Erysiphe Platani : Monitoring of an Epidemic Spread in Germany and Molecular Characterization Based on Rdna Sequence Data
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©Staatl. Mus. f. Naturkde Karlsruhe & Naturwiss. Ver. Karlsruhe e.V.; download unter www.zobodat.at SCHOLLER et al.: Erysiphe platani – monitoring of an epidemic spread in Germany 263 Erysiphe platani : monitoring of an epidemic spread in Germany and molecular characterization based on rDNA sequence data MARKU S SCHOLLER , VERENA HE mm & MATTHIA S LUTZ Abstract ist Arendsee im nördlichen Sachsen-Anhalt. Vermutet This work deals in two sections with the North Ameri- wird, dass die Art über das Rhonetal und die Burgun- can plane powdery mildew Erysiphe platani, an epide- dische Pforte in die Oberrheinebene nach Deutschland miological study and a molecular phylogenetic analysis eingedrungen ist. based on rDNA ITS sequence data. Most likely, the Die Sequenzanalyse zeigt, dass Material aus Deutsch- species was introduced in South Europe at the begin- land und Italien identisch ist und sich von Material aus ning of the 1960s. In 2007, it was observed for the first Griechenland leicht und deutlich von außereuropä- time in Germany near Freiburg (SW Germany) and ob- ischem Material (Australien, USA) unterscheidet. Dies viously did not reach other German states until 2009. A deutet auf eine erhebliche Mutationsrate im neuen eu- detailed monitoring from 2009 to 2011 shows that the ropäischen Areal hin. Die phylogenetische Auswertung fungus continually spread north- and northeastward der Sequenzdaten bestätigt, dass Erysiphe platani mit with a speed of roughly 190 km/year. The northernmost anderen auf Gehölzen vorkommenden Erysiphe-Arten record is from Arendsee in the north of Sachsen-Anhalt verwandt ist, die genau wie E. platani früher zur Gat- from 2011. We assume that the species has come from tung Microsphaera gestellt wurden. the Rhone valley and the Burgundian Gate finally en- tering Germany in the Upper Rhine plain. Autoren The molecular phylogenetic analyses of material of Dr. MARKU S SCHOLLER , Dipl.-Geoökol. VERENA HE mm , different geographic origins indicate that specimens Staatliches Museum für Naturkunde, Abt. Biowissen- from Germany and Italy are identical, differ slightly schaften, Erbprinzenstr. 13, 76133 Karlsruhe, E-Mail: from those from Greece and strongly from extraeuro- [email protected] pean (Australia, USA) material. This might indicate a Dr. MATTHIA S LUTZ , Universität Tübingen, Institut für considerable rate of mutation of this powdery mildew Evolution & Ökologie, Auf der Morgenstelle 1, 72076 with North American origin in the new European area. Tübingen In addition, the phylogenetic analyses confirm that E. platani is related to other tree-inhabiting powdery mil- dew species previously accommodated in the genus 1 Introduction Microsphaera. Powdery mildews (Erysiphales, Ascomycota) are Kurzfassung obligate plant-parasitic microfungi predominantly Erysiphe platani: Monitoring einer epidemischen on dicots forming white conspicuous “powdery” Ausbreitung in Deutschlandund molekulare Cha- conidial layers on plant surfaces, predominantly rakterisierung basierend auf rDNS-Sequenzdaten on leaves, which are often deformed (Fig. 1). Die vorliegende Arbeit fasst die Ergebnisse zweier Untersuchungen des aus Nordamerika stammenden Fruitbodies are black, globose, without opening Echten Mehltaupilzes der Platane, Erysiphe platani, (called cleistothecia or chasmothecia) and with zusammen: Eine epidemiologische Untersuchung und appendages. Conidia, ascospores and fruitbodies eine Sequenzanalyse von rDNS ITS-Sequenzen. Die may be dispersed by the wind, conidia in highest vermutlich Anfang der 1960er Jahre nach Südeuropa quantities (e.g. BLU M ER 1967). In an unpublished eingewanderte Art wurde 2007 erstmals in Deutsch- inventory, SCHOLLER et al. (2011) list 141 pow- land bei Freiburg (SW-Deutschland) nachgewiesen. dery mildew species for Germany. Fifty species, Für den Zeitraum von August 2009 bis 2011 wird die i.e. more than 1/3 of all species, are introduced epidemische Ausbreitung der Art innerhalb Deutsch- (so-called neomycetes and ephemeromycetes; lands dokumentiert. Von 2007 bis 2009 breitete sich Erysiphe platani nur langsam aus und erreichte ver- KREI S EL & SCHOLLER 1994) with a culmination of mutlich noch nicht die anderen Bundesländer. Danach introductions in the past two decades. Interest- beschleunigte sich die Ausbreitung auf rund 190 km/ ingly, many species have woody host plants. One Jahr und die Art breitete sich kontinuierlich nord- und of the most recently introduced species in Ger- nordostwärts aus. Der nördlichste Nachweis von 2011 many is the plane powdery mildew Erysiphe pla- ©Staatl. Mus. f. Naturkde Karlsruhe & Naturwiss. Ver. Karlsruhe e.V.; download unter www.zobodat.at 264 andrias, 19 (2012) Figure 1. The anamorph of the Plane Powdery Mildew Erysiphe platani on deformed leaves of Platanus acerifolia in Karlsruhe, Main Railway Station, August 27, 2009. – Photograph: M. SCHOLLER . tani (HOWE ) U. BRAUN & S. TAKA M . (= Microsphaera climates (AN S EL M I et al. 1994)1. Erysiphe platani platani HOWE ) with North American origin. The was reported on the host plants Platanus occi- species was recorded for the first time in Europe dentalis L., P. orientalis L., and the hybrid P. aceri- by CIFERRI & CA M ERA (1962) on Platanus orientalis folia (AITON ) WILLD . forming the anamorph only. In in Italy. However, the authors did not provide any the beginning of the 1990s the species appeared record data. Later GULLINO & RAPETTI (1978) found in south and southwestern Switzerland near lake the species in Liguria in Italy and documented Geneva (BOLAY 2005) indicating that the species its spread toward the French Mediterranean is spreading northward and may be expected in coast. In the following years the species spread Germany as well. Although there are several “first throughout the warm regions of the Mediterrane- record” publications from some countries with an countries, southern Switzerland, the mild at- morphological descriptions in Europe, little is lantic France, and southern UK but not to Central known about the details of the spreading mode, or Eastern Europe with increasingly continental e.g. speed of expansion and if progressing over long or short distances. This lack of information 1 In their distribution map AN S EL M I et al. (1994: 165, Fig. 3) does not concern only Erysiphe platani, but al- report of isolated records of Erysiphe platani in Southern most all other powdery mildew species intro- Sweden. Since the fungus was not observed in neighboring duced to Europe. In the following we present the countries we wanted to get this confirmed and asked the results of a monitoring of the epidemic spread authors directly, but did not get any response. The Swedish Erysiphales expert LENA JON S ELL could not confirm the occur- of E. platani in Germany. In addition to this, mo- rence of E. platani in Sweden. lecular phylogenetic analyses were carried out ©Staatl. Mus. f. Naturkde Karlsruhe & Naturwiss. Ver. Karlsruhe e.V.; download unter www.zobodat.at SCHOLLER et al.: Erysiphe platani – monitoring of an epidemic spread in Germany 265 in order to elucidate the phylogenetic placement ITS1-F (GARDE S & BRUN S 1993) and ITS4 (WHITE and compare sequence data from specimens of et al. 1990). The 5´-end of the nuclear large sub- different geographic origin. unit ribosomal DNA (LSU) was amplified using the primer pair NL1 and NL4 (O´DONNELL 1993). Primers were used for both PCR and cycle se- 2 Materials and methods quencing. For amplification the annealing tempe- rature was adjusted to 45° C. DNA sequences 2.1 Documentation of the epidemic spread determined in this study were deposited in Gen- in Germany Bank. GenBank accession numbers are given in After finding Erysiphe platani at a parking lot in Fig. 2 and Table 1. Tul (Lorraine, France) in 2008 the first author looked methodically for this fungus in the Karls- Phylogenetic analyses ruhe region (Baden-Württemberg, Germany). The Erysiphe specimens examined in this study After finding it in August 2009 a call to mycolo- are listed in Table 1. For molecular phylogenet- gists and botanists was started in September ic analyses the following ITS sequences from 2009 to look for this fungus and help to document GenBank were additionally used (ATTANAYAKE its spread in Germany. Collaborators were asked et al. 2010; BRAUN et al. 2006; COOK et al. 2004, via email to provide both positive and negative 2006; CUNNIN G TON et al. 2003; FRANCI S et al. 2007; record data and voucher specimens. They were HELUTA et al. 2009; HIRATA & TAKA M AT S U 1996; KHO - regularly informed about the present distribu- DAPARA S T et al. 2003; KIR S CHNER 2010; KO V AC S et tion of the fungus in order to look for it where al. 2011; LEE et al. 2011; LI M KAI S AN G et al. 2006; it was still missing. Voucher specimens were all MORI et al. 2000; SAENZ & TAYLOR 1999; SEKO et checked by the first author and finally deposited al. 2011; SHIROYA & TAKA M AT S U 2009; STANO S Z et in the fungus herbarium of the Natural History al. 2009; and TAKA M AT S U et al. 1998, 1999, 2006, Museum in Karlsruhe (KR). A few specimens are 2007, 2008): Erysiphe abbreviata (PECK ) U. BRAUN also deposited in the fungus herbarium in Görlitz & S. TAKA M . AB271785, E. adunca (WALLR .) FR. (GLM). All specimen data including information var. adunca D84382, E. alphitoides (GRIFFON & on anamorph and teleomorph formation, hyper- MAU B L .) U. BRAUN & S. TAKA M . AB292710, E. aqui- parasites etc. will be available via diversity collec- legiae DC. HQ286643, E. arcuata U. BRAUN , V. P. tion and IMDAS online databases. The following HELUTA & S. TAKA M . AB252462, E. astragali DC. is a list of collaborators (authors not mentioned): AB104515, E. baeumleri (MA G NU S ) U. BRAUN & HER B ERT BOYLE , ADRIEN BOLAY , UWE BRAUN , WALTER S. TAKA M . AB015933, E. betae (VAňHA ) WELTZIEN GA ms , PATRICK DORNE S , HOR S T JA G E , ROLAND KIR- DQ164436, E. buhrii U. BRAUN AB128924, E. car- S CHNER , FRIEDE M ANN KLENKE , HANN S KREI S EL , JULIA pinicola (HARA ) U.