S
choller
et al.: Erysiphe platani – monitoring of an epidemic spread in Germany
265
in order to elucidate the phylogenetic placement
and compare sequence data from specimens of
different geographic origin.
2 Materials and methods
2.1 Documentation of the epidemic spread
in Germany
After finding Erysiphe platani at a parking lot in
Tul (Lorraine, France) in 2008 the first author
looked methodically for this fungus in the Karls
ruhe region (Baden-Württemberg, Germany).
After finding it in August 2009 a call to mycolo-
gists and botanists was started in September
2009 to look for this fungus and help to document
its spread in Germany. Collaborators were asked
via email to provide both positive and negative
record data and voucher specimens. They were
regularly informed about the present distribu-
tion of the fungus in order to look for it where
it was still missing. Voucher specimens were all
checked by the first author and finally deposited
in the fungus herbarium of the Natural History
Museum in Karlsruhe (KR). A few specimens are
also deposited in the fungus herbarium in Görlitz
(GLM). All specimen data including information
on anamorph and teleomorph formation, hyper-
parasites etc. will be available via diversity collec-
tion and IMDAS online databases. The following
is a list of collaborators (authors not mentioned):
H
erbert
B
oyle
, A
drien
B
olay
, U
we
B
raun
, W
alter
G
ams
, P
atrick
D
ornes
, H
orst
J
age
, R
oland
K
ir
schner
, F
riedemann
K
lenke
, H
anns
K
reisel
, J
ulia
K
ruse
, V
olker
K
ummer
, D
irk
M
atalla
, B
ertold
M
etzler
, B
ernd
O
ertel
, H
arald
O
strow
, M
arçin
P
iatek
, U
do
R
ichter
, H
arry
R
egin
, A
nnemarthe
R
ubner
, A
nke
S
chmidt
, M
artin
S
chnittler
, D
iet
rich
S
choller
, K
atharina
S
choller
, M
anfred
S
chubert
, L
eopold
S
chrimpl
, H
orst
S
taub
, and
H
jalmar
T
hiel
.
2.2 Molecular analyses
DNA extraction, PCR, and sequencing
The specimens examined in the course of this
study are listed in Table 1. The voucher speci-
mens are deposited in KR. Genomic DNA was
isolated directly from the herbarium specimens.
For methods of isolation and crushing of fungal
material, DNA extraction, amplification, purifica-
tion of PCR products, sequencing, and proces-
sing of the raw data see L
utz
et al. (2004). ITS 1
and ITS 2 regions of the rDNA including the 5.8S
rDNA (ITS) were amplified using the primer pair
ITS1-F (G
ardes
& B
runs
1993) and ITS4 (W
hite
et al. 1990). The 5´-end of the nuclear large sub-
unit ribosomal DNA (LSU) was amplified using
the primer pair NL1 and NL4 (O´D
onnell
1993).
Primers were used for both PCR and cycle se-
quencing. For amplification the annealing tempe-
rature was adjusted to 45° C. DNA sequences
determined in this study were deposited in Gen-
Bank. GenBank accession numbers are given in
Fig. 2 and Table 1.
Phylogenetic analyses
The Erysiphe specimens examined in this study
are listed in Table 1. For molecular phylogenet-
ic analyses the following ITS sequences from
GenBank were additionally used (A
ttanayake
et al. 2010; B
raun
et al. 2006; C
ook
et al. 2004,
2006; C
unnington
et al. 2003; F
rancis
et al. 2007;
H
eluta
et al. 2009; H
irata
& T
akamatsu
1996; K
ho
daparast
et al. 2003; K
irschner
2010; K
ovacs
et
al. 2011; L
ee
et al. 2011; L
imkaisang
et al. 2006;
M
ori
et al. 2000; S
aenz
& T
aylor
1999; S
eko
et
al. 2011; S
hiroya
& T
akamatsu
2009; S
tanosz
et
al. 2009; and T
akamatsu
et al. 1998, 1999, 2006,
2007, 2008): Erysiphe abbreviata (P
eck
) U. B
raun
& S. T
akam
. AB271785, E. adunca (W
allr
.) F
r
.
var. adunca D84382, E. alphitoides (G
riffon
&
M
aubl
.) U. B
raun
& S. T
akam
. AB292710, E. aqui-
legiae DC. HQ286643, E. arcuata U. B
raun
, V. P.
H
eluta
& S. T
akam
. AB252462, E. astragali DC.
AB104515, E. baeumleri (M
agnus
) U. B
raun
&
S. T
akam
. AB015933, E. betae (V
a
ň
ha
) W
eltzien
DQ164436, E. buhrii U. B
raun
AB128924, E. car-
pinicola (H
ara
) U. B
raun
& S. T
akam
. AB252469,
E. carpini-laxiflorae U. B
raun
, V. P. H
eluta
& S. T
a
kam
. AB252470, E. catalpae S
imonyan
DQ359695,
E. circaeae L. J
unell
AB104517, E. clandestina
B
iv
. AB475115, E. convolvuli DC. AF011298,
E. corylopsidis S
hiroya
& S. T
akam
. AB478990,
E. cruciferarum O
piz
ex
L. J
unell
EU140958,
E. densa B
erk
. DQ005439, E. deutziae (B
unki
na
) U. B
raun
& S. T
akam
. GU196146, E. eleva-
ta (B
urrill
) U. B
raun
& S. T
akam
. AY587013, E.
euonymi-japonici (V
ienn
.-B
ourg
.) U. B
raun
& S.
T
akam
. AB250228, E. fimbriata S. T
akam
., M
asu
ya
& Y. N
omura
AB333839, E. friesii var. dahuri-
ca (U. B
raun
) U. B
raun
& S. T
akam
. AB000939;
E. glycines F. L. T
ai
var. glycines AB015934, E.
glycines var. lespedezae (R.Y. Z
heng
& U. B
raun
)
U. B
raun
& R. Y. Z
heng
AB015923, E. helwingi-
ae (S
awada
) U. B
raun
& S. T
akam
. AB015916, E.
heraclei DC. AB104510, E. howeana U. B
raun
AF011301, E. huayinenesis R. Y. Z
heng
& G.
Q. C
hen
AB015914, E. hypogena S. T
akam
. &
