Mycol Progress (2011) 10:175–187
DOI 10.1007/s11557-010-0687-0
ORIGINAL ARTICLE
Taxonomy and phylogeny of Puccinia lagenophorae: a study
using rDNA sequence data, morphological and host range
features
Markus Scholler & Matthias Lutz & Alan R. Wood &
Gregor Hagedorn & Mechthilde Mennicken
Received: 15 January 2010 / Revised: 28 May 2010 / Accepted: 16 June 2010 / Published online: 6 July 2010
# German Mycological Society and Springer 2010
Abstract Puccinia lagenophorae is a rust fungus originating from Australasia which has spread throughout the
world. A phylogenetic analysis of taxa related to this
species was performed using rDNA (LSU, ITS) sequence
data. The analyses revealed a well-supported cluster
Electronic supplementary material The online version of this article
(doi:10.1007/s11557-010-0687-0) contains supplementary material,
which is available to authorized users.
M. Scholler (*)
Staatliches Museum f. Naturkunde, Abt. Biowissenschaften,
Erbprinzenstr. 13,
76133 Karlsruhe, Germany
e-mail: scholler@naturkundeka-bw.de
M. Lutz
Lehrstuhl f. Organismische Botanik,
Institut f. Evolution und Ökologie, Universität Tübingen,
Auf der Morgenstelle 1,
72076 Tübingen, Germany
A. R. Wood
ARC-Plant Protection Research Institute,
P. Bag X5017,
Stellenbosch 7599, South Africa
G. Hagedorn
Julius Kühn-Institute (JKI),
Federal Research Centre for Cultivated Plants,
Institute for Epidemiology and Pathogen Diagnostics,
Königin-Luise-Str. 19,
14195 Berlin, Germany
M. Mennicken
Lehrstuhl Organismische Botanik,
Institut für Evolution und Ökologie, Universität Tübingen,
Auf der Morgenstelle 1,
72076 Tübingen, Germany
including all specimens of P. lagenophorae. By evaluating
morphological and sequence data, the species is taxonomically re-defined and a list of synonyms is provided.
Puccinia distincta on Bellis perennis, a species recently
separated from P. lagenophorae, P. saccardoi, a species on
the Goodeniaceae, and P. byliana, a species so far only
known from South Africa, are reduced to synonymy in P.
lagenophorae, as are several other species. Our analysis
indicates that P. lagenophorae is likely not derived from the
northern hemisphere species P. obscura, but from a species
from Australia host-alternating between Asteraceae (aecial
host) and Cyperaceae/Juncaceae (telial host). Another
related species, P. stylidii (on Stylidium sp., Stylidiaceae)
may have been derived from the same parental species as P.
lagenophorae. From ontogenetical and morphological
studies, the presence of pycnia could not be confirmed in
the life cycle of this species, and the width of the pedicel of
teliospores at the point of attachment was found to be
highly variable and not a taxonomic character. The number
of known host species is approximately 150, including 41
new host plants recorded herein.
Keywords Molecular phylogeny . Morphology . Puccinia
lagenophorae and Puccinia distincta and Puccinia
saccardoi . Neomycete
Introduction
Puccinia lagenophorae Cooke (Pucciniales; Pucciniaceae)
is an autoecious rust fungus which is native to Australasia
(Wilson et al. 1965). It is one of a complex of species
that have radiated in Australia on the Asterales (van der
Merwe et al. 2008). Today, this neomycete species has an
176
almost worldwide distribution (Europe, South America,
North America, North Africa, Southwest Asia). So far,
about 60 host species are known (Farr and Rossman
2009), all belonging to the subfamily Asterioideae (family
Asteraceae). Puccinia lagenophorae is characterized by a
demicyclic life cycle with repeating aeciospores which
have hyaline pore plugs (type 5 aeciospores of Savile
1973) and telia with smooth apically thickened telio- and
mesospores and no paraphyses. McAlpine (1906) considered all taxa with more or less identical morphology and
life cycle but occurring on different host plant genera
in Australia as different species. Based on biometrical
studies and inoculation experiments, Viennot-Bourgin
(1964) and Wilson et al. (1965) indicated that the
species can neither be split into “morphological” nor into
“biological” species. Consequently, Wilson et al. (1965)
combined them under the oldest name, Puccinia lagenophorae (type host species: Lagenophora billardierei Cass.,
tribe Asterae) within which were included P. allanii G.
Cunn., P. calendulae McAlpine, P. calocephali McAlpine,
P. cinerariae McAlpine, P. distincta McAlpine, P. erechtitis
McAlpine, P. hypochoeridis McAlpine (non P. hypochoeridis
Oudem.), P. macalpinei P. Syd. & Syd., P. tasmanica Dietel,
and P. terrieriana Mayor (Cunningham 1931; Wilson et al.
1965). Wilson et al. (1965) also indicated that other rust
species on other genera of the Asteraceae from Australasia
with identical life cycle and spore morphology, such as P.
brachycomes McAlpine, would likely prove to also belong
to this species. Later, this species concept was confirmed by
other authors such as Walker (1983) and Scholler (1993,
1997) after detailed morphological studies. Walker (1983)
also indicated that P. byliana Dippenaar, a species from
South Africa, may be related or even conspecific with P.
lagenophorae.
In several publications, Weber and coauthors (Weber and
Tilston 1999; Weber et al. 1998, 2003; Preece et al. 2000)
differentiated the rust on Bellis perennis L. in Europe from
P. lagenophorae (on Senecio vulgaris L.) and re-established
P. distincta (Weber et al. 1998), a species described from
Australia on B. perennis. According to the authors, P.
distincta, in contrast to P. lagenophorae on Senecio
vulgaris, is not hosted by S. vulgaris, does not form pycnia,
three-celled teliospores, or average pedicel widths above
7 μm. Later, Weber et al. (2003) confirmed their findings
by presenting differences between the Bellis and the
Senecio rust in three positions of the ITS sequence. This
differentiation, however, was not generally accepted (e.g.,
Koike and Scholler 2001; Bruckart 2003; Henricot and
Denton 2005).
A phylogenetic analysis using rDNA sequence data (LSU,
ITS1-5.8 S-ITS2) and comparative morphological studies of
P. lagenophorae and related species were performed to
resolve these open questions and conflicting views.
Mycol Progress (2011) 10:175–187
Materials and methods
Morphology
Spores from dried herbarium specimens were mounted in
50% lactic acid aqueous solution, gently heated, and
examined using a Nikon Eclipse E600 (specimens from
South African) or a Zeiss Axioskop 2 Plus Microscope (all
other specimens) at ×1,000 magnification. Generally,
measurements of 25–100 spores of each spore stage were
made of selected specimens. For determining the percentage of three-celled versus two-celled teliospores, 100 spores
were evaluated per specimen. The width of the pedicel of
teliospores at the point of attachment was measured for
10 teliospores from each of five telia on the holotype of
P. lagenophorae and from other specimens on miscellanous
host plants. The data were rounded to the nearest 0.5 μm. A
multiple range test (Ryan-Einot-Gabriel-Welsch, alpha=
0.05) was performed on these data. The presence of
loosely adherent chains of aeciospores with hyaline
(pore) plugs was taken as diagnostic of this species
when no telia were observed on the specimens (Baka
1996; Littlefield et al. 2005). For comparison with
P. lagenophorae, published descriptions were evaluated
(Cunningham 1931; Littlefield et al. 2005; McAlpine 1906;
Scholler 1993, 1997; Viennot-Bourgin 1964; Wilson et al.
1965), and material from K, KR, PDD, PUR and VPRI
were examined. Herbarium acronyms are from Holmgren
et al. (1981).
Specimens examined
Selected specimens studied for morphological characters
and host range are listed in the electronic supplementary
material, Appendix A.
Molecular analyses
Details of herbarium specimens from which genomic DNA
was isolated are listed in Table 1. For methods of isolation
and crushing of fungal material, DNA extraction, amplification, purification of PCR products, sequencing, and
processing of the raw data, see Lutz et al. (2004). For all
specimens, we determined sequences of the 5′-end of the
nuc-LSU rDNA including the domains D1/D2 (LSU).
Since only the ITS of the closest relatives of the P.
lagenophorae complex are available in GenBank, we
additionally determined base sequences of the ITS1–
5.8 S–ITS2 region of the nuc-rDNA for selected specimens
(see Table 1). The LSU was amplified using the primer pair
NL1 and NL4 (O’Donnell 1992, 1993); the ITS was
amplified using the primer pair ITS1f/ITS5 and ITS4
(Gardes and Bruns 1993; White et al. 1990). For amplifi-
Mycol Progress (2011) 10:175–187
177
Table 1 List of sequenced specimens with host plants, DNA isolation numbers, GenBank accession numbers, and reference specimens (acronyms
of public herbaria according to Index Herbariorum)
Species
Host
DNA
isolation no.
GenBank acc. no.
(ITS/LSU)
Reference specimens
Aecidium sp.
Ammobium alatum R. Br.
ml1280
FJ655859/FJ669219
Australia, New South Wales, Australian
Capital Territory, Canberra, Acton,
courtyard of the Botany and Zoology
building; 25.V.2006, leg. M. van der
Merwe; KR15599, duplicate: DAR 77116
Puccinia lagenophorae
Cooke
Amellus microglossus
DC.
ml999
-/FJ669235
Puccinia lagenophorae
Cooke
Amellus nanus DC.
ml1000
-/FJ669236
Puccinia lagenophorae
Cooke
Arctotis fastuosa Jacq.
ml1001
-/FJ669237
Puccinia lagenophorae
Cooke
Bellis perennis L.
ml867
FJ655863/FJ669223
Puccinia lagenophorae
Cooke
Bellis perennis L.
ml868
FJ655862/FJ669222
Puccinia lagenophorae
Cooke
Bellis perennis L.
ml870
FJ655861/FJ669221
Puccinia lagenophorae
Cooke
Puccinia lagenophorae
Cooke
Bellis perennis L.
ml1139
FJ655860/FJ669220
Bellis perennis L.
ml1274
FJ655871/FJ669231
South Africa, Western Cape Province,
BIOTA-observatory at Flaminkvlakte; 06.
IX.2002, leg. M. Mennicken No. RSA
114; PREM 59333
South Africa, Northern Cape Province,
Richtersveld National Park, BIOTAobservatory at Koeroegapvlakte; 15.
IX.2002, leg. M. Mennicken No. RSA
160; PREM 59350
South Africa, Western Cape Province,
BIOTA-observatory at Luiperskop 211,
07.IX.2002, leg. M. Mennicken No. RSA
124; PREM 59338
USA, California, Monterey County, Salinas;
26.III.2001, leg. S. Koike; KR 8475,
duplicate: PUR N1126
Germany, Schleswig-Holstein, LübeckIsraelsdorf; 09.VIII.1998, leg. A. Schmidt;
KR 8454, duplicate: PUR N2501
Germany, Baden-Württemberg, nördliches
Oberrhein-Tiefland, Karlsruhe, Nymphengarten; 17.VII.2003, leg. M. Scholler; KR
12156
Germany, Rheinland-Pfalz, Kaiserslautern;
19.X.1999, leg. R.W.S. Weber; K(M) 92033
New Zealand, Mid Canterbury,
Christchurch, Riccarton; 24.III.1998, leg.
A. J. Healy 98/10; PDD 70633
Puccinia lagenophorae
Cooke
Calendula officinalis L.
ml1191
-/FJ669250
Puccinia lagenophorae
Cooke
Dimorphotheca pluvialis
(L.) Moench
ml869g
FJ655864/FJ669224
Puccinia lagenophorae
Cooke
Dimorphotheca sinuata
DC.
ml866
FJ655865/FJ669225
Puccinia lagenophorae
Cooke
Felicia merxmuelleri
Grau.
ml1004
-/FJ669238
Puccinia lagenophorae
Cooke
Felicia tenella (L.) Nees.
ml1005
FJ655866/FJ669226
Germany, Mecklenburg-Vorpommern,
Greifswald; 22.VII.1992, leg. M. Scholler;
KR 8443, duplicate: PUR N1174
South Africa, Western Cape, between
Clanwilliam and Calvinia, near
Brandewyn River, Traveller’s Rest guest
farm, Agterpakhuis; 07.XI.2001, leg. A. R.
Wood 300; PREM 57975, duplicate:
KR8441, PUR N2518
South Africa, Western Cape, between
Clanwilliam and Calvinia, near
Brandewyn River, Traveller’s Rest guest
farm, Agterpakhuis; 13.VII.2001, leg. A.
R. Wood 302; PREM 57977, duplicate:
KR 8444, PUR N2516
South Africa, Northern Cape Province,
BIOTA-observatory at Quaggafontein 478,
11.IX.2002, leg. M. Mennicken No. RSA
143; PREM 59346
South Africa, Western Cape Province,
BIOTA-observatory at Riverlands, 20.
IX.2002, leg. M. Mennicken No. RSA
206; PREM 59356
178
Mycol Progress (2011) 10:175–187
Table 1 (continued)
Species
Host
DNA
isolation no.
GenBank acc. no.
(ITS/LSU)
Reference specimens
Puccinia lagenophorae
Cooke
Gazania krebsiana Less.
ml1006
FJ655867/FJ669227
Puccinia lagenophorae
Cooke
Lagenophora lanata A.
Cunn.
ml1275
-/FJ669239
Puccinia lagenophorae
Cooke
Lagenophora stipitata
(Labill.) Druce
ml1279
-/FJ669251
Puccinia lagenophorae
Cooke
Nestlera biennis (Jacq.)
Spreng.
ml1007
-/FJ669240
Puccinia lagenophorae
Cooke
Osteospermum pinnatum
(Thunb.) T. Norl.
ml1011
FJ655868/FJ669228
Puccinia lagenophorae
Cooke
Senecio arenarius
Thunb.
ml1012
FJ655869/FJ669229
Puccinia lagenophorae
Cooke
Senecio glaucus L. ssp.
coronopifolius (Maire)
Alex.
Senecio inaequidens DC.
ml1192
-/FJ669246
ml988
-/FJ669244
Puccinia lagenophorae
Cooke
Puccinia lagenophorae
Cooke
Senecio inaequidens DC.
ml1198
-/FJ669247
Senecio niveus (Thunb.)
Willd.
ml1013
-/FJ669245
Puccinia lagenophorae
Cooke
Puccinia lagenophorae
Cooke
Senecio rodriguezzi
Willk. ex Rodrig.
Senecio vulgaris L.
ml1194
-/FJ669248
ml872
FJ655870/FJ669230
Puccinia lagenophorae
Cooke
Senecio vulgaris L.
ml1277
FJ655872/FJ669232
Puccinia lagenophorae
Cooke
Tripteris amplectens
Harv.
ml1008
-/FJ669241
Puccinia lagenophorae
Cooke
Tripteris clandestina
Less.
ml1009
-/FJ669242
Puccinia lagenophorae
Cooke
Tripteris microcarpa
Harv.
ml1010
-/FJ669243
South Africa, Western Cape Province,
BIOTA-observatory at Flaminkvlakte 111,
06.IX.2002, leg. M. Mennicken No. RSA
121; PREM 59336
New Zealand, Auckland, Kawau Island,
North Cove; 18.X.2003, leg. R.E. Beever;
PDD 78352
Australia, New South Wales, Druce,
Budwang National Park; 28.XII.2006, leg.
M. van der Merwe; KR15600, duplicate:
DAR 77117
South Africa, Western Cape Province,
BIOTA-observatory at Rocherpan Nature
Reserve, 21 XI 2001, leg. M. Mennicken
No. RSA 19, PREM.
South Africa, Western Cape Province,
BIOTA-observatory at Moedverloren 208,
05.IX.2002, leg. M. Mennicken No. RSA
110; PREM 59332
South Africa, Northern Cape Province,
Richtersveld National Park, BIOTAobservatory at Numees, 03.IX.2002, leg.
M. Mennicken No. RSA 150; PREM
59349
Germany, Berlin, Berlin-Dahlem, testing
ground of the Biologische Bundesanstalt;
04.VIII.1997, leg. M. Scholler; KR 14873
Germany, Nordrhein-Westfalen, Siegburg;
09.X.1998, leg. G. Schmitz; KR 8467,
duplicate: PUR N2511
Germany, Nordrhein-Westfalen, Bonn; 19.
IX.1997, leg. M. Scholler; KR 14883
South Africa, Northern Cape Province,
BIOTA-observatory at Remhoogte 416,
08.IX.2002, leg. M. Mennicken No. RSA
130; PREM 59341
Spain, Mallorca, Cap Formentor; 28.
IV.1997, leg. A. Rubner; KR 14636
USA, Oklahoma, Wagoner County, SE
Tulsa, nursery; 03.VI.2004, leg. L.
Littlefield; KR 8500
New Zealand, Mid Canterbury, Christchurch,
Sumner, Kinsey Terrace; 11.IX.2005, leg.
E. H. C. McKenzie; PDD 83803
South Africa, Northern Cape Province,
BIOTA-observatory at Quaggafontein 478,
11.IX.2002, leg. M. Mennicken No. RSA
140; PREM 59345
South Africa, Western Cape Province,
BIOTA-observatory at Moedverloren 208,
05.IX.2002, leg. M. Mennicken No. RSA
109; PREM 59331
South Africa, Northern Cape Province,
Richtersveld National Park, BIOTAobservatory at Numees, 13.IX.2002, leg.
M. Mennicken No. RSA 149; PREM
59348
Puccinia lagenophorae
Cooke
Mycol Progress (2011) 10:175–187
179
Table 1 (continued)
Species
Host
DNA
isolation no.
GenBank acc. no.
(ITS/LSU)
Reference specimens
Puccinia lagenophorae
Cooke
Vernonia cinerea (L.)
Less.
ml1201
-/FJ669249
Puccinia obscura J.
Schröt.
Luzula campestris (L.)
DC.
ml1140
FJ655873/FJ669233
Puccinia obscura J.
Schröt.
Luzula sylvatica (Huds.)
Gaudin
ml1137
FJ655874/FJ669234
Germany, Berlin, Berlin-Dahlem, 19.
IX.1997, leg. M. Scholler; KR 14878
(artificial inoculation with inoculum from
Senecio)
England, Devon, Liddecombe, Guinspound,
Challacombe Farm; 16.IX.1999, leg. J.
Webster; K(M) 92036
Germany, Baden-Württemberg, SSW
Oppenau, Hohbruch, Ibacher Holzplatz,
18.VII.2005, leg. M. Scholler; KR 14322
cation of both regions, we adjusted the annealing temperature to 45°C. DNA sequences prepared in the course of
this study were deposited in GenBank; accession numbers
are given in Table 1.
To infer phylogenetic relationships among the analyzed
rust specimens, we also analyzed the most similar sequences available in GenBank according to a blast search
(Altschul et al. 1997): Aecidium brachycomes Petr.
EF635896 (Morin et al. 2009); Puccinia dioicae Magnus
EF635897 (Morin et al. 2009); P. lagenophorae EF212446,
EF212447 (Bruckart et al. 2007); AY808060 (Henricot and
Denton 2005); AY852264 (Littlefield et al. 2005);
EF635886, EF635889, EF635890, EF635887, EF635888,
EF635891, EF635892, EF635893, EF635894 (Morin et al.
2009); AF468041, AF468040 (Weber et al. 2003);
EU391656 (Sundelin et al. 2008); P. obscura J. Schröt.
AF468042 (Weber et al. 2003); P. rupestris Juel EF635898
(Morin et al. 2009); P. stylidii McAlpine EF635895 (Morin
et al. 2009); and P. vaginatae Juel EF635901, EF635902
(Morin et al. 2009).
Both LSU and ITS sequences were aligned with MAFFT
6.611 (Katoh et al. 2002, 2005; Katoh and Toh 2008) using
the L-INS-i option. Both alignments were used to count
base differences between the sequences. The entire ITS
alignment (length: 606 bp; variable sites: 106) was used
throughout its length for phylogenetic analyses. We avoided
both manipulation of the alignment by hand or manual
exclusion of any positions as recommended by Giribet and
Wheeler (1999) and Gatesy et al. (1993).
For phylogenetic analyses, we used both neighbourjoining analysis and a Bayesian approach. For neighbourjoining, the data were first analysed with Modeltest 3.7
(Posada and Crandall 1998) to find the most appropriate
model of DNA substitution. The hierarchical likelihood
ratio test proposed the HKY+G DNA substitution model.
Bootstrap values were calculated for 1,000 replicates. For
Bayesian analysis, we used a Markov chain Monte Carlo
(MCMC) technique as implemented in the computer
program MrBayes 3.1.2 (Huelsenbeck and Ronquist 2001;
Ronquist and Huelsenbeck 2003). Four incrementally
heated simultaneous Markov chains were run over
2,000,000 generations using the general time-reversible
model of DNA substitution with gamma-distributed substitution rates and estimation of invariant sites, random
starting trees and default starting parameters of the DNA
substitution model as recommended by Huelsenbeck and
Rannala (2004). Trees were sampled every 100th generation resulting in an overall sampling of 20,001 trees. From
these, the first 4 ,001 trees were discarded (burn-in=4,001).
The trees sampled after the process had reached stationarity
(16,000 trees) were used to compute a 50% majority rule
consensus tree to obtain estimates for the a posteriori
probabilities of groups of species. This Bayesian approach
of phylogenetic analysis was repeated four times to test
the independence of the results from topological priors
(Huelsenbeck et al. 2002). Trees were rooted with Puccinia
rupestris (EF635898).
Results
Morphology
The width at the point of attachment of the pedicel to the
distal teliospore cell was highly variable in the material
studied (Table 2). The average pedicel width of the
holotype of P. lagenophorae is slightly smaller than the
width of P. distincta on B. perennis according to Weber et
al. (1998). The average width of all specimens studied was
7.0 μm (± SE 0.05, n=750) with a minimum and maximum
of 3.5 and 12 μm, respectively. Pedicels from individual
specimens have significant width differences (Table 2). No
pycnia were observed. Three-celled teliospores occured in
variable proportions in material from Europe, Australia and
North America. None were observed in any specimen from
South Africa examined (not all specimens were examined
180
Table 2 Minimum, maximum
and mean width (in μm) of the
junction between teliospore
and pedicel for teliospores of
Puccinia lagenophorae
a
Means followed by different
letters are significantly different
(Ryan-Einot-Gabriel-Welsch
multiple range test, alpha=0.05).
Means not followed by letters
were obtained from the literature
and were not included in the
analysis
b
Holotype of Puccinia
lagenophorae
c
Sori on leaves
d
Sori on stems
e
Lectotype of Puccinia byliana
Mycol Progress (2011) 10:175–187
Host plant and specimen
Min.–max.
Meana
n
SD
Bellis perennis (KR 8456)
Lagenophora billardieri (K(M) 162111)b
Bellis perennis (KR 14476)
Bellis perennis (from Weber et al. 1998)
Lagenophora stipitata (VPRI 3803)c
Lagenophora stipitata (VPRI 3803)d
Steirodiscus tagetes (PREM 57925)
Senecio sp. (PREM 57924)
Senecio littoreus (PREM 46293)e
Emilia spp (from Henricot and Denton 2005)
Senecio vulgaris (KR 8483)d
Amellus asteroides (PREM 57925)
Bellis perennis (KR 8477)
Senecio vulgaris (KR 8483)c
Senecio vulgaris (from Weber et al. 1998)
3.5 – 8.5
4.5–10.0
4–9
5–9
4.5–8.5
5–10
5–10.5
5–9
5.5–10
5.98
6.17
6.19
6.6
6.69
7.09
7.15
7.16
7.22
7.4
7.55
7.71
7.85
8.33
8.4
100
50
100
50
50
50
50
50
50
30
50
50
100
50
50
0.97
0.97
1.1
for this character). No three-celled teliospores were observed on Amellus asteroides (L.) Druce., Arctotheca
populifolia (Bergius) Norl., Dimorphotheca pluvialis (L.)
Moench, Felicia tenella (L.) Nees., Steirodiscus tagetes (L.)
Schlechter, Tripteris clandestina Less., and Bellis perennis
L. Also, we did not observe three-celled spores in the
holotype (on Lagenophora billardieri). On Senecio spp.,
this feature is especially variable: 0% in KR 3073a, KR
8480 (S. vulgaris L.), KR 8468 (Senecio spec.), KR 8467
(S. inaequidens); 1-5% in KR 8481, KR 8483, KR 22642
(S. vulgaris), KR 20286 (S. vulgaris ssp. denticulatus); and
10-11% in KR 8500, KR 13384 (S. vulgaris). Of the two P.
saccardoi on Goodeniaceae, one had a high proportion of
three-celled spores (12%, VPRI 3906) whereas the other
had none at all (VPRI 3896).
Ridges on teliospores were observed in many specimens,
usually on less than 5% of spores but occasionally as much
as on approximately 20% of spores. In the holotype of P.
lagenophorae ridges were observed on one meso- and one
teliospore of 50 spores. One specimen of P. saccardoi
(VPRI 3906) contains an especially high proportion of
teliospores with ridges.
Phylogenetic analyses
All analysed LSU sequences were almost identical within
P. lagenophorae (ml1201 differed in position 91 of the
alignment; ml1279 in position 202; ml866, ml988 (= ml871),
and ml1006 in position 473; AY808061 in position 476).
The sequence of Aecidium sp. on Ammobium alatum R. Br.
differed in two positions, and that of the P. obscura
specimens in nine positions compared with those of P.
lagenophorae. ITS sequences were somewhat more variable.
Within the P. lagenophorae complex, seven different ITS-
5–10
4.5–12
4.5–10.5
6–10.5
7–10
a
ab
ab
bc
cd
cd
cd
cd
de
de
ef
f
0.9
1.1
1.29
1.13
1.05
0.98
1.62
1.04
1.08
genotypes were revealed which differed in up to 4 bp in a
total of six positions of the alignment: genotype 1:
AF468041, AY808060, EF212447, ml1011, ml1277;
genotype 2: ml1139; genotype 3: AF468040, EU391656,
ml868, ml870, ml1274; genotype 4: AY852264,
EF212446, EF635886, EF635889, EF635890, ml866,
ml867, ml872, ml1005, ml1006, ml1012; genotype 5:
EF635887; genotype 6: EF635888, ml869; genotype 7:
EF635891, EF635892, EF635893, EF635894. The genotypes are denoted in Fig. 1.
Different runs of Bayesian phylogenetic analyses and
the neighbour-joining analysis yielded consistent topologies. We present the consensus tree of one run of
Bayesian phylogenetic analyses to illustrate the results
(Fig. 1). Estimates for a posteriori probabilities are
indicated on branches before the slash, bootstrap values
from the neighbour-joining analysis after the slash. The
analyses revealed a well-supported cluster (a posteriori
probability: 97; bootstrap value: 95) of P. lagenophorae
including P. distincta sensu Weber et al. (1998). Two
additional species, P. saccardoi and P. stylidii, clustered
within the P. lagenophorae group, the former was the only
representative of genotype 5. Aecidium sp. on A. alatum
clustered as a well-supported sister taxon of the P.
lagenophorae complex. P. obscura and P. vaginatae are
more distantly related to the specimens studied here. P.
dioicae and A. brachycomes on Brachycome spathulata
Gaudich. were shown to be more closely related to the P.
lagenophorae complex than the previous species and “P.
lagenophorae” on Senecio madagascariensis Poir. from
South Africa (EF 635899, 635900). Compared to the
sequences of the P. lagenophorae complex, P. stylidii differed
in 10 bp, Aecidium sp. on A. alatum in 18 bp, and P. obscura
in 53 bp.
Mycol Progress (2011) 10:175–187
181
P. l. (EU391656: P. distincta) on Bellis perennis/Astereae (Denmark) - gt 3
P. l. (EF635894) on S. madagascariensis/Senecioneae (South Africa) - gt 7
P. l. (EF635893) on S. madagascariensis/Senecioneae (South Africa) - gt 7
P. l. (EF635892) on S. madagascariensis/Senecioneae (South Africa) - gt 7
P. l. (EF635891) on S. madagascariensis/Senecioneae (South Africa) - gt 7
P. l. (EF635888) on S. madagascariensis/Senecioneae (South Africa) - gt 6
70/- P. l. (EF635887: P. saccardoi) on G. hederacea ssp. alpestris/Asterales: Goodeniaceae (Australia) - gt 5
P. stylidii (EF635895) on Stylidium sp./Asterales: Stylidiaceae (Australia)
P. l. (AF468040: P. distincta) on Bellis perennis/Astereae (UK) - gt 3
P. l. (ml869) on Dimorphotheca pluvialis/Calenduleae (South Africa) - gt 6
P. l. (ml1274) on Bellis perennis/Astereae (New Zealand) - gt 3
P. l. (ml1139) on Bellis perennis/Astereae (Germany) - gt 2
P. l. (ml868) on Bellis perennis/Astereae (Germany) - gt 3
P. l. (ml870) on Bellis perennis/Astereae (Germany) - gt 3
P. l. (EF635890) on S. madagascariensis/Senecioneae (South Africa) - gt 4
P. l. (EF635889) on S. madagascariensis/Senecioneae (South Africa) - gt 4
P. l. (EF635886) on Arctotheca populifolia/Arctotideae (Australia) - gt 4
97/95
P. l. (EF212447) on S. vulgaris/Senecioneae (Canada) - gt 1
P. l. (EF212446) on S. vulgaris/Senecioneae (Canada) - gt 4
P. l. (AY808060) on Emilia sp./Senecioneae (UK) - gt 1
P. l. (AY852264) on S. vulgaris/Senecioneae (USA) - gt 4
P. l. (AF468041) on S. vulgaris/Senecioneae (UK) - gt 1
P. l. (ml866) on Dimorphotheca sinuata/Calenduleae (South Africa) - gt 4
100/94
P. l. (ml1006) on Gazania krebsiana/Arctotideae (South Africa) - gt 4
P. l. (ml1005) on Felicia tenella/Astereae (South Africa) - gt 4
P. l. (ml1011) on Osteospermum pinnatum/Calenduleae (South Africa) - gt 1
P. l. (ml1012) on S. arenarius/Senecioneae (South Africa) - gt 4
P. l. (ml1277) on S. vulgaris/Senecioneae (New Zealand) - gt 1
100/85
P. l. (ml867) on Bellis perennis/Astereae (USA) - gt 4
P. l. (ml872) on S. vulgaris/Senecioneae (USA) - gt 4
Aecidium sp. (ml1280) on Ammobium alatum/Gnaphalieae (Australia)
79/65
P. dioicae (EF635897) on Carex bichenoviana
95/55
Aecidium brachyscomes (EF635896) on Brachyscome spathulata
100/100 P. vaginatae (EF635902) on Saussurea alpina
P. vaginatae (EF635901) on Carex vaginata
55/61
100/100
P. obscura (AF468042) on Bellis perennis/Astereae (UK)
P. obscura (ml1137) on Luzula sylvatica (Germany)
P. obscura (ml1140) on Luzula campestris (UK)
100/99
P. l. (EF635900) on S. madagascariensis/Senecioneae (South Africa)
P. l. (EF635899) on S. madagascariensis/Senecioneae (South Africa)
P. rupestris (EF635898) on Carex rupestris
1 substitution/site
Fig. 1 Bayesian inference of phylogenetic relationships within the
sampled rust specimens: Markov chain Monte Carlo analysis of an
alignment of ITS base sequences using the GTR+I+G model of DNA
substitution with gamma distributed substitution rates and estimation
of invariant sites, random starting trees and default starting parameters
of the DNA substitution model. A 50% majority-rule consensus tree
computed from 16,000 trees that were sampled after the process had
reached stationarity is shown. The topology was rooted with Puccinia
rupestris (EF635898). Numbers on branches before slashes are
estimates for a posteriori probabilities, numbers on branches after
slashes are percentage bootstrap values of 1,000 replicates. Branch
lengths were averaged over the sampled trees. They are scaled in
terms of expected numbers of nucleotide substitutions per site. P.
Puccinia, S. Senecio, gt genotype (see the text)
Taxonomy
as well as on molecular data. Four taxa (P. brachycomes, P.
calotidis McAlpine, P. podolepidis, and P. vittadiniae
McAlpine) are listed as synonyms based only on morphological descriptions. Names listed here as synonyms for the
first time are given in bold type.
Puccinia lagenophorae Cooke, Grevillea 13 (65): 6
(1884) (Fig. 2)
Neither molecular nor morphological data support the split
of P. lagenophorae into different species. The following list
of synonyms is based on morphological and biological
studies of Cunningham 1931; Viennot-Bourgin 1964;
Wilson et al. 1965; Scholler 1993, 1997, and other authors
182
Fig. 2 The holotype of Puccinia lagenophorae (K(M)162111)
(host Lagenophora billardierei Cass., Asteraceae; holotype in K).
≡ Dicaeoma lagenophorae (Cooke) Kuntze, Revis. Gen.
Pl. (Leipzig) 3: 469 (1898)
= Puccinia saccardoi F. Ludw., Hedwigia 28: 362
(1889) (host Goodenia geniculata R. Br., Goodeniaceae)
≡ Dicaeoma saccardoi (F. Ludw.) Kuntze, Revis. Gen.
Pl. (Leipzig) 3: 470 (1898)
= Puccinia erechtitis McAlpine, Proc. Roy. Soc. Victoria
7: 216 (1894) (issued January 1895) (host Senecio quadridentatus Labill., originally as 'Erechtites quadridentata?',
Asteraceae)
= Puccinia hypochaeridis McAlpine (as 'hypochaeris'),
Proc. Roy. Soc. Victoria 7: 217 (1894) (issued January
1895) (host Lagenophora billardierei Cass., Asteraceae,
originally in error as Hypochaeris radicata), nom. illegit.
Art. 53 ICBN [non Puccinia hypochaeridis Oudem. 1873],
= Puccinia distincta McAlpine, Agric. Gaz. N.S.W., 6:
853 (1895) (issued 1896) (host Bellis perennis L., Asteraceae)
≡ Lindrothia distincta (McAlpine) Syd., Annal. Mycol.
20(3): 119 (1922)
= Puccinia nigricaulis McAlpine, Agric. Gaz. NSW 7:
151 (1896) (hosts Goodenia glauca F. Muell. and Goodenia
pinnatifida Schltdl., Goodeniaceae, a synonym of Puccinia
saccardoi according to McAlpine 1906)
= Puccinia macalpinei P. Syd. & Syd. (as 'macalpini'),
Monographia Uredinearum I: 100, (1902) (host Lageno-
Mycol Progress (2011) 10:175–187
phora billardierei Cass., Asteraceae, given originally in
error as 'Hypochoeris radicata'; nom. nov. for P. hypochaeridis McAlpine q.v.).
= Puccinia calendulae McAlpine, Proc. Linn. Soc. N.S.W.
28: 558 (1903) (host Calendula officinalis L., Asteraceae).
Anam.: Aecidium calendulae McAlpine, Agric. Gaz. N.
S.W. 7: 152 (1896)
≡ Lindrothia calendulae (McAlpine) Syd., Annal.
Mycol. 20(3): 119 (1922)
= Puccinia tasmanica Dietel, Ann. Mycol. 1: 535 (1903)
(host Senecio vulgaris L., Asteraceae)
≡ Lindrothia tasmanica (Dietel) Syd., Ann. Mycol. 20
(3): 119 (1922)
= Puccinia brachycomes McAlpine, Rusts of Australia:
150 (1906) (hosts Brachyscome ciliaris (Labill.) Less., B.
diversifolia (Hook.) Fisch. & C. A. Mey, B. pachyptera
Turcz., B. scapiformis DC., Asteraceae)
= Puccinia calocephali McAlpine, The Rusts of Australia: 151 (1906) (hosts Calocephalus drummondii (A.
Gray) Benth., C. lacteus Less., Asteraceae, a synonym of P.
erechtitis according to Cunningham 1931)
= Puccinia calotidis McAlpine, Rusts of Australia: 152
(1906) (hosts Calotis cuneifolia R. Br., Calotis sp.,
Asteraceae)
≡ Lindrothia calotidis (McAlpine) Syd., Ann. Mycol. 20
(3): 119 (1922)
= Puccinia cinerariae McAlpine, The Rusts of Australia:
155 (1906) (host Cineraria sp. (cult.), Asteraceae)
= Puccinia podolepidis McAlpine, Rusts of Australia:
162 (1906) (host Podolepis longipedata A. Cunn. ex DC.,
Asteraceae)
≡ Lindrothia podolepidis (McAlpine) Syd., Ann. Mycol.
20(3): 119 (1922)
= Puccinia vittadiniae McAlpine, Rusts of Australia:
164 (1906) (host Vittadinia australis A. Rich., Asteraceae)
= Puccinia cruciferae McAlpine, Rusts of Australia:
184 (1906) (host Vittadinia triloba (Gaudich.) DC.,
Asteraceae, originally in error as 'cruciferous plant'; see
Walker 1983)
= Puccinia allanii G. Cunn., Trans. Proc. N. Z. Inst. 54:
675 (1923) (host Senecio lagopus Raoul, Asteraceae).
= Puccinia byliana Dippenaar, S. Afr. J. Sci. 28: 288
(1931) (as ‘P. bylianum’, name corrected in Doidge 1950).
(on Senecio littoreus Thunb., S. pinnulatus Thunb, S.
vulgaris L. and Senecio sp., Asteraceae; Lectotype here
designated: South Africa, Western Cape, Hopefield, on
Senecio littoreus Thunb., IX 1925, leg. P.A. van der Byl
2356 (PREM 46293).
Anam.: Aecidium bylianum Syd., Ann. Mycol. 22: 236
(1924).
= Puccinia terrieriana Mayor, Ber. schweiz. bot. Ges.
72: 266 (1962) (type host Senecio vulgaris L., Asteraceae)
Mycol Progress (2011) 10:175–187
Host range
A total of 41 new host records are recorded herein, 38 from
South Africa, and 1 each from Germany, New Zealand and
Spain (for details, see Appendix A).
Discussion
Nomenclature, morphology, taxonomy, phylogeny
Our molecular analyses using ITS sequence data indicate
that Puccinia lagenophorae forms a well-supported cluster
with almost identical ITS sequences differing only in few
base pairs (Fig. 1), confirming Morin et al. (2009). This
cluster also includes the specimens on B. perennis which
were separated by Weber et al. (1998) as P. distincta. The
original sequences used by Weber et al. (2003) occur in two
sub-clades, but there is very low support for this separation.
Weber et al. (2003) only analysed material from western
Europe, where the rust is introduced, whereas here specimens were included from throughout the geographical
range, including Australia where the species originated.
Because of the unique ontogeny (autopsis-form with
repeating aecia, leptosporic teliospores), specific morphological features (e.g. aeciospores with hyaline plugs,
formation of mesospores), and host range (Asteraceae and
related Goodeniaceae) of taxa within this cluster, we
consider this cluster to represent one species. Outside this
cluster and not included in P. lagenophorae are two
specimens of “P. lagenophorae” on S. madagascariensis
(EF635899 and EF635900). Morin et al. (2009) provided
evidence that these two sequences resulted from an
interspecific hybridisation event, with P. lagenophorae as
one of its host parents and showing P. lagenophorae
morphology.
This lumping of rust species contrasts with several other
polyphagous “morphospecies”, for example the hostalternating Poaceae rusts P. graminis Pers. and P. coronata
Corda. Both these species are genetically diverse (Abbasi et
al. 2005; Szabo 2006) and probably need to be split into
numerous species. In addition, we include a nonasteraceous species, P. saccardoi, in P. lagenophorae. Its
host plants of the genera Goodenia, Scaevola and Velleia
belong to the Goodeniaceae, a family closely related to the
Asteraceae (Hansen 1997; Stevens 2001). Because of this,
P. lagenophorae is considered a polyphagous species in an
even broader sense than proposed by Wilson et al. (1965).
Several other species from Australia may also belong to this
broad concept of P. lagenophorae, including P. angustifoliae McAlpine (on Asteraceae) and P. dampierae P. Syd. &
Syd. (on Goodeniaceae). Before synonymising these spe-
183
cies, however, molecular and further morphological studies
are required. Puccinia stylidii, another non-asteraceous
species on Stylidiaceae (also related to Asteraceae; Stevens
2001), clustered within P. lagenophorae. However, the
genetic distance of P. stylidii compared the P. lagenophorae
sequences is quite considerable (10 base pairs in the ITS).
Its teliospores resemble P. lagenophorae, but it forms
uredinia and not aecia (McAlpine 1906). This indicates
that P. stylidii is a well-characterised species which likely is
derived from the same or similar parental species as P.
lagenophorae.
In Europe, specimens on B. perennis were regarded as a
distinct species from those on S. vulgaris because crossinoculations were unsuccessful and no pycnia were formed,
whereas the Senecio rust sometimes formed pycnia, the
teliospore pedicel width was smaller in the Bellis rust, and,
finally, the Bellis rust had only two-celled and one-celled
teliospores (mesopores), whereas the Senecio rust had in
addition three-celled teliospores (Weber et al. 1998; Jurc
and Weber 2000). Weber et al. (1998) speculated that the
Bellis rust derives from the European host-alternating P.
obscura (with pycnia and aecia on Bellis, and uredinia and
telia on Luzula). Later, Weber et al. (2003) provided a
molecular analysis, in which they regarded both the Bellis
and the Senecio rust as derived from P. obscura. However,
earlier authors made successful cross-inoculations from
Bellis to Senecio and vice versa (Viennot-Bourgin 1964;
Wilson et al. 1965). Recently, successful cross-inoculations
were carried out in North America by Koike and Scholler
(2001) and by Bruckart (2003); the latter author also used
inoculum from P. lagenophorae on S. vulgaris from Europe
(inoculum collected by C. Ellison, CABI, in March 1999,
from S. vulgaris, England; W. Bruckart, personal communivation). Also, successful cross-inoculations were carried
out in Germany by two of us (M.S. and G.H., unpublished
data, voucher specimens in herbarium KR).
Generally, we confirm the morphological description as
provided by Wilson et al. (1965). We could not confirm the
presence of pycnia as observed by McAlpine (1906),
Cunningham (1931), and Weber et al. (1998). On the
original material from New Zealand cited by Cunningham
(1931) on Lagenophora pumila Cheeseman (PDD 3217,
PDD 3784) and S. vulgaris (PDD 181, PDD 3543), no
pycnia were observed. On one specimen (PDD 1571),
pycnia were found (together with aecia and telia); however,
a detailed study indicated that this specimen is likely
misidentified. It differs from typical P. lagenophorae by
colour of sori and teliospore cell wall, size of teliospores
and by not forming mesospores. In addition, material
collected by R.W.S. Weber and J. Webster (K (M) 92025,
102618) was examined and no pycnia were observed. No
pycnia were observed on any other specimens examined.
184
The formation of pycnia in P. lagenophorae, if formed at
all, is rare, and it is possible the records are rather of
premature aecia which may have been misidentified as
pycnia, or that other pycnia-forming species may have been
misdentified as P. lagenophorae. Homothalism is common
amongst rust fungi with reduced life cycles, and is often
accompanied by the lack of pycnia (Ono 2002). The
teliospore pedicel width at the point of attachment to the
teliospore may depend on the host plant species and the part
of the plant where sori are formed. The pedicel width of the
type of P. lagenophorae is about the same as that given as
characteristic for P. distincta by Weber et al. (1998) on B.
perennis. It was found that this characteristic was highly
variable between specimens, with one specimen on A.
asteroides not only encompassing both ranges given by
Weber et al. (1998) but also having greater extreme values.
That P. lagenophorae on S. vulgaris consistently has a
broader point of attachment than on other hosts is
confirmed herein. P. distincta as defined by Weber et al.
(1998) cannot be distinguished from P. lagenophorae.
Although European specimens on B. perennis and S.
vulgaris were significantly different in pedicel width in
our analysis, other specimens were intermediate, and
therefore the differences observed by Weber et al. (1998)
represent two extremes of a continuum (Table 2). Threecelled teliospores are recorded as occasional in S. vulgaris
(Weber et al. 1998; Littlefield et al. 2005), and also in
Emilia spp. (Henricot and Denton 2005). In a specimen on
S. vulgaris from New York (KR 8483), only eight threecelled teliospores were found in 4 out of 10 sori observed
(400 teliospores observed in total). One of the two specimens on Goodeniaceae (VPRI 3906, formerly P. saccardoi
F. Ludw.) contains a high proportion of three-celled spores
whereas in the other one (VPRI 3896), not a single one was
observed. Three-celled teliospores were not observed in B.
perennis (Weber et al. 1998) and Senecio glaucus L. (Baka
1996), in the type on L. billardieri and on several hosts
from South Africa. Ridges on the teliospore surface were
illustrated by Viennot-Bourgin (1964, Fig. 2), Scholler
(1993), Wilson et al. (1965, Figs. 2, 11) and Wilson and
Henderson (1966). Other authors, however, such as Baka
(1996) and Cunningham (1931) do not mention them.
McAlpine (1906) does not mention them either although
these can be seen in photographs of teliospores of P.
calendulae and P. brachycomes (p. 256). Ridges were
observed in the majority of specimens studied, and their
occurrence was independent of the host plant and geographical origin. Therefore, we do not agree with Weber et
al. (1998) who consider ridges to be an artifact in fresh
spores mounted in lactic acid (the photograph in Weber et
al. 1998, Fig. 8, indeed does not show ridges but shrunken
spores, possibly as a consequence of insufficient heating of
lactic acid microscopic mounts). According to J. Walker
Mycol Progress (2011) 10:175–187
(personal communication), ridges may be the result of
mutual pressure between the spores in the telia rather than
true ornamentation. This is supported by the fact that the
ridges were observed to mostly have formed in regions of
the spore where the wall is thin. Our ontogenetical and
morphological studies and those of other authors mentioned
above have shown that the formation or non-formation of
pycnia, three-celled teliospores and the width of the pedicel
are very variable and not constant characters of P.
lagenophorae. These characters may, to some degree, be
influenced by the morphology of the host species, but they
are not useful taxonomic characteristic as claimed by Weber
et al. (1998).
The results do not support the suggestion made by
Weber et al. (2003) that P. obscura may be the parental
species of P. lagenophorae. The molecular analyses show at
least three species (A. brachycomes, Aecidium sp. and P.
dioicae) being more closely related to P. lagenophorae than
P. obscura, Morin et al. (2009) previously also obtained the
same result. The same may hold for P. vaginatae, although
support values are low. But we support Weber et al. (2003)
that the parental species is likely to be, like P. obscura, a
host-alternating species between Asteraceae (aecial host)
and Cyperaceae/Juncaceae (telial host). Puccinia dioicae
(and P. vaginatae) have such a life cycle and Aecidium sp.
and A. brachycomes (both on Asteraceae) may also be part
of the life cycle of a host-alternating species with a
Cyperaceae/Juncaceae telial host.
Host range
Based on our species concept and considering the large
number of host species from South Africa, the number of
known hosts is increased from about 60 to 150, about 40 of
which are members of the Goodeniaceae. In contrast to
Wilson et al. (1965) and Scholler (1998), who listed only
species of the subfamily Asterioideae, we found that
members of the subfamily Cichorioideae (Arctotis, Arctotheca, Gazania, Vernonia) and of the family Goodeniaceae
(Goodenia, Scaevola, Selliera, Velleia) also host P. lagenophorae. Since the host range of rust fungi is mostly
restricted to groups of related plant species, P. lagenophorae may provide further information for plant systematists on relationships between the Asteraceae and related
families.
Annotations to geographic origin and pathways
of expansion
Wilson et al. (1965) were the first to name P. lagenophorae
as a rust originating from Australasia (Australia, New
Zealand, New Guinea) which had been introduced to
Europe. Further arguments in support of this statement
Mycol Progress (2011) 10:175–187
are: the first record (the holotype) is from Australia, the
highest number of host plants is recorded in Australasia,
there are also other related rust species with hosts on
Asteraceae or related families with similar teliospore and
mesospore morphology (e.g. P. brunoniae McAlpine, P.
gilgiana Henn. on Goodeniaceae, and P. stylidii on
Stylidaceae), and similar species are not known from
elsewhere. Finally, our molecular studies indicate that there
is the highest (relative) genetic diversity of P. lagenophorae
in Australasia with four different ITS-genotypes in four
specimens (the quotient of the number of genotypes :
number of samples is 1 for Australasia, 0,43 for Europe, 0,4
for North America, and 0,33 for South Africa). Highest
genetic diversity is a feature typical for the place of origin
of fungi (e.g. Zhang et al. 2009).
The earliest record of P. lagenophorae traced for Africa
was from the Western Cape Province in South Africa
collected in 1918 (see Appendix A). So far P. lagenophorae
has not been recorded from elsewhere in sub-saharan
Africa. However, a record from Sierra Leone from 1930
was discovered in PUR (see Appendix A). Furthermore
there is a single record of an unidentified Aecidium on an
ornamental Cineraria sp. from Tanzania (Wallace 1936)
which may represent this species. This raises the possibility
that this fungus may have reached Europe from Africa (as
previously suggested by Viennot-Bourgin 1964) and not
necessarily directly from Australasia.
The molecular data presented herein do not provide
information about the pathways of expansion of P.
lagenophorae as too few isolates were included for this
purpose, especially from Australasia. In each geographic
region (Australasia, Europe, North America, South Africa),
we found at least two different ITS genotypes (although
only differing in few base pairs). It is interesting to note that
the most common genotype (genotype 4) occurred in
specimens from Australia, North America, and South
Africa but not Europe. Also, genotype 3 was only recorded
from specimens on B. perennis from Europe and New
Zealand, yet the two genotypes that were most similar were
only recorded from South Africa (genotypes 6 and 7).
There is thus the suggestion that multiple inter-continental
introductions have been made.
Practical aspects
Several host plants of P. lagenophorae are important
ornamental plants. In Europe these include Asteriscus
maritimus (L.) Less., Calendula officinalis L., Dimorphotheca pluvialis (L.) Moench and D. sinuata DC., Emilia
coccinea (Sims) G. Don and E. sonchifolia (L.) DC.
Senecio glaucus L. subsp. coronopifolius (Maire) Alex.,
and Pericallis X hybrida B. Nord. (S. cruentus DC.,
Pericallis cruenta (L'Hér. ) Bolle), Bellis perennis is a wild
185
plant in Europe but certain cultivars are attractive ornamental plants and rust infection causes economic losses. As
a consequence, plant pathologists in various European
countries developed and tested fungicides to control the
daisy rust (e.g. Gullino et al. 1999; Weber and Tilston 1999;
Gerlach 2000). On the other hand, numerous efforts were
made to develop P. lagenophorae as a biocontrol agent
against the weed S. vulgaris (e.g. Müller-Schärer and
Rieger 1998; Killgore et al. 2000; Frantzen et al. 2002;
Bruckart 2003; Frantzen and Müller-Schärer 2006).
In this context it is noteworthy that previous authors like
Viennot-Bourgin (1964), Wilson et al. (1965), Koike and
Scholler (2001), and Bruckart (2003) were successful in
making cross-inoculations with P. lagenophorae on Bellis
and Senecio. The reason why Weber et al. (1998) failed
remains unknown. Wilson et al. (1965) made successful
cross-inoculations from S. vulgaris to B. perennis, Pericallis X hybrida B. Nord. and C. officinalis in the
greenhouse, but did not observe this rust on B. perennis
in the field. Cross-infection between Bellis and Senecio and
vice versa, however, probably often occurs even in nature
as suggested by floristic data. In Germany (Scholler 1994),
Poland (Piatek 2003), and California (Scholler and Koike
2001; Koike and Scholler 2001), for instance, the first
records on B. perennis and S. vulgaris were found in the
same area outside greenhouses. Furthermore, such records
were either found in the same year (in Germany in 1966,
in Poland in 1998) or only one year later after direct
searching (California). There are few data available to
determine whether races occur within P. lagenophorae or
not, and if so what are their respective host ranges. If such
races do occur, their host range may be broad, as
inoculation experiments have shown that inoculum
derived from Senecio (Asterioideae) can infect Vernonia
(Cichorioidae) (Table 1). The consequences of this finding
of polyphagy in P. lagenophorae may affect the research
of plant pathologists and ecologists throughout its distribution range. Our study confirms the finding by Wilson et
al. (1965) that P. lagenophorae is polyphagous and
therefore cognizance should be made as to whether any
potential conflicts of interest occur where use is made of it
as a biocontrol agent. It is recommended that Common
Groundsel and other potential host plants in the vicinity of
English Daisy plantings are removed as a potential control
measure.
Acknowledgements We thank John Walker for checking the list of
synonyms, Walter Gams for reviewing an earlier version of the
manuscript, Bill Bruckart for providing methodical information, Ralf
Jahn for identifying Senecio rodriguezzii and the curators of Herbaria
BOL, DAR, K, NBG, PDD, PUR, PREM, SAM and VPRI for
providing specimens of Puccinia lagenophorae and related species.
Field studies carried out by M. Menniken in South Africa were
financed by a grant from BMBF, Germany (project “BIOTA”).
186
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