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Rust fungi on Annonaceae II: the genus
Dasyspora
Berk. & M.A. Curtis
Ludwig Beenken
1
ETH Zurich, Institute of Integrative Biology, 8092
Zurich, Switzerland
Stefan Zoller
ETH Zurich, Genetic Diversity Centre, 8092 Zurich,
Switzerland
Reinhard Berndt
ETH Zurich, Institute of Integrative Biology, 8092
Zurich, Switzerland
Abstract
:
Dasyspora gregaria
, the single species of the
allegedly monotypic rust genus
Dasyspora
(Basidio-
mycota, Pucciniales), was investigated by light micros-
copy and DNA sequencing (ITS1–5.8S–ITS2 region,
partial LSU and SSU of the nuclear rDNA, mt
cytochrome oxidase subunit 3). Both methods indi-
cated that
D. gregaria
is not a single species but can
be split in 11 distinct taxa, each of which appear
confined to a single
Xylopia
species (Annonaceae)
host. Herein nine of these are described as new. Both
the phylogenetic analyses and morphology show that
the species are grouped into two main clades
designated
Dasyspora gregaria
and
D. winteri
. The
first comprises
D. gregaria
, the type species of the
genus, which is restricted to
X. cayennensis,
two new
species on
X. aromatica
,
D. segregaria
from northern
South America and
D. echinata
from Brazil. The
second clade is formed by
D. winteri
, recombined
from
Puccinia winteri
on
X. sericea,
and the new
species
D. amazonica
on
X. amazonica, D. emarginatae
on
X. emarginata
,
D. frutescentis
on
X. frutescens
,
D.
ferrugineae
on
X. frutescens
var.
ferruginea
,
D. guia-
nensis
on
X. benthamii
,
D. mesoamericana
on
X.
frutescens
, and
D. nitidae
on
X. nitida
.
Dasyspora
frutescentis
and
D. mesoamericana
were not clearly
distinguishable by their morphology and host associ-
ations but differed from another in their sequences
and geographic distributions. They are considered
cryptic species. An identification key and the distri-
butions are given for all recognized species. Along
with molecular data we discuss the systematic position
of
Dasyspora
in the Pucciniales.
Key words:
cryptic species, mt cytochrome oxidase
subunit 3, nuclear-rDNA, Pucciniales/Uredinales, rust
fungal specific primers, Uropyxidaceae,
Xylopia
INTRODUCTION
Berkeley and Curtis (1853) proposed
Dasyspora
as a
new genus of Pucciniales (5Uredinales, Kirk et al.
2008) based on
D. foveolata
Berk. & Curt., a
microcyclic rust known only from
Xylopia
L. (Anno-
naceae) species in the Neotropics.
Dasyspora
is
characterized by verrucose, two-celled,
Puccinia
-like
teliospores. Each teliospore cell has a single germ
pore that is surrounded by crowded, cylindrical and
apically branching projections. The genus differs
from
Puccinia
in its spermogonial type (Group VI,
type 5 vs. Group V, type 4 in
Puccinia
; Cummins and
Hiratsuka 2003) and therefore was assigned to
Uropyxidaceae (Arthur) Cummins and Y. Hirats. by
Cummins and Hiratsuka (1983, 2003).
Dasyspora
foveolata
generally has been synonymized with the
older name
Puccinia gregaria
Kunze, which later was
recombined into
D. gregaria
(Kunze) Henn.
Puccinia
winteri
Pazschke was considered to be a further
synonym of
D. gregaria
(Hennings 1895, 1896;
Saccardo 1895; Trotter and Cash 1972; Magnus
1892; Arthur 1927; Cummins and Hiratsuka 2003).
Therefore
Dasyspora
has been considered a monotyp-
ic genus distributed throughout the tropical Americas
(Arthur 1927, 1929; Buritica´ and Pardo Cardona
1996; Dennis 1970; Hennings 1895; Hennen and
Figueiredo 1981; Hennen et al. 2005, Jørstad 1956;
Mains 1935, 1939; Sydow 1925; Vie´gas 1945). The host
genus
Xylopia
(Annonaceae) consists of 100–160
species and is pantropically distributed (Kessler
1993), but
Dasyspora
until now was known only from
a few of the ca. 50 Neotropical
Xylopia
species:
X.
aromatica
(Lam.) Mart. (5
X. grandiflora
A. St.Hil.),
X. frutescens
Aubl.,
X. ochranta
Mart., and several
undertermined
Xylopia
spp. (Hennen et al. 2005).
In a monographic study on rust fungi of Annonaceae
(Beenken and Berndt 2010) we investigated numerous
specimens of
Dasyspora
from a broad geographic range
and different host species. By analyzing micromorpho-
logical characters and nuclear and mitochondrial gene
sequences (ITS1–5.8S–ITS2, LSU, and SSU of n-rDNA,
mt cytochrome oxidase subunit 3) we investigated
the phylogenetics and distribution of
Dasyspora
and
describe nine new species.
MATERIALS AND METHODS
Fungal collection and light microscopy.—
Rust infected leaves
of several
Xylopia
species were collected in French Guiana
in 2009 (FIG. 1a). Small pieces of leaves bearing telia
(FIG. 1b, on the left) were cut out and dried for DNA
Submitted 23 Feb 2011; accepted for publication 25 Oct 2011.
1
Corresponding author: ludwig.beenken@env.ethz.ch
Mycologia,
104(3), 2012, pp. 659–681. DOI: 10.3852/11-068
#2012 by The Mycological Society of America, Lawrence, KS 66044-8897
659
extraction with silica gel. Host plants and fungal material to
be used for morphological investigation were dried between
blotting paper in a plant press at room temperature or on a
fungus drying oven. Host plants were identified with the
works of Fries (1931, 1939), Steyermark et al. (1995), Maas
and Maas-van de Kamer (2002) and Funk et al. (2007),
comparing with scans of type specimens in (http://plants.
jstor.org) and comparing with specimens in the herbaria
Cayenne (CAY), Munich (M) and Berlin (B). Some species
were determined by David M. Johnson (Ohio Wesleyan
University, Delaware, Ohio), a specialist of
Xylopia
.
Herbarium specimens were borrowed from B, BPI, BRUX,
M, NY, PMA, PUR, S and Z +ZT (acronyms according to
Index Herbariorum, Thiers 2011). Infected leaves were taken
from plant herbarium sheets of
Xylopia
spp. in M and B to
create four new fungal specimens. The investigated speci-
mens are listed under the respective fungal taxa.
Micromorphology was examined by light microscopy.
Spore scrapes and hand sections (FIG. 2) were mounted in
lactophenol or in lactophenol with cotton blue. A camera
lucida attached to a Zeiss Axioskop was used to make
drawings. Digital images were taken with a Canon Power
FIG.1.
Dasyspora gregaria
on
Xylopia cayennensis
(French Guiana, ZT Myc 3393). a. Collection in the field. Rust fungal
infections have caused yellow spots on the upper leaf sides. b. Two groups of telia in bleached spots on the lower leaf surface.
On the right, the telia are overgrown by the orange, cottony sporangiophores of
Stomatochroon
sp. This aerial alga of the order
Trentepohliales is very often associated with
Dasyspora
spp. and was misinterpreted as aecia or uredinia of
Dasyspora
by several
authors in the past (Hennen and Figueiredo 1981).
FIG.2.
Dasyspora frutescentis
on
X. frutescens
var.
ferruginea
(HeRB A-141), leaf sections. a. Telium. b. Spermogonium.
660 MYCOLOGIA
Shot A640 digital camera through the same microscope.
Measurements of 30–50 spores comprise the range and
arithmetic means. Spore measurements do not include the
spore ornamentation.
Terminology follows Cummins and Hiratsuka (2003). To
illustrate the geographical distribution of each species Flora
Neotropica base map No. 1 (prepared by Hendrik R.
Rypkema, Department of Systematic Botany, State Univer-
sity of Utrecht, the Netherlands 1989) was scanned and
modified with standard graphics software (Adobe InDesign
CS4, 6.0.6) (FIG. 3).
DNA extraction.—
Pieces (ca. 10–20 mm
2
) with visible fungal
infection were excised from dry host leaves and finely
ground with a Retsch mixer mill. DNA was extracted with
NucleoSpin Plant II extraction kit (Macherey-Nagel, Du¨ren,
Germany), following the manufacturer’s standard protocol
for plant tissue, which was modified by eluting DNA in 40 mL
elution buffer. DNA extractions were numbered consecu-
tively (TABLE I).
PCR amplification and primer design.—
Fungal nuclear
rDNA was amplified directly from extracted DNA (1 mL
per 20 mL reaction volume) by polymerase chain reaction
(PCR) with Phusion High-Fidelity DNA Polymerase (Finn-
zymes,Espoo,Finland)accordingthemanufacturer’s
pipetting and cycling instructions (35 cycles, annealing
temperatures see below). The internal transcribed spacer
region (ITS1–5.8S–ITS2), partial LSU (28S) and SSU (18S)
of the nuclear rDNA repeat and the mitochondrial
cytochrome oxidase subunit 3 (CO3) were amplified. If it
was impossible to get the DNA fragments in one PCR
reaction, they were amplified in several overlapping
fragments using additional internal primers (TABLE II,
SUPPLEMENTARY FIG. 1). This was necessary customarily when
herbarium specimens were used and let us amplify DNA
from specimens of up to 123 y old. To prevent amplification
of the DNA of host plants or of possible fungal contami-
nants rust fungal specific primers were used. For this
purpose additional specific primers (TABLE II, SUPPLEMEN-
TARY FIG. 1) were designed based on alignments of
Dasyspora
sequences, which were amplified and sequenced
with primers described in literature (TABLE II), and
analogous sequences of rust fungi from GenBank. ITS and
LSU primers were tested for specificity against sequences of
the rust parasitic
Eudarluca caricis
(GenBank AY836372),
which was found on our samples, as well as against
sequences of potential fungal contaminations like
Ramulis-
pora
spp. (GenBank GU214488),
Rhodotorula
spp. (Gen-
Bank GU080059) and
Candida
spp. (GenBank AB495288).
The primers for the SSU were checked against a sequence
of
Annona muricata
L. (GenBank AF206850) because PCRs
with the SSU primers NS1, NS3, NS4, NS5 and NS6 (White
et al. 1990) also amplified plant DNA.
The following primer combinations were used (annealing
temperatures in brackets).
ITS1–5
.
8S–ITS2
: ITS5-u/ITS4-u
(53 C); ITS5-u/ITS4rust (53 C); ITS5-u/Rust2 (52 C) and
Rust2inv/ITS4rust (52 C).
LSU
: Rust2inv/LR6 or LRust1R/
LR6 (52 C); LRust1R/LRust3 (50 C) and LRust3R/LR6
(50 C).
SSU
: NS1/Rust18SR (55 C); NS1/NSrust3R (59 C)
and NSrust2/Rust18SR (59 C); NS1/NSrust1R (57 C),
NSrust1/NSrust2R (54 C), NS3/NSrust3R (63 C),
NSrust2/NSrust5R (57 C), NS5/NSrust7R (54 C) and
NSrust6/Rust18SR (57 C).
CO3
: CO3_F1/CO3_R1 (54 C);
CO3_F1/CO3rust3R (55 C) and CO3rust2F/CO3_R1
(57 C); CO3_F1/CO3rust4R (55 C) and CO3rust2F/
CO3_R1 (57 C). PCR products were purified enzymatically
(http://www.nucleics.com/DNA_sequencing_support/
exonucleaseI-SAP-PCR-protocol.html).
Sequencing.—
Cycle sequencing reactions were performed
with the Big Dye Terminator kit 3.1 (Applied Biosystems,
FIG. 3. Collections sites of
Dasyspora
illustrating the known distribution of each species. a.
Dasyspora gregaria
clade:
triangles 5
D. segregaria
; circles 5
D. gregaria
; stars 5
D. echinata
.b.
Dasyspora winteri
clade: diamonds 5
D. mesoamericana
;
squares 5
D. frutescentis
;A5
D. amazonica
;E5
D. emarginatae
;F5
D. ferrugineae
;G5
D. guianensis
;N5
D. nitidae
;W5
D. winteri
.
BEENKEN ET AL:
D
ASYSPORA
661
TABLE I. Specimens and GenBank accession numbers used in the phylogenetic analyses of
Dasyspora
Species Host
a
Country Voucher Isolate ITS LSU SSU CO3
D. amazonica X
. cf.
amazonica
Brazil BPI US0116382 58 JF263460 JF263496 JF263512
D. echinata X. aromatica
Brazil ZT HeRB 8486 05 JF263461 — — —
D. echinata X. aromatica
Brazil BPI 746651 66 JF263462 JF263497 JF263513
D. echinata X. aromatica
Brazil M 0148197 76 JF263464 — — —
D. emarginatae X. emarginata
Brazil PUR N6196 29 JF263465 JF263498 JF263514
D. ferrugineae X. frutescens
var.
ferruginea
French Guiana ZT Myc 3407 08 JF263466 — —
D. ferrugineae X. frutescens
var.
ferruginea
French Guiana ZT Myc 3404 21 JF263467 JF263499 JF263515
D. frutescentis X. frutescens
var.
frutescens
French Guiana ZT Myc 3403 02 JF263468 JF263500 JF263516
D. frutescentis X. frutescens
var.
frutescens
French Guiana ZT Myc 3401 06 JF263469 — — —
D. frutescentis X. frutescens
var.
frutescens
Brazil ZT HeRB 8530 15 JF263470 — — —
D. frutescentis X. frutescens
var.
ferruginea
Venezuela ZT HeRB A-141 27 JF263471 JF263501 JF263517
D. frutescentis X. frutescens
var.
frutescens
Guyana BPI US0116392 69 JF263472 — —
D. frutescentis X. frutescens
var.
frutescens
Venezuela BPI 853918 70 JF263473 — — —
D. gregaria X. cayennensis
French Guiana ZT Myc 3393 03 JF263474 — —
D. gregaria X. cayennensis
French Guiana ZT Myc 3396 09 JF263475 — — —
D. gregaria X. cayennensis
French Guiana ZT Myc 3394 17 JF263476 — — —
D. gregaria X. cayennensis
French Guiana ZT Myc 3397 18 JF263477 JF263502 JF263518
D. gregaria X. cayennensis
French Guiana ZT Myc 3395 19 JF263478 — — —
D. guianensis X. benthamii
French Guiana ZT Myc 3413 20 JF263479 JF263503 JF263519
D. mesoamericana X. frutescens
var.
frutescens
Panama PUR 42390 59 JF263480 JF263504 JF263520
D. mesoamericana X. frutescens
var.
frutescens
Belize BPI US0116395 60 JF263481 — — —
D. mesoamericana X. frutescens
var.
frutescens
Guatemala BPI US0116379 62 JF263482 — —
D. mesoamericana X. frutescens
var.
frutescens
Mexico PUR 64451 72 JF263483 — —
D. nitidae X. nitida
French Guiana ZT Myc 3409 01 JF263484 JF263505 JF263521
D. nitidae X. nitida
French Guiana ZT Myc 3408 07 JF263485 — — —
D. nitidae X. nitida
French Guiana ZT Myc 3411 11 JF263486 — — —
D. nitidae X. nitida
French Guiana ZT Myc 3412 22 JF263487 JF263506 JF263522
D. segregaria X. aromatica
Panama PMA MP4941 57 JF263488 JF263507 JF263523
D. segregaria X. aromatica
Venezuela BPI 853914 63 JF263489 — — —
D. segregaria X. aromatica
Trinidad and Tobago BPI 853915 64 JF263490 — —
D. segregaria X. aromatica
Trinidad and Tobago BPI 853916 75 JF263491 — — —
D. winteri X. sericea
Brazil S F30078 37 JF263492 JF263508 JF263524
Diorchidium
polyalthiae Polyalthia longifolia
India ZT HeRB 251
16
JF263493 JF263509 JF263525
Porotenus biporus Memora flavida
French Guiana ZT Myc 3414 79 — JF263490 JF263510 —
Puccinia popowiae Monanthotaxis caffra
South Africa ZT Myc 1976 39 JF263495 JF263511 JF263526
a
Host plants belong to the Annonaceae except for
Memora flavida
of Bignoniaceae.
662 MYCOLOGIA
Warrington, UK) according to the manufacturer’s protocol
with the same primers as used in the PCR reactions
(TABLE II). After cycle sequencing products were cleaned
by Sephadex G-50 Superfine gel filtration (Amersham) on
MultiScreen TM-HV membrane plates (Millipore, Bedford,
Massachusetts), according to the manufacturer’s instruc-
tions. Fragments were sequenced on an Applied Biosystems
3130xl Genetic Analyzer capillary sequencer. If available the
ITS region was sequenced for more than one isolate per
morphotype, host and region respectively. Preliminary tests
(not presented here) have shown that samples with
identical ITS sequences also have identical sequences of
LSU, SSU and CO3. Therefore we reduced numbers of
sequences of LSU, SSU and CO3 by selecting one
representative sample per group showing identical ITS
sequences. DNA sequences were deposited in GenBank
(TABLE I).
Alignment.—
The sequences were assembled and aligned
with Sequencher 4.8 (Gene Codes Corp., Ann Arbor,
Michigan) and MacClade 4.06 (Maddison and Maddison
2003). To improve the alignments of the ITS sequences we
used the program T-Coffee 8.91 (Notredame et al. 2000)
with default settings. Ambiguously aligned regions were
delimited following Lutzoni et al. (2000) and excluded
from phylogenetic analyses. Three datasets with a total of six
submatrices were created: (i) an ITS dataset consisting of 35
taxa and 623 nucleotide sites; (ii) a combined dataset
consisting of 15 taxa and four sub-matrices (SSU with 1630
sites, CO3 with 586 sites, LSU with 968 sites, ITS with 424
sites); (iii) a SSU-LSU dataset consisting of 43 taxa (most of
them derived from Aime 2006) and two submatrices (SSU
with 1641 sites, LSU with 808 sites); accession numbers of
sequences taken from GenBank are provided (FIG. 4). DNA
sequence alignments were deposited in TreeBASE (ID
11261).
Phylogenetic analysis.—
The aligned datasets and sub-matri-
ces were analyzed with maximum likelihood methods as
implemented in RAxML 7.2.6 (Stamatakis 2006). For the
TABLE II. Rust fungal primers
Primer Sequence T
a
Reference
ITS1–5.8S–ITS2:
ITS4rust (59-CAGATTACAAATTTGGGCT-39)48.8
ITS4-u (59-GGCTTTTCCCTCTTCAT-39)50.0 Pfunder et al. 2001
ITS5-u (59-CAAGGTTTCTGTAGGTG-39)50.0 Pfunder et al. 2001
Rust2 (59-TTTCACTGTGTTCTTCATC-39)48.8 Kropp et al. 1997
Rust2inv (59-GATGAAGAACACAGTGAAA-39)48.8 Aime 2006
LSU:
LR6 (59-CGCCAGTTCTGCTTACC-39)54.0 Vilgalys and Hester 1990
LRust1R (59-TAAGACCTCAAATCAGGT-39)48.1
LRust2 (59-TCCCTCACGGTACTTGTTTGCT-39)57.7
LRust2R (59-AGCAAACAAGTACCGTGAGGGA-39)57.7
LRust3 (59-GGGTCATTTAAAGCTAT-39)46.0
LRust3R (59-ATAGCTTTAAATGACCC-39)46.0
SSU:
NS1 (59-GTAGTCATATGCTTGTCTC-39)50.8 White et al. 1990
NS3 (59-GCAAGTCTGGTGCCAGCAGCC-39)63.6 White et al. 1990
NS5 (59-AACTTAAAGGAATTGACGGAAG-39)51.9 White et al. 1990
NSrust1 (59-GATGCTTCATTCAAATATCTG-39)49.6
NSrust1R (59-GCAATCCTTAGATTTGAGGGC-39)55.8
NSrust2 (59-TCAGTTGGGGGCATTTGTATTACA-39)56.3
NSrust2R (59-CCTGCTTTGAACACTCTAATT-39)51.7
NSrust3R (59-ATCCTTGTCTGATCCCCAGTC-39)57.4
NSrust5R (59-AGCTATCAATCTGTCAATCCT-39)51.7
NSrust6 (59-TAGTTGATGGAAGTTGGAGGC-39)55.8
NSrust7R (59-GAAGAGCAATAATTGCAATGC-39)51.7
Rust18SR (59-ACCTTGTTACGACTTTTACTTC-39)51.9 Aime 2006
CO3:
CO3_F1 (59-TCAGTATGTTATTTTAACGATGTAG-39)51.4 Vialle et al. 2009
CO3_R1 (59-TCCTCATCAGTAAACACTAATA-39)50.3 Vialle et al. 2009
CO3rust2F (59-CTGCTTCTAACATCAGCAAGC-39)55.8
CO3rust3R (59-CTATTCCCATTAATTATTCG-39)47.2
CO3rust4R (59-GGGTGCTTCAATATACTCTAG-39)53.7
a
Melting point of the primer in C, according the manufacturer’s certificate.
BEENKEN ET AL:
D
ASYSPORA
663
SSU-LSU dataset
Caeoma torreyae
was selected as outgroup
(cf. Aime 2006). For the ITS dataset the combined dataset
and its submatrices
Diorchidium polyalthiae
and
Puccinia
popowiae
were used as outgroup species because they appear
as sister group to
Dasyspora
in the SSU-LSU phylogeny
(FIG. 4). All analyses were performed assuming a general
time reversible model of nucleotide substitution (GTR),
estimating a discrete gamma distribution (GTRGAMMA option
in RAxML). One thousand rapid hill-climbing runs with
distinct starting trees were completed for each dataset.
Maximum likelihood bootstrap analyses with 1000 replicates
were performed on the individual submatrices to test for
potential conflict among the genes. Because no conflict was
found (i.e. no well supported differences in the topology)
we ran additional analyses on the combined datasets. Again
aGTR with gamma model was used but with partitions
according to the submatrices, allowing for multiple models
of substitution.
Bayesian analysis was performed with MrBayes 3.1.2
(Huelsenbeck and Ronquist 2001, Ronquist and Huelsen-
beck 2005) on the same datasets as the maximum likelihood
analysis. Independent general-time-reversible models (GTR)
with gamma distribution approximated by four categories
were implemented for all data partitions. Three indepen-
dent Bayesian runs were conducted for every dataset, each
with four chains and 1 000 000 generations, sampling every
100th tree. Post burn-in trees were collected and the
summarizations calculated only when the standard devia-
tion of split frequencies had reached below 0.01. To further
ensure that the runs reached stationarity and converged on
the same ln-likelihood scores, the resulting likelihoods, tree
topologies and model estimates were examined and
compared by eye. Posterior probability values equal to or
greater than 0.95 were considered significant. Lower
posterior probability values would be given if the corre-
sponding ML bootstrap values are higher than 50%.
Phylogenetic trees were visualized with the program
Dendroscope (Huson et al. 2007).
RESULTS
Relationships within Dasyspora.—
Eleven species of
Dasyspora
are distinguishable by combining morpho-
logical characters, molecular data and host prefer-
ences. Nine of these are described below as new
species. A total of nine
Xylopia
taxa were identified as
hosts.
The resulting maximum likelihood and Bayesian
analyses produced trees with congruent topologies
(FIG. 4 shows a tree from combined LSU and SSU
datasets; FIG. 5 shows a ML analysis from ITS1-5.8S-
ITS2 sequence data; FIG. 6 shows a tree from
combined ITS, LSU, SSU and CO3 datasets, trees
from single datasets not shown). As expected the
supporting bootstrap values and posterior probabili-
ties are higher in the trees from combined datasets
than in the ITS tree. In all phylogenetic analyses
Dasyspora
split into two well supported clades
(FIGS. 4–6), herein referred to as the
D. gregaria
and
D. winteri
clades. Within single clades resolution is
low. Only in the
D. winteri
clade are single subclades
well supported.
The same two main clades are distinguished
morphologically by the lateral ornamentation of the
teliospores (FIG. 6); high hemispherical or conical
warts were found in the
D. gregaria
clade (FIGS. 7, 8),
whereas in the
D. winteri
clade teliospores were
laterally areolate by flattened, more or less polygonal
warts, which become larger and elongated near the
spore septum (FIGS. 7, 9, 10). Within these clades the
teliospores are very similar in size and ornamentation.
Nevertheless the ornamentation of the teliospores
appeared to be the primary morphological feature to
differentiate species within the genus. The ornamen-
tation consists of long, rod-like, apically branching
projections around the poles of the spore and much
less conspicuous warts between the poles (FIG. 7). The
projections may differ in length and width. The lateral
warts are hemispherical, conical or flattened differing
in form and dimension (FIGS. 7–10, TABLE III). The
differences among size, form and arrangement of
projections and warts are small, especially in the
D.
winteri
clade, but are constant within collections from
one host taxon and serve to define distinct species
precisely. In contrast the dimensions of the teliospores
varied considerably within single collections and
between specimens of one species (TABLE III). There-
fore teliospore measurements are of negligible value
for delimitation of species due to overlapping ranges.
One exception is the collection from
X. amazonica
,
which has the smallest teliospores in the genus
Dasyspora
(TABLE III). Peripheral paraphyses were
found in the telia of all recognized
Dasyspora
species
and are reported here for the first time (FIG. 7). They
are variable in abundance, dimensions and wall
thickness within a species but do not show any
significant differences between the species.
The Dasyspora gregaria clade.—
This clade includes
D. gregaria
on
X. cayennensis
and two newly described
species occurring on
X. aromatica
:
D. echinata
and
D.
segregaria
.
Dasyspora segregaria
and
D. gregaria
differ
in LSU and ITS2 sequences but have morphologically
identical teliospores with hemispherical warts.
Dasy-
spora gregaria
and
D. echinata
are identical in the
investigated sequences (FIGS. 4–6). Only one sample
of
D. gregaria
(ZT Myc 3393) and one of
D. echinata
(BPI 746651) differ in one or two base pairs
respectively from the common ITS sequence
(FIG. 5). Nevertheless
D. echinata
differs by having
teliospores with strictly conical lateral warts and polar
projections that are shorter than in
D. gregaria
and
D. segregaria
(TABLE III; FIGS. 7, 8). The geographic
664 MYCOLOGIA
FIG. 4. Position of
Dasyspora
within Pucciniales shown by maximum likelihood analysis with RAxML 7.2.6 recovered from
combined LSU and SSU sequence data. Numbers at nodes indicate RAxML bootstrap support .50%/Bayesian posterior
probabilities .0.95. Black bars: clades including type genus/species of the family. Bold text: members of the Uropyxidaceae.
GenBank accession numbers of LSU/SSU in brackets, of new sequences in TABLE I.
BEENKEN ET AL:
D
ASYSPORA
665
distributions of the three species do not overlap
according to present knowledge (FIG.3a).
Dasyspora
echinata
occurs only in the cerrados of Brazil
(FIG. 3a), the main habitat of its host,
X. aromatica
(Castro et al. 1999).
Dasyspora segregaria
also occurs
on
X. aromatica
but is found only in northwestern
South America and Central America (FIG.3a). It
could be observed however that the hosts differed in
their leaf indumentum.
Dasyspora echinata
was
found exclusively on specimens of
X. aromatica
whose abaxial leaf surfaces were tomentose by bent
hairs. In contrast
D. segregaria
was only found on
specimens of
X. aromatica
with abaxial leaf surfaces
strigose by straight, appressed hairs.
Dasyspora
gregaria
is limited to
X. cayennensis
, a species closely
related to
X. aromatica
(Maas et al. 1993), and
distributed in the northeast of South America
(FIG. 3a). Although it was impossible to separate
D.
echinata
from
D. gregaria
by DNA sequences, it is
described here as a separate species because of
distinct differences in morphology and the different
host range.
Dasyspora segregaria
is well separated
from
D. gregaria
by the molecular data and hosts and
differs morphologically from
D. echinata
.
FIG. 5. Phylogeny of
Dasyspora
shown by maximum likelihood analysis with RAxML 7.2.6 recovered from ITS1-5.8S-ITS2
sequence data. Numbers at nodes indicate RAxML bootstrap support .50%/Bayesian posterior probabilities .0.80.
Vouchers along with their geographical origin. Species related to host preferences. GenBank accession numbers are provided
(TABLE I).
666 MYCOLOGIA
The Dasyspora winteri clade.—Puccinia winteri
on
X.
sericea
was identified as a distinct species belonging to
the genus
Dasyspora
. It is not a synonym of
D. gregaria
as Hennings (1895) suggested. Seven additional species
were distinguishable by morphology and molecular
data in this clade. They are
D. amazonica
on
X.
amazonica
,
D. guianensis
on
X. benthamii
,
D. mesoamer-
icana
on
X. frutescens
,
D. emarginatae
on
X. emarginata
,
D. frutescentis
on
X. frutescens
,
D. ferrugineae
on
X.
frutescens
var.
ferruginea
and
D. nitidae
on
X. nitida
.
Dasyspora winteri
differs from all the newly recognized
species by having the thinnest projections on its
teliospores and in its host,
X. sericea
(TABLE III,
FIG. 10). Specimens of
Dasyspora
collected on
X.
frutescens
var.
frutescens
were morphologically and
ecologically indistinguishable but belonged to two
phylogenetic lineages (FIGS. 4–6). They are regarded
as cryptic species and described below as
D. mesoamer-
icana
and
D. frutescentis
. Both are morphologically very
similar to
D. winteri
, but the polar projections of their
teliospores are thicker and the lateral warts are smaller
and often equatorially elongated (TABLE III, FIGS.9,
10). Most of the studied collections of
D. mesoamericana
had larger teliospores than
D. frutescentis
,butsome
samples had spore dimensions within the range of
D.
frutescentis
. Therefore both species cannot be distin-
guished clearly by spore measurements. In contrast to
D. frutescentis
, which is known only from northern
South America,
D. mesoamericana
appears to be
restricted Central America and environs, found in
collections from southern Mexico as northern limit of
distribution to Panama in the south (FIG.3b).
The collections of
Dasyspora
, which were found on
X. frutescens
var.
ferruginea
in French Guiana, differed
morphologically from
D. frutescentis
and all other
species of the
D. winteri
clade in having more deeply
pigmented teliospores with more densely spaced and
taller lateral warts (TABLE III; FIGS. 9, 10). Therefore
it is recognized here as the distinct species
D.
ferrugineae
.
Dasyspora ferrugineae
and
D. frutescentis
formed a well supported subclade in the molecular
analyses. Both species could be distinguished by their
ITS sequences (FIG. 5), but they share identical CO3
sequences and are not well separated by LSU and SSU
sequences (single trees not shown, FIG.4). The
specimen on
X. frutescens
var.
ferruginea
from
Venezuela (RB A-141) belongs morphologically and
genetically to
D. frutescentis
(FIGS. 5, 6). The telio-
spores of a small sample (M 0148196) on
X. discreta
,
which is closely related to
X. frutescens
(Fries 1931),
look identical to
D. frutescentis
and might belong to
this species. Sequence data were not assessed for this
specimen.
In all phylogenetic analyses
D. emarginatae
ap-
peared in a sister relationship to the remaining
species (FIGS. 4–6). The lack of discolored leaf spots
is a unique character of
D. emarginatae
that might be
FIG. 6. Phylogeny of
Dasyspora
shown by maximum
likelihood analysis with RAxML 7.2.6 recovered from
combined ITS1-5.8S-ITS2, LSU, SSU and CO3 sequence
data. Numbers at nodes indicate RAxML bootstrap support
.50%/Bayesian posterior probabilities .0.90. Species
related to lateral ornaments of teliospores in profile
(schematic drawings on the right). GenBank accession
numbers are provided (TABLE I).
FIG. 7. Teliospores and paraphyses in
Dasyspora,
ornaments in surface view and in optical section (drawings from
type collections).
BEENKEN ET AL:
D
ASYSPORA
667
due to the leathery leaves of its host
X. emarginata
.
The teliospores show the ornament typical for the
D.
winteri
clade, but the warts near the septum are not
much larger than the lateral warts (FIGS. 9e, 10). It
differs from the similar
D. amazonica
in having larger
teliospores (TABLE III). Collections of
D. nitidae
on
X.
nitida
clustered together but formed two subgroups
due to base differences in the ITS and LSU regions
(FIGS. 5, 6). These subgroups were not mirrored by
micromorphology, which was identical in all speci-
mens.
D. nitidae
teliospores differ from
D. winteri
by
their larger lateral warts and thicker projections and
are distinguishable from
D. emarginatae
, which has a
similar lateral ornamentation, by having longer pro-
jections (TABLE III; FIGS. 9, 10). However the host
plants of the two subgroups differ in leaf indumentum.
The collections of
D. nitidae
from Montagne de Kaw
(ZT Myc 3410–3412), of which one was chosen as type,
occur on
X. nitida
bearing white hairs on the abaxial
side. In contrast the
D. nitidae
collections from
Montagne Chapeau (ZT Myc 3408) and Foret de
Tiponni (ZT Myc 3409) occur on
X
.cf.
nitida
leaves
that are covered by brownish hairs.
Dasyspora amazo-
nica
and
D. guianensis
are known only from their type
collections but are distinct morphologically and in
sequences within the clade.
Dasyspora amazonica
differs from all other species in having smaller
teliospores (TABLE III). The teliospores of
D. guianen-
sis
are characterized by the shortest and thickest polar
projections of all species (TABLE III; FIGS. 9b, 10).
Position of Dasyspora within Pucciniales (F
IG
.4).—
Morphology and phylogenetic analyses identify
Dasy-
spora
as a well supported monophyletic clade. The ML
analysis as well as the Bayesian analysis of partial
nuclear LSU and SSU rDNA showed identical tree
topologies and support the
Dasyspora
clade with
100%bootstrap value and 1.00 posterior probability
respectively (FIG. 4).
Puccinia popowiae
Cooke and
Diorchidium polyalthiae
Syd. & P. Syd. occur as sister
group (100%bootstrap value/1.00 posterior proba-
bility).
Dasyspora
does not group with other investi-
gated members of Uropyxidaceae,
Tranzschelia dis-
color
(Fuckel) Tranzschel & Litv. from Asia and
Porotenus biporus
Hennen & Sota˜o from South
America and could not be assigned to any other
family of Pucciniales.
TAXONOMY
Dasyspora Berk. & M.A. Curtis, J. Acad. Nat. Sci.
Philad., N.S. 2:281 (1853)
;
Sartvellia
Berk. Introduction to Cryptogamic Botany,
London: 317, 318, fig. 72 a (1857), illegitimate name.
Not
Dasyspora
ss. Arthur (1906).
Type species:
Dasyspora foveolata
Berk. & M.A. Curtis, J.
Acad. Nat. Sci. Philad., N.S. 2: 281 (1853). Current name:
Dasyspora gregaria
(Kunze) Henn.
Species of the
Dasyspora gregaria
clade.
1. Dasyspora gregaria (Kunze) Henn., Hedwigia
35:231 (1896) FIGS.1,7,8a
Basionym:
Puccinia gregaria
Kunze, in sched. in Weigelt,
Surinam. Exsic. 1827.
FIG. 8. Teliospores in
Dasyspora
gregaria clade (surface
view and optical section). a.
D. gregaria
.b.
D. segregaria
.c.
D. echinata
. Pictures from type collections.
668 MYCOLOGIA
FIG. 9. Teliospores in
Dasyspora winteri
clade (surface view and optical section). a.
D. winteri
.b.
D. frutescentis
.c.
D.
ferrugineae
.d.
D. mesoamericana
.e.
D. emarginatae
.f.
D. nitidae
.g.
D. guianensis
.h.
D. amazonica
. Pictures from
type collections.
BEENKEN ET AL:
D
ASYSPORA
669
;
Dasyspora foveolata
Berk. & M.A. Curtis, J. Acad. Nat.
Sci. Philad., N.S. 2:281 (1853)
;
Sartvellia foveolata
(Berk. & M.A. Curtis) Berk.
Introduction to Cryptogamic Botany, London: 318,
fig. 72 a (1857), illegitimate name.
5
Puccinia compacta
Kunze ss. Thuemen, Flora 58:364–
365 (1875), not
P. compacta
Kunze ss. Kunze in Weigelt
1827 (Buba´k 1903, Sydow and Sydow 1904: 334–335, 885)
Not
Puccinia winteri
Pazschke, Hedwigia 29 (3):158–159
(1890)
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial on leaves,
subepidermal, erumpent, dark brown, densely aggre-
gated on bleached, yellowish leaf spots, 2–10 mm
diam, arranged in concentric circles or scattered.
Paraphyses peripheral, rarely between teliospores,
cylindrical to slightly clavate, 20–40 34–6 mm; wall
0.3–2 mm thick. Teliospores two-celled by transverse
septum, ellipsoid, constricted at the septum, 30–44 3
21–30 mm (mean 37.1 327.2 mm), cells subglobose;
germ pore apical in the distal cell, next to the hilum
in the proximal cell; spore wall brown, two-layered,
(1)2–3(4) mm thick, verrucose; apically and basally
with rod-like projections branched at the tip, up to 10
31.5–2 mm; lateral warts hemispherical, 1–2 mm diam
and 1–2 mm high; pedicel colorless, fragile, breaking
off close to the hilum.
Host and distribution.
On
Xylopia cayennensis
Maas
(5
X. nitida
var.
longifolia
Sagot, 5
X. longifolia
(Sagot) R.E. Fr.); Surinam, French Guiana, northern
Brazil (State of Amapa´) (FIG.3a).
LECTOTYPE designated here: SURINAM. On
Xylopia cayennensis
(det. D. M. Johnson), leg. &
exsicc. Weigelt 1827, determ. Kunze (B 70 0014139).
ISOTYPES examined: B 70 0014137, B 70 0014138,
NY 00035276 ex Herb. H. Sydow, BPI US0071773 ex
Herb. Buba` k, S F102339 ex Herb. Martius, PUR F8699
ex Herb. Schweinitz (ISOTYPE of
Dasyspora foveolata
Berk. & M.A. Curtis). Probably also ISOTYPE of
D.
gregaria
: SURINAM. (sub
Sartvellia foveolata
) (NY
00170655 ex Herb. George Massee).
Additional specimen examined:
BRAZIL. AMAPA
´: Macapa´,
Parque Zoobotanico-Macapa´, 10 Jun 1987,
J. Hennen, Helen
Sota˜ o No. 87–5
(PUR 89746). FRENCH GUIANA: Iracubo
canton, west of Iracubo, close to village Bellevue, side road
of road N1 to village Rocoucoua. 5u289400N, 53u169420W, 20
m, 19 Jul 2009,
L. Beenken & R. Berndt No. 19.07.09/9
(ZT
Myc 3393, PC). Saint Laurent du Maroni canton, Saint Jean,
road to Plateau des Mines, 5u249N, 54u039W, 50 m, 22 Jul
2009,
L. Beenken & R. Berndt No. 22.07.09/2
(ZT Myc
3394, PC). Mana canton, Piste Montagne de Fer, side road
of road N1 at km 200, 5u239580N, 53u339360W, 70 m, 23 Jul
2009,
L. Beenken & R. Berndt No. 23.07.09/12
(ZT Myc
3395, PC). Roura canton, route de Kaw (D6), 4u309N,
FIG. 10.
Dasyspora winteri
clade. Differences in ornament of the teliospores between the species (drawings from
type collections).
670 MYCOLOGIA
TABLE III. Measurements of teliospores and shape and size of teliospore warts in
Dasyspora
(in micrometers)
Species Host Distribution
Spore size
Projections
Lateral warts
Mean Form Height In surface view At septum
1.
D. gregaria X. cayennensis
Surinam, French
Guiana, Brazil
Amapa´
30–44 321–30 up to 10 31.5–2 hemispherical 1–2 1–2 1–2
37.1 327.2
2.
D. segregaria X. aromatica
Panama, North
Venezuela, Peru,
Trinidad
31–41 321–30 up to 10 31.5–2 hemispherical 1–2 1–2 1–2
36.8 326.5
3.
D. echinata X. aromatica
Cerrados of Brazil 35–44 324–30 up to 7 31.5–2 conical 2–3 1–2 1–2
37.9 327.1
4.
D. winteri X. sericea
Brazil, Rio de
Janeiro
30–39 320–27 up to 7 31 polygonal,
flattened
0.5–1 1–1.5 2–3 31.5–2 (3)
34.4 323.8
5.
D. frutescentis X. frutescens
var.
frutescens
Venezuela, Guyana,
French Guiana,
Brazil
28–38 318–25 up to 8 31.5 polygonal,
flattened
0.5 0.5–1 30.5–1.5 2–3 31–2
32.9 322.2
6.
D. ferrugineae X. frutescens
var.
ferruginea
French Guiana 30–36 321–24 up to 8 31.5 polygonal,
flattened
1 0.5–1.5 30.5–2 2–4 31–2
33.1 323.3
7.
D. mesoamericana X. frutescens
var.
frutescens
Mexico, Belize,
Guatemala,
Panama
29–40 321–28 up to 8 31.5 polygonal,
flattened
0.5–1 0.5–1 30.5–2 2–3 31–2
34.5 324.4
8.
D. emarginatae X. emarginata
Cerrados of Brazil,
Distrito Federal
32–38 325–27 up to 6 31.5 polygonal,
flattened
0.5(1) 1–1.5(2) 1–2 (2.5) 31–1.5
34.6 325.5
9.
D. nitidae X. nitida
French Guiana 28–40 319.5–27 up to 8 31.5 polygonal,
flattened
0.5(1) (1) 1.5–2 1.5–2 31.5–2
33.8 321.7
10.
D. guianensis X. benthamii
French Guiana 29–32 321.5–24 up to 5 32 polygonal,
flattened
0.5 1 (1.5) 1.5–3 31–1.5
30.5 322.4
11.
D. amazonica X
. cf.
amazonica
Brazil, Amazonas 25–30 318–21 up to 5 31.5 polygonal,
flattened
0.5 1–1.5 1.5–2.5 (3) 31–2
27.6 319.5
BEENKEN ET AL:
D
ASYSPORA
671
52u109W, 100 m, 27 Jul 2009,
L. Beenken & R. Berndt
No. 27.07.09/1
(ZT Myc 3396, PC). Roura canton, road N2
toRegina,Coralie,sideroadtoCacao,4u309360N,
52u239530W, 100 m, 08 Aug 2009,
L. Beenken & R. Berndt
No. 08.08.09/3
(ZT Myc 3397, PC).
Note.
Kunze established
Puccinia gregaria
on mate-
rial collected by Weigelt in Surinam and issued it in
‘‘Weigelt Exsiccati Surinam 1827’’ with a Latin
diagnosis on the label (Stevenson 1971: 503–507).
Berkeley and Curtis (1853) described
Dasyspora
as a
new genus with the single species
D. foveolata
based on
a part of Weigelt’s collection kept in the Schweinitz
herbarium and labeled
Aecidium foveolatum
by Schwei-
nitz (Stevenson 1971: 503–507, Hennen et al. 2005:
68). Therefore
D. foveolata
is a homotypic synonym of
D. gregaria
(Hennings 1896).
2. Dasyspora segregaria Beenken sp. nov. FIG.8b
MycoBank MB519382
Species characteribus generis; differt a
Dasyspora gregaria
Xylopia aromatica
hospite et sequentiis acidi nucleici ITS
LSUque.
Etymology.
A compositum from Latin
se
-(5apart,
out of) and the epithet
gregaria
to indicate segregation
of the species from
D. gregaria
(
segregare
5to
segregate, to separate).
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial on leaves,
subepidermal, erumpent, dark brown, densely aggre-
gated on bleached, yellowish leaf spots of 2–10 mm
diam, arranged in concentric circles or scattered.
Paraphyses peripheral, rarely between teliospores,
cylindrical to slightly clavate, 20–50 34–6 mm; wall
0.3–2 mm thick. Teliospores two-celled by transverse
septum, ellipsoid, constricted at the septum, 31–41 3
21–30 mm (mean 36.8 326.5 mm), cells subglobose,
one germ pore apical in the distal cell and one next to
the hilum in the proximal cell; spore wall brown, two-
layered, (1)2–3(4) mm thick, verrucose; apically and
basally with rod-like projections branched at the tip,
up to 10 31.5–2 mm; lateral warts hemispherical, 1–
2mm diam and 1–2 mm high; pedicel colorless, fragile,
breaking off close to the hilum.
Host and distribution.
On
Xylopia aromatica
(Lam.)
Mart. (5
X. grandiflora
A. St.-Hil., 5
X. cubensis
A.
Rich.); Panama, Peru, Trinidad, Venezuela (FIG.3a).
HOLOTYPE: TRINIDAD AND TOBAGO. TRINI-
DAD ISLAND: Imperial College of Tropical Agricul-
ture, Fungi of Trinidad (B.W.I.), No. 1067, Mount St
Benedict, on
Xylopia aromatica,
1 Feb 1947, R.E.D.
Baker (BPI 853915).
Additional specimens examined:
PANAMA. Fungi of
Panama collected by
F. L. Stevens No. 1102
, Bella Vista, 7
Oct 1924, (PUR 42391, BPI US0116390). PROV. CHIR-
IQUI
´: Distr. de Dolega, Los Algarrobos, road to the
woman’s prison, ca. 140 m, 18 Jun 2006,
T. Trampe
No. TT158
(ZT Myc 1302). Los Algarrobos, 21 Mar 2006,
M. Piepenbring, T. Hofmann, T. Trampe No. ppMP 722
(M
0141242). Los Algarrobos, corr. Dolega, path to rı´o
Majagua, ca. 150 m, 5 Dec 2009,
M. Piepenbring, K. Arau´z,
L. Saldan˜ a No. MP4941
(in PMA). —, 6 Mar 2007,
O.P.
Perdomo, M. Piepenbring, M. Vega, T. Hofmann No. OPP
1861
(M 0141248). PERU. SAN MARTI
´N: E. Ule, Herbar-
ium Braziliense
No. 3255
, Tarapoto, Feb 1903, (PUR 65597
5F8697). Tarapoto, Oct 1902,
E. Ule No. 3262
(B
700021602). TRINIDAD AND TOBAGO, TRINIDAD IS-
LAND: Imperial College of Tropical Agriculture, Fungi of
Trinidad (B.W.I.),
No. 721
, Erin, 16 Sep 1945,
W.T. Dale
(BPI 853916). Flora of Trinidad,
No. 420
, Buenos Ayres, in
savannah, 16 Sep 1945,
R.E.D. Baker
(S F102350 ex Herb.
Harold Owen). VENEZUELA: Mycological Explorations of
Venezuela, University of Puerto Rico, Cornell University,
Ministerio de Salubridad, Agricultura y Cria
No. 3240
, Rd.
Maracay – Guigue, Est. Aragua, 31 Mar 1939,
Chardon,
Whetzel, Mu¨ ller
(BPI 853914, PUR 77885 5F104985).
3. Dasyspora echinata Beenken & R. Berndt sp. nov.
FIGS.7,8c
5
Dasyspora foveolata
Berk. & M.A. Curtis ss.
Hennings (1895) sub
Puccinia (Dasyspora) foveolata
Berk. & M.A. Curtis
MycoBank MB519383
Species characteribus generis; differt a
Dasyspora
spp.
teliosporis lateraliter echinatis et
Xylopia aromatica
hospite.
Etymology.
After the echinate spores.
Spermogonia subepidermal, Group VI/type 5. Aecia
and uredinia unknown. Telia abaxial, rarely adaxial on
leaves, subepidermal, erumpent, dark brown, densely
aggregated on bleached, yellowish leaf spots of 1–
10 mm diam, arranged in concentric circles or
scattered. Paraphyses peripheral, rarely between telio-
spores, cylindrical to slightly clavate, 20–50 34–6(10)
mm; wall 0.3–2 mm thick. Teliospores two-celled by
transverse septum, ellipsoid, slightly constricted at the
septum, 35–44 324–30 mm (mean 37.9 327.1 mm),
cells subglobose, one germ pore apical in the distal cell
and one next to the hilum in the proximal cell; spore
wall brown, two-layered, (1)2–3(4) mm thick, echinate;
apically and basally with rod-like projections branched
at the tip, up to 7 31.5–2 mm; lateral warts conical 1–
2mm diam and 2–3 mm high; pedicel colorless, fragile,
breaking off close to the hilum.
Host and distribution.
On
Xylopia aromatica
(Lam.)
Mart.; in cerrado vegetation of Brazil (Goia´s, Mato
Grosso, Minas Gerais, Sa˜o Paulo, Tocantins) (FIG. 3a).
HOLOTYPE: BRAZIL. STATE OF TOCANTINS:
Pedra Branca (Krahoˆ Indian village), 8u189S47u389W,
ca. 250 m, on
X. aromatica
, 11 Dec 2003,
F. Freire
(HeRB 8486 in ZT Myc); ISOTYPE in herbarium F.
Freire, Brazil.
Additional specimens examined:
BRAZIL. GOIA
´S: ca. 1 km
E Aragarc¸ as, 14 Sep 1967,
G. Argent, J. Ramos, P.W.
672 MYCOLOGIA
Richards, R. Souza No. 6383
, ex Kew (PUR 85129 5
F18229). Meiaponte, Oct 1892,
Ule No. 1920
,(B70
0014165, PUR 75600 5F8700 ex Holway Collection).
MATO GROSSO: Cidade Anhandui (mun. Campo Grande)
do Cerrado, 17 Oct 1972,
G. Hatschenbach No. 30516
(M
0148197 removed from M 0148187). Coxipo´, 22 Dec 1893,
Malme
(S F102347 ex Herb. Lagerheim). ca. 6 km E
Xavantina, 52u209W, 14u449S, 25 Jul 1967,
J.A. Ratter, R.A.
de Castro No. 170
, ex Kew (PUR 85102 5F18202). MINAS
GERAIS: 34 km E Passos, 09 Nov 1976,
J.F. Hennen, M.M.
Hennen No. 76–598
(BPI 746651). SA
˜O PAULO: road
between Santa Albertina and Santa Rita do Passa quatro, 26
Oct 1897,
E. Hemmendorff No. 32
(S F102336, S F102338 ex
Herb. Lagerheim). Moji-Mirim (written as Mogy-mirim),
Mose´n
(S F102349 ex Herb. Lagerheim).
Species of the
Dasyspora winteri
clade
4. Dasyspora winteri (Pazschke) Beenken comb. nov.
FIGS. 7, 9a, 10
Basionym:
Puccinia winteri
Pazschke, Hedwigia 29:158–
159 (1890)
;
Dicaeoma winteri
(Pazschke) Kuntze, Revis. gen. pl.
(Leipzig) 3:471 (1898)
5
Dasyspora gregaria
(Kunze) Henn. p.p. ss. auct., p.p. ss.
Hennings (1896)
MycoBank MB519384
Spermogonia subepidermal, Group VI/type 5. Aecia and
uredinia unknown. Telia abaxial on leaves, subepidermal,
erumpent, dark brown, densely aggregated on bleached,
yellowish leaf spots of 2–5 mm diam, arranged in more or less
concentric circles. Paraphyses peripheral, rarely between
teliospores, cylindrical to slightly clavate, 20–40 34–7 mm;
wall often apically thickened, 0.3–2 mm thick. Teliospores two-
celled bytransverse septum, cylindrical to ellipsoid, not or very
slightly constricted at the septum, 30–39 320–27 mm(mean
34.4 323.8 mm), cells subglobose; one germ pore apical in the
distal cell and one next to the hilum in the proximal cell;
spore wall brown, two-layered, (1)2–3 mm thick, verrucose;
apically and basally with rod-like projections branched at the
tip, up to 7 31mm; lateral ornament verrucose to areolate,
flattened warts subpolygonal, 1–1.5 mmdiamandupto0.5mm
high, warts next to the septum in two rows, longitudinally
elongate, 2–3 31.5–2(3) mm, up to 1(1.5) mm high; pedicel
colorless, fragile, breaking off close to the hilum.
Host and distribution.
On
Xylopia sericea
A. St. Hil.;
Brazil, Rio de Janeiro and Minas Gerais (FIG.3b).
LECTOTYPE designated here: BRAZIL. RIO DE
JANEIRO: Rabenhorst - Winter, Fungi europaei
No. 3622
,on
Xylopia sericea
(det. D.M. Johnson),
Aug 1887,
E. Ule
(S F30078 ex Herb. Dietel).
ISOTYPES examined: S F30076, S F30077, S F30078,
BPI US0116381, M 0142440, M 0142441, NY 0028433,
NY 0028434.
Additional specimens examined:
BRAZIL. Aug 1887,
(BRUX 78306,27 ex Herb. E. Bommer & M. Rousseau,
probable isotype). RIO DE JANEIRO: Morro da Nova Cinta,
Aug 1887,
E. Ule No. 98
(BPI US0116393 ex Herb. Holway,
probable isotype). Sub
D. gregaria
, Weigelt Surinam 1827,
(NY 00035277 ex Herb. Sydow, PUR F8698 small leaf piece
removed from NY 00035277, probable isotypes). MINAS
GERAIS: Jaquitiba´(5Jequitiba´), Faz. da Serra, in cerrado,
24 Nov 1957,
E.P. Heringer No. 5834
(B 10 0375848).
Note.
Ule collected
D. winteri
apparently only once
in Rio de Janeiro, Brazil. Pazschke (1890) described
Puccinia winteri
from this collection and distributed it
in
Rabenhorst - Winter, Fungi europaei
as exsiccate
No. 3622
with a lot of doublets in several herbaria. All
these doublets are isotypes of
D. winteri
now. Some
specimens of this species could belong to the type
collection, but they have doubtful (PUR F8698, NY
35277) or incomplete (BRUX 78306.27, BPI
US0116393) data on their labels. The second sample
of it was found on a herbarium sheet (in B) of
X.
sericea
collected north of Rio de Janeiro in the
cerrados of Minas Gerais.
5. Dasyspora frutescentis Beenken sp. nov.
FIGS. 2, 9b, 10
MycoBank MB519385
Species characteribus generis; differt a
Dasyspora winteri
appendicibus polaribus teliosporarum 6(8) 31.5 mm,
areolis lateralibus 0.5–1 30.5–2 mm30.5 mmet
Xylopia
frutescens
hospite, differt a
Dasyspora
spp. sequentiis acidi
nucleici ITS, LSU, SSU et CO3.
Etymology.
After the host tree
Xylopia frutescens
.
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial on leaves,
subepidermal, erumpent, dark brown, densely aggre-
gated on bleached, yellowish leaf spots of 1–3(5) mm
diam, arranged in more or less concentric circles.
Paraphyses peripheral, rarely between teliospores,
cylindrical to slightly clavat, 20–40(50) 34–7 mm;
wall often apically thickened, 0.3–2 mm thick. Telio-
spores two-celled by transverse septum, more or less
cylindrical, not or very slightly constricted at the
septum, 28–38 318–25 mm (mean 32.9 322.2 mm),
cells subglobose, one germ pore apical in the distal
cell and one next to the hilum in the proximal cell;
spore wall brown, two-layered, (1)2–3 mmthick,
ornamented; apically and basally with rod-like projec-
tions branched at the tip, up to 6(8) 31.5 mm;
laterally verrucose to areolate, flattened warts sub-
polygonal, often equatorially elongated, 0.5–1 30.5–
2mm, up to 0.5 mm high, warts next to the septum in
two rows, longitudinally elongate, 2–3 31–2 mm, up
to 1 mm high; pedicel colorless, fragile, breaking off
close to the hilum.
Hosts and distribution.
On
Xylopia frutescens
var.
frutescens
Aubl.,
X. frutescens
var.
ferruginea
R.E. Fr.,
X.
discreta
(L. f.) Sprague & Hutch.; Venezuela, Guyana,
French Guiana, Brazil (FIG.3b).
HOLOTYPE: FRENCH GUIANA: Matoury canton,
Matoury, Masif de Mirande, 4u529N, 52u219W, 50 m,
BEENKEN ET AL:
D
ASYSPORA
673
on
Xylopia frutescens
var.
frutescens
, 15 Jul 2009,
L.
Beenken & R. Berndt No. 15.07.09/2
(in PC);
ISOTYPE: ZT Myc 3400. PARATYPES from the same
location and host:
L. Beenken & R. Berndt
No. 15.07.09/1
(in PC, ZT Myc 3399),
No. 15.07.09/
3
(in PC, ZT Myc 3401),
No. 15.07.09/4
(in PC, ZT
Myc 3402).
Additional specimens examined:
On
X. frutescens var.
frutescens
: BRAZIL. BAHIA: Mar/Apr 1936,
C. Torrend
(host
misidentified as
X. ochrantha
) (PUR 088304 removed from
IBI 2272). CEARA
´: Guaramiranga City,
F. Freire
(HeRB 8530
in ZT Myc). FRENCH GUIANA: Iracubo canton, branch
from road N1 W Iracubo, close to Bellevue, side road to
Rocoucoua, 5u289400N, 53u169420W, 20 m, 19 Jul 2009,
L.
Beenken & R. Berndt No. 19.07.09/4
(in PC, ZT Myc 3403).
Ile de Cayenne, Route Cabac¸ on, secondary forest, 1 Jun
1971,
K. Kubitzki No. 71–122
(M 0148195 removed from M
0148190). GUYANA: Fungi of British Guiana, Collected
by
F.L. Stevens No. 589
, Kartabo, 23 Aug 1922 (BPI
US0116391, BPI US0116392, NY s.n. rust fungi not present).
VENEZUELA: Mycological Explorations of Venezuela,
University of Puerto Rico, Cornell University, Ministerio
de Salubridad, Agricultura y Cria
No. 3249
, Rd. Maracay –
Guigue, Est. Aragua, 31 Mar 1939,
Chardon, Whetzel, Mu¨ ller
(BPI 853917). —,
No. 3250
(BPI 853918).
On
X. frutescens
var.
ferruginea
: VENEZUELA.
BOLIVAR: Gran Sabana, Quebrada de Jaspe, 4 Jan
1994,
R. Berndt and V. Faust-Berndt No. HeRB A-141
(in ZT Myc).
On
X. discreta
: VENEZUELA. BARINAS: Ticoporo
forest reserve, 70u409W, 7u559N, 300 m, near Rio
Michay, primary forest, 16 Mar 1964,
F.J. Breteler
No. 3729
(M 0148196, removed from M 0148192 ex
MER and WAG).
6. Dasyspora ferrugineae Beenken sp. nov.
FIGS. 9c, 10
MycoBank MB519386
Species characteribus generis; differt a
Dasyspora winteri
ornamentis teliosporarum fuscis, areolis lateralibus 0.5–1.5
30.5–2, a
Xylopia frutescens
var.
ferruginea
hospite, a
Dasyspora
spp. sequentiis acidi nucleici ITS, LSU et SSU.
Etymology.
After the host variety var.
ferruginea
of
X.
frutescens
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial on leaves,
subepidermal, erumpent, dark brown, densely aggre-
gated on bleached, yellowish leaf spots of 1–3(5) mm
diam, arranged in more or less concentric circles.
Paraphyses peripheral, rarely between teliospores,
cylindrical to slightly clavate, 20–40 34–6 mm; wall
often apically thickened, 0.3–2 mm thick. Teliospores
two-celled by transverse septum, more or less cylindri-
cal, slightly constricted at the septum, 30–36 321–24 mm
(mean 33.1 323.3 mm), cells subglobose, one germ
pore apical in the distal cell and one next to the hilum
in the proximal cell; spore wall dark brown, two-layered,
(1)2–3 mm thick, ornamented; apically and basally with
rod-like projections branched at the tip, up to 7 (8) 3
1.5 mm; lateral ornament areolate, flattened warts
subpolygonal, 0.5–1.5 30.5–2 mm, up to 1 mm high,
warts next to the septum in two rows, longitudinally
elongate, 2–4 31–2 mm, up to 1.5 mm high; pedicel
colorless, fragile, breaking off close to the hilum.
Host and distribution.
On
Xylopia frutescens
var.
ferruginea
R.E. Fr.; French Guiana (FIG.3b).
HOLOTYPE: FRENCH GUIANA: Mana canton,
road to Montagne de Fer, branch from road N1 at
km 200, 5u259200N, 53u359320W, 60 m, on
Xylopia
frutescens
var.
ferruginea
, 23 Jul 2009,
L. Beenken & R.
Berndt No. 23.07.09/10
(in PC); ISOTYPES: ZT Myc
3407. PARATYPES from the same location:
L. Beenken
& R. Berndt No. 23.07.09/5
(in PC, ZT Myc 3404),
No. 23.07.09/6
(in PC, ZT Myc 3405) and
No. 23.07.09/9
(in PC, ZT Myc 3406).
7. Dasyspora mesoamericana Beenken sp. nov.
FIGS. 9d, 10
MycoBank MB519387
Species characteribus generis; differt a
Dasyspora winteri
appendicibus polaribus teliosporarum 6(8) 31.5 mm,
areolis lateralibus 0.5–1 30.5–2 mmet
Xylopia frutescens
hospite in Mesoamerica, differt a
Dasyspora
spp. sequentiis
acidi nucleici ITS, LSU, SSU et CO3.
Etymology.
After the Mesoamerican distribution.
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial on leaves,
subepidermal, erumpent, dark brown, densely aggre-
gated on bleached, yellowish leaf spots of 1–3(5) mm
diam, arranged in more or less concentric circles.
Paraphyses peripheral, rarely between teliospores,
cylindrical to slightly clavate, 20–40 34–6 mm; wall
rarely apically thickened, 0.3–2 mm thick. Teliospores
two-celled by transverse septum, more or less cylindri-
cal, not or very slightly constricted at the septum, 29–40
321–28 mm (mean 34.5 324.4 mm), cells subglobose,
one germ pore apical in the distal cell and one next to
the hilum in the proximal cell; spore wall brown, two-
layered, (1)2–3 mm thick, ornamented; apically and
basally with rod-like projections branched at the tip, up
to 6(8) 31.5 mm; laterally verrucose to areolate,
flattened warts subpolygonal, often equatorially elon-
gated, 0.5–1 30.5–2 mm, up to 0.5 mm high, warts next
to the septum in two rows, longitudinally elongate, 2–3
31–2 mm, up to 1 mm high; pedicel colorless, fragile,
breaking off close to the hilum.
Host and distribution.
On
Xylopia frutescens
var.
frutescens
;Mexico,Belize,Guatemala,Panama
(FIG.3b).
HOLOTYPE: PANAMA: Fungi of Panama collected
by
F.L. Stevens No. 243
, Fort Sherman, on
Xylopia
674 MYCOLOGIA
frutescens
(det. D.M. Johnson), 4 Sep 1924 (PUR
42390); ISOTYPES: NY s.n., BPI US0116387, BPI
US0116396.
Additional specimens examined:
On
Xylopia frutescens
:
BELIZE. EL CAYO DISTRICT: Plants of British Honduras,
Expedition of the University of Michigan and the Carnegie
Institution of Washington, San Ignatio, 24 Jul 1936,
E.B.
Mains No. 3882
(NY s.n., BPI US0116395 ex Herbarium of
the University of Michgan). Marshall, 31 Mar 1934,
E.B.
Mains No. 1197A
(NY s.n.). GUATEMALA. IZABAL: near
(lake) Izabal (written as Tabal, cf. Lowden 1970), 23 Feb
1907,
W.A. Kellerman
(BPI US0116384, NY s.n.). Los
Amates, 90 m, 15 Mar 1905,
W.A. Kellerman No. 5330
(BPI US0116380, BPI US0116386, PUR 42392). Morales, 8
Mar 1907,
W.A. Kellerman
(BPI US0116379, BPI
US0116385). Near Quirigua´, 72–150 m, Sewell Avery
Expedition of Field Museum of Natural History, Plants of
Guatemala
No. 72261
, 26–27 Apr 1939,
P.C. Standley
(PUR
49115). PETE
´N: 40 km from Santa Elena at Laguna Pacay,
13 Nov 1965,
Antonio Molina R. No. 15733
(PUR 66110 ex
US NH256697). MEXICO. CHIAPAS: 20 km NW of
Raudales, 300 m, 1 Apr 1966,
Gonzalez-Quintero No. 3572
(PUR 64451 ex ENCB). TABASCO: km 31 carro Huiman-
guillo a la Col. Agricola ‘‘El Aguila’’, 11 Jun 1965,
Cavelaz
et al.
No. E.S.2142
(PUR 64450 ex ENCB). VERACRUZ: W
of Estacio´ n Tanconchopan, 8 Sep 1944,
Gilly & Hernandez
(PUR 60025 ex PUL 32338). PANAMA: Fungi of Panama
collected by
F.L. Stevens No. 984
, France Field, 3 Oct 1924
(BPI US0116383; BPI US0116394, NY s.n.). —,
No. 1321
,
France Field, 24 Aug 1924 (PUR 42389). —,
No. 362
,
Paitilla, 8 Sept 1924 (BPI US0116388, NY s.n.). 6 Mar 1923,
C.V. Piper
(BPI US0116389).
8. Dasyspora emarginatae Beenken sp. nov.
FIGS. 9e, 10
MycoBank MB519388
Species characteribus generis; differt a
Dasyspora winteri
appendicibus polaribus teliosporarum 6 31.5 mmet
Xylopia
emarginata
hospite, differt a
Dasyspora
spp. sequentiis acidi
nucleici ITS, LSU, SSU et CO3.
Etymology.
After the host species
Xylopia emarginata
.
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial on leaves,
subepidermal, erumpent, dark brown, scattered to
aggregated, not forming leaf spots. Paraphyses pe-
ripheral, rarely between teliospores, cylindrical to
slightly clavate, 20–40 34–6 mm; wall 0.3–2 mm thick.
Teliospores two-celled by transverse septum, more or
less ellipsoid, slightly constricted at the septum, 32–38
325–27 mm (mean 34.6 325.5 mm), cells subglobose,
one germ pore apical in the distal cell and one next to
the hilum in the proximal cell; spore wall brown, two-
layered, (1) 2–3 mm thick, ornamented; apically and
basally with rod-like projections branched at the tip, up
to 6 31.5 mm; lateral ornament verrucose to areolate,
flattened warts subpolygonal, 1–1.5 (2) mm in diame-
ter, up to 0.5 mm high, warts next to the septum in two
rows, not or slightly longitudinally elongate, 1–2 (2.5)
31–1.5 mm, up to 1 mm high; pedicel colorless, fragile,
breaking off close to the hilum.
Host and distribution.
On
Xylopia emarginata
Mart.;
Brazil (Federal District) (FIG.3b).
HOLOTYPE: BRAZIL. DISTRITO FEDERAL: Par-
que National Aguas Emendadas NE of Brasilia, forest
along small stream, 4 Aug 1974, on
Xylopia emargi-
nata
,
J.F. Hennen, Y. Ono & E.P. Heringer No. 76–
256
(PUR N6196 removed from PUR 66525c).
9. Dasyspora nitidae Beenken sp. nov. FIGS. 9f, 10
MycoBank MB519389
Species characteribus generis. differt a
Dasyspora winteri
appendicibus polaribus teliosporarum 7 31.5 mm, a
Dasyspora
spp. areolis lateralibus 1–2 mm diam,
Xylopia nitida
hospite et sequentiis acidi nucleici ITS, LSU, SSU et CO3.
Etymology.
After the host species
Xylopia nitida
.
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial and rarely
adaxial on leaves, subepidermal, erumpent, dark
brown, solitary or aggregated on weakly bleached,
yellowish leaf spots of 1–5 (8) mm diam, more or less
scattered. Paraphyses peripheral, rarely between
teliospores, cylindrical to slightly clavate, 20–45 34–
7mm; wall rarely apically thickened, 0.3–1.5 mm thick.
Teliospores two-celled by transverse septum, cylindri-
cal ellipsoid, slightly constricted at the septum, 28–40
319.5–27 mm(mean33.8321.7 mm), cells
subglobose, one germ pore apical in the distal cell
and one next to the hilum in the proximal cell; spore
wall brown, two-layered, (1)2–3 mm thick, ornament-
ed; apically and basally with rod-like projections
branched at the tip, up to 7 31.5 mm; lateral
ornament verrucose to areolate, flattened warts round
to subpolygonal, 1–2 mm (mean 1.5 mm) diam, up to
0.5 mm high, warts next to the septum in two rows,
slightly longitudinally elongate, 1.5–2(2.5) 31.5–
2mm, up to 0.1(1.5) mm high; pedicel colorless,
fragile, breaking off close to the hilum.
Host and distribution.
On
Xylopia nitida
Dun.;
French Guiana (FIG.3b).
HOLOTYPE: FRENCH GUIANA: Roura canton, road
to Kaw (D6), side road to Fougasier, Montagne de Kaw,
4u309N, 52u109W, 100 m, on
Xylopia nitida
, 28 Jul 2009,
L.
Beenken & R. Berndt No. 28.07.09/6
(in PC); ISOTYPE:
ZT Myc 3412. PARATYPES from the same location:
L.
Beenken & R. Berndt No. 28.07.09/2
(in PC, ZT Myc
3410),
No. 28.07.09/5
(in PC, ZT Myc 3411).
Additional specimens on Xylopia
cf.
nitida examined:
FRENCH GUIANA: Sinnamary canton, road Saint Elie,
branch of road N1, Montagne Chapeau, 5u17940.50N,
53u03906.70W, 50 m, 24 Jul 2009,
L. Beenken & R. Berndt
No. 24.07.09/1
(in PC, ZT Myc 3408). Roura canton, road
N2 to Regina, Foret de Tiponni, 4u279250N, 52u219020W,
BEENKEN ET AL:
D
ASYSPORA
675
200 m, 08 Aug 2009,
L. Beenken & R. Berndt No. 08.08.09/
1
(in PC, ZT Myc 3409).
Note.
The determination of the host plants from
Montagne Chapeau and Foret de Tiponni is uncertain
because the leaf indumentum is brownish in contrast
to the descriptions of
X. nitida
that has white hairs
(Fries 1931, Steyermark et al. 1995, Maas & Maas-van
de Kamer 2002). The type collections from Montagne
de Kaw fit very well to
X. nitida
.
10. Dasyspora guianensis Beenken sp. nov.
FIGS. 9g, 10
MycoBank MB519390
Species characteribus generis; differt a
Dasyspora
spp.
appendicibus polaribus teliosporarum 5 32mm,
Xylopia
benthamii
hospite et sequentiis acidi nucleici ITS, LSU, SSU
et CO3.
Etymology.
After French Guiana.
Spermogonia
subepidermal, Group VI/type 5.
Aecia
and
uredinia
unknown.
Telia
abaxial on leaves,
subepidermal, erumpent, dark brown, densely aggre-
gated on bleached, yellowish leaf spots of 3–7 mm
diameter, arranged in more or less concentric circles.
Paraphyses
peripheral, rarely between teliospores,
cylindrical to slightly clavate, 20–35 34–5 mm; wall
0.3–1.5 mm thick.
Teliospores
two-celled by transverse
septum, cylindrical to ellipsoid, not or very slightly
constricted at the septum, 29–32 321.5–24 mm (mean
30.5 322.4 mm), cells subglobose, one germ pore
apical in the distal cell and one next to the hilum in the
proximal cell; spore wall brown, two-layered, (1) 2–
3mm thick, ornamented; apically and basally with rod-
like projections branched at the tip, up to 5 32mm;
lateral ornament verrucose to areolate, flattened warts
subpolygonal, 1–1.5 mm diam, up to 0.5 mm high, warts
next to the septum in two rows, longitudinally
elongate, 1.5–3 31–1.5 mm, up to 1 mm high; pedicel
colorless, fragile, breaking off close to the hilum.
Host and distribution.
On
Xylopia benthamii
R.E. Fr.;
French Guiana (FIG.3b).
HOLOTYPE: FRENCH GUIANA: Kourou canton,
De´grad Saramaka, near waterworks, forest at river
Kourou, 5u009540N, 52u419560W, 10 m, on
Xylopia
benthamii
, 18 Jul 2009,
L. Beenken & R. Berndt
No. 18.07.09/4
(in PC); ISOTYPE: ZT Myc 3413.
11. Dasyspora amazonica Beenken sp. nov.
FIGS. 9h, 10
MycoBank MB519391
Species characteribus generis differt a
Dasyspora
spp.
teliosporis minoribus, 25–30 318–21 mm,
Xylopia
cf.
amazonica
hospite et sequentiis acidi nucleici ITS, LSU, SSU et CO3.
Etymology.
After the Amazonian region.
Spermogonia subepidermal, Group VI/type 5.
Aecia and uredinia unknown. Telia abaxial on leaves,
subepidermal, erumpent, brown, aggregated on
bleached leaf spots of 3–5 mm diam, arranged in
more or less concentric circles. Teliospores two-celled
by transverse septum, cylindrical to ellipsoid, not or
very slightly constricted at the septum, 25–30 318–
21 mm (mean 27.6 319.5 mm), cells subglobose, one
germ pore apical in the distal cell and one next to the
hilum in the proximal cell; spore wall pale brown,
two-layered, 1–2 mm thick, ornamented; apically and
basally with rod-like projections branched at the tip,
up to 5 31–1.5 mm; lateral ornament verrucose to
areolate, flattened warts subpolygonal, 1–1.5 mm
diam, up to 0.3(0.5) mm high, warts next to the
septum in two rows, longitudinally elongate,1.5–
2.5(3) 31–2 mm, up to 0.5(1) mm high; pedicel
colorless, fragile, breaking off close to the hilum.
Host and distribution.
On
Xylopia
cf.
amazonica
R.E.
Fries; Brazil (FIG.3b).
HOLOTYPE: BRAZIL. AMAZONAS: Paraiso, Ma-
deira River, on
Xylopia
cf.
amazonica
, 03 Sep 1923,
J.R. Weir
(BPI US0116382); ISOTYPE: B 700014100.
Note.
The host determination is not entirely certain
because the collection consists of young twigs with leaves
only.
KEY TO THE SPECIES OF
D
ASYSPORA
1 Lateral teliospore wall tuberculate to echinate with
hemispheric or conical warts higher than 1.5 mm,
on
Xylopia aromatica
or
X. cayennensis
............
....................
D. gregaria
clade 2
19Lateral wall ornament areolate, formed by flat-
tened, more or less polygonal warts up to 1 mm
high, warts close to the septum arranged in two
rows, slightly to distinctly longitudinally elongate
and bigger than the lateral ones, on other
Xylopia
spp. .....................
D. winteri
clade 4
2 Lateral warts conical, on
X. aromatica
.........
.................... 3.
D. echinata
29Lateral warts hemispherical ............. 3
3On
X. cayennensis
............... 1.
D. gregaria
39On
X. aromatica
.............. 2.
D. segregaria
4 Sori not on leaf spots, on
X. emarginata
......
.................. 8.
D. emarginatae
49Sori on yellow leaf spots ............... 5
5 Polar projections up to 1 mmthick,on
X.
sericea
........................ 4.
D. winteri
59Polar projections thicker, up to 2 mm ......... 6
6 Polar projections up to 5 mm long, teliospores
up to 32 324 mm .................... 7
69Polar projections up to 8 mm long, up to 1.5 mm
thick, teliospores larger ................ 8
7 Polar projections up to 2 mm thick, teliospores 29–32
321.5–24 mm, on
X. benthamii
.... 10.
D. guianensis
79Polar projections up to 1.5 mm thick, teliospores 25–30
318–21 mm, on
X. amazonica
..... 11.
D. amazonica
8 Lateral warts isodiametric, roundish to slightly
polygonal, up to 2 mm diam, ornament close to
676 MYCOLOGIA
the septum similar to the lateral one, on
X.
nitida
..................... 9.
D. nitidae
89Lateral warts distinctly polygonal, transversely
elongate, up to 1 31.5 mm, warts close to the
septum much bigger than the lateral ones, on
X. frutescens
........................ 9
9 Teliospores dark brown, lateral ornament dense, up to
1mmhigh,on
X. frutescens
var.
ferruginea
............
........................ 6.
D. ferrugineae
99Teliospores light brown, lateral ornament up to
0.5 mm high, on
X. frutescens
var.
frutescens,
rarely
on
X. frutescens
var.
ferruginea
.............. 10
10 Teliospores up to 40 328 mm, distributed in
Central America ....... 7.
D. mesoamericana
109Teliospores up to 38 325 mm, distributed in
South America ............ 5.
D. frutescentis
DISCUSSION
The genus
Dasyspora
has been regarded as monotypic
since its erection in 1853. In the present study we
investigated numerous specimens of the genus from
its entire geographic range and host spectrum. We
were able to show that
Dasyspora
is not monotypic but
comprises 11 species, which fall into two major clades.
Species delimitation within Dasyspora.—
We com-
bined morphological characters, a multiple gene
phylogeny and ecological niches (host plants and
habitats) to recognize species in
Dasyspora
. With this
approach we follow the species concept of Harrington
and Rizzo (1999), who define a species as ‘‘the
smallest aggregation of populations with a common
lineage that share unique, diagnosable phenotypic
characters’’. We also consider the genealogical
concordance phylogenetic species recognition
(GCPSR) criteria of Taylor et al. (2000) by comparing
phylogenetic analyses of the single DNA regions. We
did not find any conflicts (i.e. differences in tree
topology with high bootstrap support) among the
single-gene trees. In principle our results are not
contrary to the GCPSR criteria. However the present-
ed species delimitation was not supported in all cases
by the molecular data that were not always congruent
with the morphological observations. We found some
samples to be indistinguishable by morphology but
differing in their DNA sequences (
D. gregaria
vs.
D.
segregaria
and
D. frutescentis
vs.
D. mesoamericana
).
Hence it is apparent that the genus
Dasyspora
includes cryptic species. In contrast some collections
had identical or very similar sequences but differed
conspicuously in their teliospore ornamentation and
could be identifed reliably by microscopic examina-
tion (
D. gregaria
vs.
D. echinata
and
D. frutescentis
vs.
D. ferrugineae
). Two species complexes comprising
very closely related species were identified within
Dasyspora
(FIG. 5): (i)
Dasyspora gregaria
agg. (equiv-
alent to
D. gregaria
clade) including
D. gregaria
,
D.
segregaria
and
D. echinata
; (ii)
D. frutescentis
agg.
(part of the
D. winteri
clade) including
D. frutescentis
and
D. ferrugineae
.
We decided to describe all taxa in these complexes at
species rank and not at a subordinate rank because we
consider spore morphology an important character to
delimit species in rust fungi. This is especially legitimate
because morphologies are strongly correlated to host
preferences. However differences between sequences of
the species of each complex are low or lacking. Similar
or identical ITS sequences among closely related species
also have been observed in several other fungal genera
even when the species delimitation is well supported by
morphological, ecological or other molecular data
(Bruns 2001, Burnett 2003, Gru¨nig 2004, Harrington
and Rizzo 1999, Harrington et al. 2002, Maijala et al.
2003, O’Donnell et al. 1998, Samuels et al. 2010, Vialle
et al. 2009). Other studies (Scholler et al. 2011, Liang
et al. 2006) argue that morphologically identical rust
species, which infect different host species but share
identical ITS and LSU rDNA, should not be regarded as
different but instead as a single species. In contrast
D.
echinata
and
D. gregaria
,aswellas
D. frutescentis
and
D.
ferrugineae
, are not morphologically identical species.
The Dasyspora gregaria clade.—
We found
D. gre-
garia
on its type host
X. cayennensis
in French Guiana.
The host identity and the geographical proximity to
the type locality in Surinam strongly suggest that our
material actually represents the rust species on which
Kunze based
D. gregaria
. Because no DNA could be
extracted from type material of
D. gregaria
we
consider DNA from the French Guiana specimen as
authentic for the species. The conspicuous morpho-
logical differences in teliospore ornament between
D.
gregaria
and
D. echinata
justify the description of a
new species. However they cannot be separated by the
investigated sequences. To avoid
D. gregaria
becom-
ing paraphyletic to
D. echinata
(FIGS.4–6) it is
necessary to segregate
D. segregaria
as a distinct
species. Their hosts and geographic distributions
furthermore distinguish the three species. The
observed differences between the induments of the
hosts of
D. echinata
and
D. segregaria
might indicate
that
X. aromatica
comprises two distinct taxa; one
restricted to the cerrados of Brazil has erect bent leaf
hairs, the second occurs in northern South America
and Central America and possesses appressed straight
hairs. Fries (1931) wrote in his revision of Annona-
ceae that
X. aromatica
was variable regarding its
hairiness. He noted that the forms with appressed
hairsweremorecommoninthenorthofits
distribution area.
BEENKEN ET AL:
D
ASYSPORA
677
The Dasyspora winteri clade.—
Eight species were
distinguished based on sequence differences. Mor-
phological characters and host preferences support
this with one exception.
Dasyspora mesoamericana
and
D. frutescentis
belong to different lineages, but they
are morphologically indistinguishable. We consider
them to be cryptic species. Roy et al. (1998) identified
cryptic species in the
Puccinia monoica
complex and
Benett et al. (2011) in
Melampsora
but without
naming them. To our knowledge
D. mesoamericana
and
D. frutescentis
are the first taxonomically de-
scribed and named cryptic species in Puccinales. The
observed morphological similarity of these two
distantly related
Dasyspora
species is remarkable. In
contrast
D. ferruginea
, which is not well separated
from
D. frutescentis
by the investigated sequences,
shows a spore ornament well distinguishable from its
sister species. In the group of specimens we assigned
to
D. nitidae
the observed sequence differences might
indicate the presence of two distinct
Dasyspora
species. The differences in the indumentum of the
host leaves suggest two different host taxa and would
support a split of
D. nitidae
as well. Similar conditions
were found for
D. frutescentis
and
D. ferrugineae
on
two varieties of
X. frutescens
and were hypothesized
for
D. echinata
and
D. segregaria
on
X. aromatica
.In
contrast the observed uniform morphology of
D.
nitidae
led us to assign all specimens to a single
species currently.
D. nitidae
likely will be split into two
morphologically similar sister species when more
samples are available and the relations of
X
. cf.
nitida
are clarified.
Host and geographic distribution.—
In contrast to its
pantropical distributed host genus
Xylopia
,
Dasyspora
is restricted to the tropical Americas (FIG. 3). The
center of diversity with five species was found in the
Guianas. One is known from the middle of the
Amazonian rainforest. Three species occur south of
the Amazonian Basin in the cerrados and bordering
Atlantic Forest. Only one member of each main clade
is distributed in Central America. One may assume
that each rust species has a distribution range similar
to its host plant, but most
Dasyspora
species are
known only from the type locality and sometimes a
few additional sites. To date only the three species
of the
D. gregaria
clade,
D. mesoamericana
and
D.
frutescentis
have been sampled well enough to
estimate their distribution and host range. All
collections of these species originated from the same
Xylopia
taxon and the same geographic region
(FIGS. 3, 5). Therefore a host specifity and a distinct
distribution area could be hypothesized at least for
these
Dasyspora
species.
Cospeciation and host jump in Dasyspora.—
In that
Dasyspora
is strictly associated with American mem-
bers of
Xylopia
coevolution between parasite and host
could be hypothesized. A comparison of the phylog-
eny of
Dasyspora
with the known systematics of
Xylopia
(Fries 1931, Maas et. al 1993) indicates cases of
cospeciation and of host jumps too (cf. Roy 2001).
The sister species
D. frutescentis
and
D. ferrugineae
occur on two varieties of
X. frutescens
. Therefore their
speciation might have paralleled the split of their host
taxa. It is interesting to note here that Fries (1931)
suggested that
X. frutescens
var.
ferruginea
might be
elevated to species rank. Furthermore an ancient host
jump could explain that
D. mesoamericana
and
D.
frutescentis
, which are not closely related, had
colonized
X. frutescens
var.
frutescens
independently,
once in Central America and once in northern South
America. Ritz et al. (2005) demonstrated a similar
host jump within the genus
Phragmidium,
in which
two species of different molecular linages occur on
the same
Rosa
species.
Cospeciation could explain why the species of the
Dasyspora gregaria
clade exclusively parasitize
X.
aromatica
and the close related
X. cayennensis
.
Xylopia
aromatica
from the Brazilian cerrados differs in some
characters from the plants in the humid climates in
northern South America. It shows adaptions of leaves
to the severe dry season in their habitat expressing a
dense hair cover (Bieras and Sajo 2009). Likewise it
could be speculated that the spiny teliospore orna-
mentation of
D. echinata
is a co-adaption to the drier
habitat in the cerrados (cf. Savile 1971) or to other
unknown environmental factors.
X. nitida
belongs to
the
X. aromatica
complex but its rust
D. nitidae
belongs to the
D. winteri
clade, according to Fries
(1931). Perhaps this is a further example of a host
jump in
Dasyspora
. A final conclusion of evolutionary
trends in
Dasyspora
will be possible when a molecular
phylogeny of
Xylopia
is available.
Position of Dasyspora within Pucciniales.—
Hitherto
Dasyspora
has been placed in the family Uropyxida-
ceae because of the two-celled teliospores and
spermogonial type (Cummins and Hiratsuka 1983,
2003). In our results
Dasyspora
did not show a close
relationship to species of
Porotenus
and
Tranzschelia
of Uropyxidaceae, indicating that the family as
defined by Cummins and Hiratsuka (2003) is poly-
phyletic. This is in accordance with results obtained
by Aime (2006). Unfortunately
Uropyxis
J. Schro¨t., the
type genus of Uropyxidaceae and its type species
U.
amorphae
(Curtis) J. Schro¨t., have not been included
in any molecular analysis of rust fungi to date and the
status and circumscription of the family remain
uncertain.
Puccinia popowiae
and
Diorchidium poly-
678 MYCOLOGIA
althiae,
which appeared as sister group of
Dasyspora
,
likewise occur on Annonaceae,
P. popowiae
on
Monanthotaxis caffra
Verdc. (5
Popowia caffra
Benth.) in South Africa and
D. polyalthiae
on
Polyalthia longifolia
(Sonn.) Thwaites in India. Like
Dasyspora
, both species have two-celled and orna-
mented teliospores—
P. popowiae
with a horizontal,
often oblique septum and
D. polyalthiae
with vertical,
rarely oblique septum. They are autoecious as well but
form aecia and lack in spermogonia in contrast to the
microcyclic
Dasyspora
(Sydow and Sydow 1915, Petch
1912 [sub
Aecidium polyalthiae
Petch], Cooke 1882,
Doidge 1932, pers obs). The molecular data indicate
that
P. popowiae
does not belong to the genus
Puccinia
Pers. or other Pucciniaceae. According to
Cummins and Hiratsuka (2003),
Diorchidium
Kalchbr. belongs to Raveneliaceae, but
D. polyalthiae
does not group together with
Ravenelia
in our
phylogram. Due to these discrepancies the systematic
positions of
Dasyspora
,
Puccinia popowiae
and
Diorch-
idium polyalthiae
within rust fungi could not be
settled with available data.
ACKNOWLEDGMENTS
The authors thank the curators of B, BPI, BRUX, M, NY,
PMA, PUR and S for loan of specimens. M. Piepenbring
(Frankfurt) is thanked for specimens from Panama. D.M.
Johnson (Delaware) helped kindly to identify host plants. We
are much obliged to the curators of CAY, especially M.-F.
Prevost, for their hospitality and help in French Guiana. The
first author thanks T. Torossi and A.M. Minder for their
technical assistance during his molecular work in the lab of
the Genetic Diversity Centre ETH Zurich. L. Lannes (Zurich)
gave us useful information about the cerrados of Brazil. We
thank M. Oberhofer (Zurich) for her helpful comments to
the manuscript. The Swiss National Fund (SNF) financially
supported this study (Project 31003A-116095).
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