous &
K.D. Hyde
—, GU214637, GU214429
Lecanosticta acicola
CBS 871.95; MPFN 314
Pinus radiata
France
M. Morelet
GU214663, GU214663, GU214663
Leptoxyphium fumago
CBS 123.26; ATCC
11925; IMI 089363; LSHB
X13
Hibiscus tiliaceus
Indonesia
—
GU214535, —, GU214430
Melanodothis caricis
CBS 860.72; ATCC
24309; DAOM 116433
Carex sitchensis
Canada
—
—, GU214638, GU214431
Miuraea persicae
CPC 10069
Prunus persica
South Korea
H.D. Shin
GU214660, GU214660, GU214660
Mycosphaerella acaciigena
CBS 112515; CPC 3837
Acacia mangium
Venezuela
M.J. Wingield
AY251116, AY752143, GU214432
CBS 112516; CPC 3838
Acacia mangium
Venezuela
M.J. Wingield
GU214661, GU214661, GU214661
Mycosphaerella africana
CBS 116154; CMW 4945;
CPC 794
Eucalyptus viminalis
South Africa
P.W. Crous
GU214536, AF173314, GU214433
Mycosphaerella bixae
CBS 111804; CPC 2554
Bixa orellana
Brazil
P.W. Crous &
R.L. Benchimol
GU214557, AF362056, GU214455
Mycosphaerella ellipsoidea
CBS 110843; CPC 850
Eucalyptus cladocalyx
South Africa
P.W. Crous
GU214537, AY725545, GU214434
Mycosphaerella endophytica
CBS 114662; CPC 1193
Eucalyptus sp.
South Africa
P.W. Crous
GU214538, DQ302953, GU214435
Mycosphaerella graminicola
CBS 100335; IPO
69001.61
Triticum aestivum
—
G.H.J. Kema
GU214539, EU019297, EU019297
CBS 110744; CPC 658
Triticum sp.
South Africa
P.W. Crous
AY251117, AF362068, EU019298
CBS 115943; IPO323
Triticum aestivum
Netherlands
R. Daamen
GU214540, AF181692, GU214436
Mycosphaerella handelii
CBS 113302
Rhododendron sp.
Netherlands
P.W. Crous &
U. Braun
EU167581, EU167581, GU214437
Mycosphaerella heimii
CBS 110682; CMW 4942;
CPC 760
Eucalyptus sp.
Madagascar
P.W. Crous
GU214541, AF309606, GU214438
Mycosphaerella heimioides
CBS 111190; CMW 3046;
CPC 1312
Eucalyptus sp.
Indonesia
M.J. Wingield
GU214542, AF309609, GU214439
Mycosphaerella holualoana
CBS 110699; CPC 2155
Leucospermum sp.
U.S.A.: Hawaii
P.W. Crous
GU214543, AY260084, GU214440
47-S2
Crous et al.
Table 1. (Continued).
Species
Accession number1 Host
Country
Collector
GenBank Accession numbers
18S nrDNA, 5.8S nrDNA, 28S
nrDNA
Mycosphaerella
irregulariramosa
CBS 111211; CPC 1362
Eucalyptus saligna
South Africa
M.J. Wingield
GU214544, AF309608, GU214441
Mycosphaerella keniensis
CBS 111001; CMW 5147;
CPC 1084
Eucalyptus grandis
Kenya
M.J. Wingield
GU214545, AF173300, GU214442
Mycosphaerella latebrosa
CBS 652.85
Acer pseudoplatanus
Netherlands
H.A. van der Aa
AY251114, AF362067, GU214443
CBS 687.94
Acer pseudoplatanus
Netherlands
G. Verkley
GU214546, AY152553, GU214444
Mycosphaerella lupini
CPC 1661
Lupinus sp.
U.S.A.
W. Kaiser
GU214547, AF362050, FJ839661
Mycosphaerella marasasii
CBS 110790; CPC 348
Syzygium cordatum
South Africa
M.J. Wingield
GU214548, AF309591, GU214445
Mycosphaerella marksii
CBS 110942; CPC 982
Eucalyptus botryoides
Australia
A.J. Carnegie
GU214549, AF309589, GU214446
CPC 11222
Eucalyptus grandis
Bolivia
M.J. Wingield
GU214550, DQ302983, GU214447
Mycosphaerella parkii
CBS 387.92; CMW
14775; CPC 353
Eucalyptus grandis
Brazil
M.J. Wingield
GU214551, AF309590, GU214448
Mycosphaerella sp.
CBS 111166; CPC 1224
Eucalyptus cladocalyx
South Africa
A.R. Wood
GU214552, AF173302, GU214449
CBS 111167; CPC 1225
Eucalyptus cladocalyx
South Africa
A.R. Wood
GU214553, AF309593, GU214450
Mycosphaerella sphaerulinae
CBS 112621; CPC 4314
Eucalyptus sp.
Chile
—
GU214554, AY293066, GU214451
Mycosphaerella stromatosa
CBS 101953; CPC 1731
Protea sp.
South Africa
S. Denman
AY251115, EU167598, EU167598
Mycosphaerella tasmaniensis
CBS 111687; CMW
14780; CPC 1555
Eucalyptus nitens
Australia
—
GU214555, AF310107, GU214452
Passalora ageratinae
CBS 125419; CPC 15365
Ageratina adenophora
South Africa
A.R. Wood
—, GU214639, GU214453
Passalora bellynckii
CBS 150.49; CPC 3635
Cynanchum
vincetoxicum
Switzerland
S. Blumer
GU214556, AF222831, GU214454
Passalora brachycarpa
CBS 115124
—
—
C.F. Hill
GU214664, GU214664, GU214664
Passalora armatae
CBS 125420; CPC 15419
Dalbergia armata
South Africa
A.R. Wood
—, GU214640, GU214456
Passalora dioscoreae
CPC 10855
Dioscorea tokora
South Korea
H.D. Shin
GU214665, GU214665, GU214665
Passalora dodonaea
CPC 1223
Dodonaea sp.
—
P.W. Crous
AY251108, GU214641, GU214457
Passalora eucalypti
CBS 111318; CPC 1457
Eucalyptus saligna
Brazil: Suzano
P.W. Crous
GU214558, AF309617, GU214458
Passalora fulva
CBS 119.46; CPC 3688
Lycopersicon esculentum
Netherlands
—
AY251109, AY251069, DQ008163
Passalora graminis
CBS 113303
Alopecurus aequalis var.
amurensis
South Korea
H.D. Shin
GU214666, GU214666, GU214666
Passalora perplexa
CBS 116364; CPC 11150
Acacia crassicarpa
Indonesia
M.J. Wingield
GU214559, AY752163, GU214459
Passalora sequoiae
CPC 11258
Juniperus virginiana
U.S.A.
C.S. Hodges
GU214667, GU214667, GU214667
Passalora sp.
CBS 115525; CPC 3951
Tilia americana
Canada
K. Seifert
GU214560, AY293064, GU214460
CPC 12319
Ambrosia artemisifolia
var. elatior
South Korea
H.D. Shin
GU214668, GU214668, GU214668
Passalora vaginae
CBS 140.34; DSM 1148;
IMI 303641
Saccharum oficinarum
Taiwan
—
GU214561, AF222832, GU214461
Passalora zambiae
CBS 112970; CPC 1228
Eucalyptus globulus
Zambia
T. Coutinho
GU214562, AY725522, EU019272
CBS 112971; CMW
14782; CPC 1227
Eucalyptus globulus
Zambia
T. Coutinho
GU214563, AY725523, EU019273
Passalora-like genus
CPC 11876
Avicermia sp.
South Africa
W. Gams
GU214564, GU214642, GQ852622
Penidiella columbiana
CBS 486.80
Paepalanthus
columbianus
Colombia
W. Gams
GU214565, AJ244261, EU019274
Phacellium paspali
CBS 113093; RoKI 1144
Setaria palmicola
Taiwan
R. Kirschner &
C.-J. Chen
GU214669, GU214669, GU214669
Phaeophleospora atkinsonii
CBS 124565; ICMP 17860 Leaf of Hebe sp.
New Zealand
—
—, GU214643, GU214462
CBS 124566; ICMP 17862 Leaf of Hebe sp.
New Zealand
—
—, GU214644, GU214463
Phaeophleospora
eugeniicola
CPC 2557
Eugenia sp.
Brazil
—
GU214566, FJ493190, FJ493208
CPC 2558
Eugenia sp.
Brazil
—
GU214567, FJ493191, FJ493209
Phloeospora maculans
CBS 115123
—
—
C.F. Hill
GU214670, GU214670, GU214670
Piedraia hortae var. hortae
CBS 374.71
Man
French Guiana
—
—, GU214645, GU214464
CBS 375.71
Man
Brazil
—
—, GU214646, GU214465
www.studiesinmycology.org
47-S3
PhylogenetiC lineages in the Capnodiales
Table 1. (Continued).
Species
Accession number1 Host
Country
Collector
GenBank Accession numbers
18S nrDNA, 5.8S nrDNA, 28S
nrDNA
CBS 480.64; IHEM 3823;
UAMH 4341
Man, hair
Brazil
—
—, GU214647, GU214466
Piedraia hortae var.
paraguayensis
CBS 276.32; VKM F-393
—
—
—
—, GU214648, GU214467
Piedraia quintanilhae
CBS 327.63; IMI 101644
Genetta tigrina
Central African
Republic
—
—, —, GU214468
Polychaeton citri
CBS 116435
Citrus aurantium, leaf,
with Pseudococcus citri
Iran
R. Zare &
W. Gams
—, GU214649, GU214469
Pseudocercospora
angolensis
CBS 112933; CPC 4118
Citrus sp.
Zimbabwe
—
GU214568, AY260063, GU214470
CBS 149.53; ATCC 11669
Citrus sinensis
Angola
—
AY251106, AF222847, GU214471
Pseudocercospora
atromarginalis
CPC 11372
Solanum nigrum
South Korea
H.D. Shin
GU214671, GU214671, GU214671
Pseudocercospora
chengtuensis
CPC 10785
Lycium chinense
South Korea
H.D. Shin
GU214672, GU214672, GU214672
Pseudocercospora cordiana
CBS 114685; CPC 2552
Cordia goeldiana
Brazil
P.W. Crous &
R.L. Benchimol
GU214569, AF362054, GU214472
Pseudocercospora cruenta
CBS 462.75
Phaseolus sp.
Fiji
W. IJzermansLutgerhorst
AY251105, AF362065, GU214473
CPC 10846
Vigna sp.
Trinidad
H. Booker
GU214673, GU214673, GU214673
CPC 10802
Eucommia ulmoides
South Korea
H.D. Shin
GU214674, GU214674, GU214674
Pseudocercospora
eucommiae
Pseudocercospora ijiensis
X300
Musa sp.
Tonga
—
GU214570, AY752150, GU214474
Pseudocercospora fuligena
CPC 12296
Lycopersicum sp.
Thailand
—
GU214675, GU214675, GU214675
Pseudocercospora griseola
f. griseola
CBS 194.47; ATCC 22393
Phaseolus vulgaris
Portugal
—
DQ289861, DQ289801, GU214475
CBS 880.72
Phaseolus vulgaris
Netherlands
H. A. v. Kesteren
DQ289862, DQ289802, GU214476
Pseudocercospora humuli
CPC 11358
Humulus japonicus
South Korea
H.D. Shin
GU214676, GU214676, GU214676
Pseudocercospora kaki
CPC 10636
Diospyros lotus
South Korea
H.D. Shin
GU214677, GU214677, GU214677
Pseudocercospora luzardii
CPC 2556
Hancornia speciosa
Brazil
A.C. Alfenas &
P.W. Crous
GU214571, AF362057, GU214477
Pseudocercospora
macrospora
CBS 114696; CPC 2553
Bertholletia excelsa
Brazil
P.W. Crous &
R.L. Benchimol
GU214572, AF362055, GU214478
Pseudocercospora ocimicola
CPC 10283
Ocimum basilicum
Mexico
M.E. Palm
GU214678, GU214678, GU214678
Pseudocercospora opuntiae
CBS 117708; CPC 11772
Opuntia sp.
Mexico
M. De Jesus
Yanez
GU214679, GU214679, GU214679
Pseudocercospora pallida
CPC 10776
Campsis grandilora
South Korea
H.D. Shin
GU214680, GU214680, GU214680
Pseudocercospora
paraguayensis
CBS 111317; CPC 1458
Eucalyptus nitens
Brazil: Suzano
P.W. Crous
GU214573, AF309596, GU214479
Pseudocercospora
protearum var. leucadendri
CPC 1869
Leucadendron sp.
South Africa
S. Denman &
P.W. Crous
AY251107, AY260089, GU214480
Pseudocercospora
pseudoeucalyptorum
CBS 114242; CMW
14908; CPC 10390
Eucalyptus globulus
Spain
J.P.M. Vazquez
GU214574, AY725526, GU214481
Pseudocercospora punctata
CBS 113315
Syzygium cordatum
South Africa
M.J. Wingield
EU167582, EU167582, GU214407
CPC 10532
Syzygium cordatum
South Africa
M.J. Wingield
GU214659, GU214659, GU214659
CPC 11592
Zelkova serrata
South Korea
H.D. Shin
GU214575, DQ303085, GU214482
Pseudocercospora sp.
Pseudocercospora vitis
CPC 11595
Vitis vinifera
South Korea
H.D. Shin
DQ073923, DQ073923, GU214483
Pseudocercospora-like
genus
CPC 10712
Quercus sp.
Netherlands
G. Verkley
GU214681, GU214681, GU214681
Pseudocercosporella
capsellae
CPC 10301
Brassica sp.
U.K.
R. Evans
GU214662, GU214662, GU214662
Pseudocercosporella fraxini
CPC 11509
Fraxinus rhynchophylla
South Korea
H.D. Shin
GU214682, GU214682, GU214682
Pseudocercosporella sp.
CBS 112737; CPC 3959
Rhus typhina
Canada
K. Seifert
GU214684, GU214684, GU214684
CPC 4008
Rhus typhina
Canada
K. Seifert
GU214686, GU214686, GU214686
47-S4
Crous et al.
Table 1. (Continued).
Species
Accession number1 Host
Country
Collector
GenBank Accession numbers
18S nrDNA, 5.8S nrDNA, 28S
nrDNA
CPC 10050
Rubus oldhamii
South Korea
H.D. Shin
GU214685, GU214685, GU214685
CPC 11414
Vicia amurense
South Korea
H.D. Shin
GU214683, GU214683, GU214683
Pseudotaeniolina globosa
CBS 109889
Rock
Italy
C. Urzi
GU214576, AY128700, EU019283
Rachicladosporium cboliae
CBS 125424; CPC 14034
Twig debris
U.S.A.
P.W. Crous
—, GU214650, GU214484
Ramichloridium apiculatum
CPC 12310
Vicia amurensis
South Korea
H.D. Shin
GU214687, GU214687, GU214687
Ramichloridium cerophilum
CBS 103.59; MUCL
10034
Sasa sp.
Japan
—
EU041798, EU041798, GU214485
Ramichloridium musae
CBS 190.63; MUCL 9557
Musa sapientum
—
—
GU214577, EU041800, EU041857
Ramichloridium-like genus
CPC 10672
Phellodendron amurense
South Korea
H.D. Shin
GU214688, GU214688, GU214688
Ramularia acroptili
CBS 120252
Acroptilon repens
Turkey
R. Sobhian
GU214689, GU214689, GU214689
Ramularia brunnea
CPC 4903
—
—
—
GU214691, GU214691, GU214691
Ramularia coleosporii
CPC 11516
Plectranthus excisus
South Korea
H.D. Shin
GU214692, GU214692, GU214692
Ramularia endophylla
CBS 113265
Quercus robur
Netherlands
G. Verkley
AY490775, AY490763, AY490776
Ramularia grevilleana
CPC 656
Fragaria sp.
South Africa
P.W. Crous
GU214578, AF173312, GU214486
Ramularia nagornyi
CBS 120253
Centaurea solstitiales
Greece
D. Berner
GU214579, EU019257, EU019257
Ramularia pratensis var.
pratensis
CPC 11294
Rumex crispus
South Korea
H.D. Shin
GU214580, EU019284, EU019284
Ramularia sp.
CBS 324.87
leaf spot on Brassica sp.,
in Mycosphaerella sp.
Netherlands
—
GU214581, EU019285, EU019285
CPC 10066
Alangium platanilium
South Korea
H.D. Shin
GU214690, GU214690, GU214690
CPC 11297
Stellaria aquatica
South Korea
H.D. Shin
GU214693, GU214693, GU214693
Ramularia uredinicola
CPC 10813
Salix sp.
South Korea
H.D. Shin
GU214694, GU214694, GU214694
Ramularia-like genus
CPC 10852
Polygonum sp.
South Korea
H.D. Shin
GU214695, GU214695, GU214695
Ramulispora sorghi
CBS 110578; CPC 905
Sorghum sp.
South Africa
D. Nowell
AY251110, AY259131, GU214487
CBS 110579; CPC 906
Sorghum sp.
South Africa
D. Nowell
AY251111, AY259132, GU214488
Readeriella dimorphospora
CBS 120034; CPC 12636
Eucalyptus nitens
Australia
—
GU214521, EF394850, EU019258
Readeriella mirabilis
CBS 116293; CPC 10506
Eucalyptus fastigata
New Zealand
W. Gams
EU754110, AY725529, EU019291
Schizothyrium pomi
CBS 228.57
—
Italy
R. Ciferri
EF134947, EF134947, EF134947
CBS 406.61
Rubus idaeus
Netherlands
—
EF134949, EF134949, EF134949
Scorias spongiosa
CBS 486.50
Polygonum sachalinense
Netherlands
—
EF134948, EF134948, EF134948
CBS 325.33
Aphid
—
—
GU214696, GU214696, GU214696
Septoria apiicola
CBS 400.54; IMI 092628
Apium graveolens
Netherlands
J.A. von Arx
GU214584, AY152574, GU214490
Septoria convolvuli
CBS 102325
Calystegia sepium
Netherlands
G. Verkley
GU214697, GU214697, GU214697
Septoria cucubali
CBS 102368
Cucubalus baccifer
Netherlands
G. Verkley
GU214698, GU214698, GU214698
Septoria dysentericae
CPC 12328
Daucus carota
Brazil
N. Massola
GU214699, GU214699, GU214699
Septoria lactucae
CBS 352.58
Lactuca sativa
Germany
—
GU214585, AY489282, GU214491
Septoria leucanthemi
CBS 109090
Chrysanthemum
leucanthemum
Austria
G. Verkley
GU214586, AY489277, GU214492
Septoria obesa
CBS 354.58; BBA 8554;
IMI 091324
Chrysanthemum indicum
Germany
—
GU214587, AY489285, GU214493
Septoria protearum
CPC 1470
Protea cynaroides
South Africa
L. Viljoen
GU214588, AY260081, GU214494
Septoria pyricola
CBS 222.31; CPC 3677
Pyrus communis
—
—
GU214589, AY152591, GU214495
Septoria quercicola
CBS 663.94
Quercus robur
Netherlands
—
GU214590, AY490771, GU214496
Septoria rosae
CBS 355.58; ATCC
24311; PD 341; CPC 4302
Rosa sp.
—
—
AY251113, AY293065, GU214497
Septoria senecionis
CBS 102366
Senecio luviatilis
Netherlands
G. Verkley
GU214591, AY489272, GU214498
Septoria-like genus
CBS 102377
Castanea sativa
Netherlands
G. Verkley
GU214592, AY152588, GU214499
Sonderhenia eucalypticola
CPC 11252
Eucalyptus globulus
Spain
M.J. Wingield
GU214593, DQ303064, GU214500
Sphaerulina polyspora
CBS 354.29
—
—
—
—, GU214651, GU214501
Staninwardia suttonii
CBS 120061; CPC 13055
Eucalyptus robusta
Australia
B.A. Summerell
GU214594, DQ923535, DQ923535
www.studiesinmycology.org
47-S5
PhylogenetiC lineages in the Capnodiales
Table 1. (Continued).
Species
Accession number1 Host
Country
Collector
GenBank Accession numbers
18S nrDNA, 5.8S nrDNA, 28S
nrDNA
Stenella araguata
CBS 105.75; ATCC
24788; FMC 245
Man
Venezuela
—
GU214596, EU019250, EU019250
Stigmina platani
CBS 110755; IMI 136770;
CPC 4299
Platanus orientalis
India
—
GU214598, AY260090, FJ839663
Stigmina synanamorph
CPC 11721
Platanus occidentalis
South Korea
H.D. Shin
GU214700, GU214700, GU214700
Stomiopeltis betulae
CBS 114420
Betula sp.
Sweden
K. & L. Holm
GU214701, GU214701, GU214701
Teratosphaeria aff. nubilosa
CBS 114419; CPC 10497
Eucalyptus globulus
New Zealand
—
GU214599, AY725574, EU019303
CBS 116283; CPC 10495
Eucalyptus globulus
Spain
W. Gams
GU214600, AY725573, GU214503
Teratosphaeria alcornii
CBS 313.76; CPC 3632
Eucalyptus tessellaris
Australia
J.L. Alcorn
GU214514, AF362061, EU019245
Teratosphaeria angophorae
CBS 120493; DAR 77452
Angophora loribunda
Australia
A.J. Carnegie
—, GU214652, GU214504
Teratosphaeria bellula
CBS 111700; CPC 1821;
JT 196
Protea eximia
South Africa
J.E. Taylor
GU214601, EU019301, EU019301
Teratosphaeria cryptica
CBS 110975; CMW 3279;
CPC 936
Eucalyptus globulus
Australia
A.J. Carnegie
GU214602, AF309623, GU214505
Teratosphaeria destructans
CBS 111369; CPC 1366
Eucalyptus grandis
Indonesia
M.J. Wingield
GU214603, DQ267595, EU019287
CBS 111370; CPC 1368
Eucalyptus sp.
Indonesia
P.W. Crous
GU214702, GU214702, GU214702
Teratosphaeria ibrillosa
CPC 1876
Protea nitida
South Africa
J.E. Taylor
EU019282, EU019282, GU214506
Teratosphaeria juvenalis
CBS 110906; CMW
13347; CPC 40
Eucalyptus cladocalyx
South Africa
P.W. Crous
AY720715, AY725513, FJ493217
CBS 111149; CPC 23
Eucalyptus cladocalyx
South Africa
P.W. Crous
AY720714, AY725514 , EU019294
CBS 110756; CPC 1872
Protea nitida
South Africa
J.E. Taylor
GU214519, AY260095, EU019254
CBS 111029; CPC 1488
Protea sp.
South Africa
P.W. Crous
AY251118, AY260096, FJ493199
Teratosphaeria mexicana
CBS 110502; CMW 14461
Eucalyptus globulus
Australia
—
GU214604, AY725558, GU214507
CBS 120744; CPC 12349
Eucalyptus sp.
U.S.A.: Hawaii
W. Gams
GU214605, EU019302, EU019302
Teratosphaeria molleriana
CBS 111164; CMW 4940;
CPC 1214
Eucalyptus globulus
Portugal
M.J. Wingield
GU214606, AF309620, EU019292
CBS 116370; CPC 10397
Eucalyptus globulus
Spain
J.P.M. Vazquez
GU214607, AY725561, GU214508
Teratosphaeria macowanii
CPC 4577
Eucalyptus sp.
Australia
—
GU214582, AY725524, GU214489
CBS 115669; CPC 933
Eucalyptus nitens
South Africa
M.J. Wingield
GU214608, AY725548, GU214509
CBS 116005; CMW 3282;
CPC 937
Eucalyptus globulus
Australia
A.J. Carnegie
GU214609, AY725572, GU214510
CBS 112896; CMW 4937;
CPC 1004
Eucalyptus grandis
South Africa
M.J. Wingield
AY251119, AF309604, EU019305
CBS 112973; CMW 4936;
CPC 1005
Eucalyptus grandis
South Africa
M.J. Wingield
GU214610, AF309605, GU214511
Teratosphaeria
pseudosuberosa
CBS 118911; CPC 12085
Eucalyptus sp.
Uruguay
M.J. Wingield
GU214611, DQ303011, EU019256
Teratosphaeria secundaria
CBS 115608; CPC 504
Eucalyptus grandis
Brazil
A.C. Alfenas
GU214612, DQ303018, EU019306
Teratosphaeria nubilosa
Teratosphaeria ohnowa
Teratosphaeria sp.
CBS 208.94; CPC 727
Eucalyptus grandis
Indonesia
A.C. Alfenas
GU214613, AY626982, EU019307
Teratosphaeria
stellenboschiana
CBS 116428; CPC 10886
Eucalyptus sp.
South Africa
P.W. Crous
GU214583, AY725518, EU019295
Teratosphaeria suberosa
CPC 11032
Eucalyptus sp.
Colombia
M.J. Wingield
GU214614, DQ303044, GU214512
Teratosphaeria suttonii
CPC 11279
Eucalyptus tereticornis
Bolivia
M.J. Wingield
GU214615, DQ303055, FJ493222
CPC 12352
Eucalyptus sp.
U.S.A.: Hawaii
W. Gams
GU214616, EU019288, EU019288
Teratosphaeria toledana
CBS 113313; CMW 14457
Eucalyptus sp.
Spain
P.W. Crous &
G. Bills
GU214617, AY725580, GU214513
CBS 115513; CPC 10840
Eucalyptus sp.
Spain
P.W. Crous &
G. Bills
GU214618, FJ493198, FJ493225
Teratosphaeria verrucosa
CPC 18
Eucalyptus cladocalyx
South Africa
P.W. Crous
AY720713, AY725517, EU019293
Thedgonia-like genus
CPC 12304
Oplismenus
undulatifolius
South Korea
H.D. Shin
GU214703, GU214703, GU214703
47-S6
Crous et al.
Table 1. (Continued).
Species
Accession number1 Host
Country
Collector
GenBank Accession numbers
18S nrDNA, 5.8S nrDNA, 28S
nrDNA
Toxicocladosporium irritans
CBS 185.58
Mouldy paint
Suriname
M.B. ScholSchwarz
GU214619, EU040243, EU040243
Verrucisporota daviesiae
CBS 116002; VPRI 31767
Daviesia latifolia
Australia
V. Beilharz
GU214620, FJ839633, FJ839669
Verrucisporota proteacearum
CBS 116003; VPRI 31812
Grevillea sp.
Australia
J.L. Alcorn
GU214621, FJ839635, FJ839671
Zasmidium anthuriicola
CBS 118742
Anthurium sp.
Thailand
C.F. Hill
GU214595, FJ839626, FJ839662
Zasmidium citri
CBS 116366; CMW
11730; CPC 10522
Acacia mangium
Thailand
K. Pongpanich
GU214597, AY752145, GU214502
1
ATCC: American Type Culture Collection, Virginia, U.S.A.; BBA: Biologische Bundesanstalt für Land- und Forstwirtschaft, Berlin-Dahlem, Germany; CBS: Centraalbureau
voor Schimmelcultures, Utrecht, The Netherlands; CMW: Culture Collection of the Forestry and Agricultural Biotechnology Institute (FABI) of the University of Pretoria,
Pretoria, South Africa; CPC: Culture collection of Pedro Crous, housed at CBS; DAOM: Plant Research Institute, Department of Agriculture (Mycology), Ottawa, Canada;
DAR: Plant Pathology Herbarium, Orange Agricultural Institute, Forest Road, Orange. NSW 2800, Australia; DSM: Deutsche Sammlung von Mikrorrganismen und Zellkulturen
GmbH, Braunschweig, Germany; ETH: Swiss Federal Institute of Technology Culture Collection, Zurich, Switzerland; FMC: Venezuelan School of Medicine; IAM: IAM
Culture Collection, Institute of Molecular and Cellular Biosciences, The University of Tokyo, Japan; ICMP: International Collection of Micro-organisms from Plants, Landcare
Research, Private Bag 92170, Auckland, New Zealand; IFO: Institute for Fermentation, Osaka, Japan; IHEM: Collection of the Laboratorium voor Microbiologie en Microbiele
Genetica, Rijksuniversiteit, Ledeganckstraat 35, B-9000, Gent, Belgium; IMI: International Mycological Institute, CABI-Bioscience, Egham, Bakeham Lane, U.K.; IPO: Culture
collection of the Research Institute for Plant Protection, Wageningen, The Netherlands; JCM: Japan Collection of Microorganism, RIKEN BioResource Center, Japan; JT:
Working collection of Joanne E. Taylor; LSHB: London School of Hygiene & Tropical Medicine, London, U.K.; MPFN: Culture collection at the Laboratoire de Pathologie
Forestie`re, INRA, Centre de Recherches de Nancy, 54280 Champenoux, France; MUCL: Université Catholique de Louvain, Louvain-la-Neuve, Belgium; PD: Plant Protection
Service, Wageningen, The Netherlands; RoKI: Private culture collection Roland Kirschner; TNS: Herbarium of the National Museum of Nature and Science of Japan, Tokyo,
Japan; UAMH: University of Alberta Microfungus Collection and Herbarium, Edmonton, Alberta, Canada; VKM: All-Russian Collection of Microorganisms, Russian Academy
of Sciences, Institute of Biochemistry and Physiology of Microorganisms, 142292 Pushchino, Moscow Region, Russia; VPRI: Victorian Department of Primary Industries,
Knoxield, Australia; WAC: Department of Agriculture Western Australia Plant Pathogen Collection, Perth, Australia; X: Working collection of Mahdi Arzanlou.
www.studiesinmycology.org
47-S7