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ZECH MYCOL . 62(1): 59–65, 2010

First record ofCeratocystis laricicola

(Ascomycota,Ceratocystidaceae) in the Czech Republic

DAVID NOVOTNÝ

Crop Research Institute, Division of Plant Medicine, Drnovská 507,

161 06 Praha 6 – Ruzyně, Czech Republic;
novotny@vurv.cz

Czech University of Life Science, Faculty of Forestry and Wood Science, Kamýcká 1176,

165 21 Praha 6 – Suchdol, Czech Republic

Novotný D. (2010): First record ofCeratocystis laricicola(Ascomycota,

Ceratocystidaceae) in the Czech Republic. – Czech Mycol. 62(1): 59–65.

During the study of large larch bark beetle (Ips cembrae)mycobiota, the speciesCeratocystis

laricicolawas isolated. It was detected in galleries and on the surface of the beetle body. This fungus is

a well-known larch pathogen and this observation is the first report of its occurrence from the Czech

Republic.

Key words:Ascomycota, ophiostomatoid fungi,Larix decidua, distribution,Ips cembra.

Novotný D. (2010):Ceratocystis laricicola(Ascomycota,Ceratocystidaceae),

první nález v České republice. – Czech Mycol. 62(1): 59–65.

Při studiu mykobioty lýkožrouta modřínového (Ips cembrae) byl z chodbiček a povrchu těla toho-

to brouka izolován druhCeratocystis laricicola, který je znám jako patogen modřínů. Jedná se o první

nález z České republiky.

INTRODUCTION

Ophiostomatoid fungi areOphiostoma–like genera usually associated with

bark beetles. The genusCeratocystis(Ceratocystidaceae, Microascales, Sorda-

riomycetes, Ascomycota) belongs to the best known taxa in this group.Ophio-

stomaandCeratocystisare not closely related; they belong to the ordersOphio-

stomatalesandMicroascales, respectively. They differ in anamorphs, cyclo-

heximide sensitivity, composition of cell wall and molecular-genetic features

(Kirk et al. 2001, Zipfel et al. 2006).

Ceratocystisspecies live more frequently in tropic regions than in the temper-

ate zone.Ceratocystis fagacearum(cause of oak wilt in North America),

C.fimbriata,C.coerulescens, andC.polonicaare the best known species living in

the temperate zone (Kile 1993, Marin et al. 2005).

Ceratocystis laricicolais associated with large larch bark beetle (Ips cembrae)

living on larch. It is very similar and related toC. polonica(living in connection

with bark beetles on spruce) and C.fujiensis, living in connection withIps sub

-

elongatusonLarix kaempferiin Japan.Ceratocystis laricicolaandC. polonica

are known pathogens of larch and spruce, respectively (Harrington et al. 2002,

Marin et al. 2005).

To the present day, twoCeratocystisspecies (C. autographaandC. polonica)

have been recorded in the Czech Republic.Ceratocystis autographawas ob

-

served by Kotýnková-Sychrová (1966) in connection with the bark beetleTomicus

piniperda. Ceratocystis polonicawas recorded repeatedly in the mycobiota of

Ips typographus(Jankovský and Mrkva 1997, Novotný and Jankovský 2005).

Until recently,Ceratocystis laricicolahad not been recorded in the Czech Re

-

public. This is the first report. The aim of this paper is to provide information on

the occurrence of this fungal species in the Czech Republic.

M

ATERIALS AND METHODS

During 2006, fungi living on adult beetles ofIps cembrae(Curculionidae,

Coleoptera) and in their galleries inLarix deciduawere investigated on Hudečka

hill near Stříbrná Skalice in the Sázava region in Central Bohemia, Czech Repub-

lic. Twenty beetles and samples of detritus from their galleries were separately in-

oculated on Petri dishes with 2 % malt extract agar. After 14–21 days of incubation

at room temperature the fungi were isolated and identified.

The isolated strain ofCeratocystis laricicolawas preserved under mineral oil

and deposited as CPPF 309 in the Collections of phytopathogenic microorganisms

(Crop Research Institute, Division of Plant Medicine, Department of Mycology,

Prague).

Growth of the isolated strain was tested on 2 % malt extract agar (MA2), po-

tato-dextrose agar (PDA), potato-carrot agar (PCA), oatmeal agar (OA) and 0.1 %

malt extract agar with a small piece of larch branch (MA01+LAR) at 20 °C (four

replicates in all media). The strain was cultivated at 32 °C, too.

The identification was based on morphological, physiological and ecological

features according to Redfern et al. (1987), Yamaoka et al. (1998), and Marin et al.

(2005).

The strainCeratocystis polonica(Siemaszko) C. Moreau CPPF 252 was used

for comparison. It was obtained from a gallery of the bark beetleIps typographus

in Southwest Moravia near Mohelno, Czech Republic.

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CZECH MYCOL . 62(1): 59–65, 2010

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NOVOTNÝ D.: F IRST RECORD OF CERATOCYSTIS LARICICOLAIN THE CZECH REPUBLIC

a b

c

Fig. 1.Ceratocystis laricicolaCPPF 309 –a:

ten-day old colony with arising perithecia on

MA01 + LAR at 20°C,b: ten-day old colony on

MA2 at 20°C – face,c: ten-day old colony on MA2

at 20°C – reverse,d: perithecium. Scale bar for a,

b, c = 20 mm, d = 11μm.

d

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CZECH MYCOL . 62(1): 59–65, 2010

a b

c

Fig. 2.Ceratocystis laricicolaCPPF 309 –a: ascospores,b: apex of neck with ostiolar hyphae,c:as-

cospores. Scale bar for a = 11μm, b = 12μm, c = 2μm.

RESULTS

Ceratocystis laricicolaRedfern et Minter 1987

The fungus occurred in 50 % of investigated beetle bodies and 55 % of the bee

-

tle’s galleries.

The description is based on a single representative strain: CPPF 309: gallery of

bark beetleIps cembrae, branch ofLarix decidua, slope of Hudečka hill near

Stříbrná Skalice, Sázava region, Central Bohemia, Czech Republic, isol. and det.

D. Novotný as no. IC15L, VI. 2006.

Only one strain was deposited and investigated, because the species was re

-

corded to be associated withIps cembraeon one tree at one locality only.

C o l o n y c h a r a c t e r i s t i c s (growth rates see in Tab. 1)

MA2, 10 days, 20 °C: colonies grey-white, flat, exudate absent, reverse grey

brown-green, soluble pigment absent.

OA, 10 days, 20 °C: colonies grey-white, flat, low, exudate absent, reverse pale

to greyish, soluble pigment absent.

PDA, 10 days, 20 °C: colonies grey-white, flat, exudate absent, reverse fuscous,

soluble pigment absent.

PCA, 10 days, 20 °C: colonies grey, flat, exudate absent, reverse grey-green, sol-

uble pigment absent.

MA01+LAR, 10 days, 20 °C: colonies dark brown-green, sparse, flat, very low,

exudate absent, reverse dark green in the middle, without colour at the margin,

soluble pigment absent.

MA2, 8 days, 32 °C: colonies grey-white, 29–32 mm in size, flat, brown, exudate

absent, reverse grey to brown-green.

The studied strain grows most quickly on PDA (see Tab. 1). The slowest growth

was recorded on MA2. Perithecia developed earliest and most abundant on

MA01+LAR medium.

Tab. 1.Growth ofCeratocystis laricicolaCPPF 309 on five different cultivation media at 20 °C.

Medium Colony diameter (mm)

5 days 7 days 10 days

MA2 16–23 20–35 43–64

PDA 23–32 49–54 78–85

OA 19–34 38–50 70–82

PCA 19–25 33–43 62–74

MA01+LAR 17–23 20–34 50–67

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NOVOTNÝ D.: F IRST RECORD OF CERATOCYSTIS LARICICOLAIN THE CZECH REPUBLIC

Microscopic features

Hyphae hyaline, light brown to dark brown, septate, (2–)2.5–3.0(–3.5)μm

wide, smooth. In culture, perithecia develop superficially within 10 days. Bases

globose, 160–240μm in diameter, dark brown or black. Necks straight or slightly

curved, brown to black, 580–890μm in length, 25–43μm wide at base, 14–23μm

wide at the tip immediately below the apex. Ostiolar hyphae present or absent in

early stages, hyaline, septate, convergent or slightly divergent, 18–50μm long and

2μm wide at base, tapering to apex. Asci not observed. Ascospores hyaline, one-

celled, oval, surrounded by mucous sheath, 5–6 × 2–2.5(–3)μm, emerging from

the ostiole and forming a whitish to yellowish droplet at the tip.

Anamorph:Thielaviopsiswas not observed on the isolated strain.

D

ISCUSSION

Ceratocystis laricicolaoccurs in connection withIps cembraein Europe

(Redfern et al. 1987, Marin et al. 2005, Jankowiak et al. 2007). So far,Ceratocystis

laricicolahas been isolated from larch in Poland (Jankowiak et al. 2007), Austria

(Marin et al. 2005) and United Kingdom (Redfern et al. 1987). It probably occurs in

other European countries whereIps cembraelives. The present article gives the

first report of the occurrence of this fungus from the Czech Republic. In JapanC.

fujiensisoccurs, which is very similar to this species. The description ofC.

laricicolagiven by Yamaoka et al. (1998) is in reality a description ofC.fujiensis,

as concluded by Marin et al. (2005), who revised some strains ofC. laricicolade-

scribed by Yamaoka et al. (1998) and identified them asC.fujiensis.Marin et al.

(2005) recorded onlyC.fujiensisin Japan. They did not obtain any strain ofC.

laricicolafrom Japan.

C.laricicola,C. polonica, andC.fujiensisare very closely related but differ in

growth rate at 32 °C, sequences ofβ-tubulin and MAT-2 HMG box and ecological

niche. On 2 % malt extract agar after 8 days at 32 °C, colonies ofC.laricicolaare

11–29 mm in diameter, whereas colonies ofC.polonicaunder the same conditions

do not exceed a diameter of 0–3 mm.C.fujiensisoccurs in connection withIps

cembraeliving onLarix kaempferionly.C.laricicolalives in relationship withI.

cembraeonLarix deciduain Europe andC. polonicais associated withI.

typographus,I.amitimusandI.duplicatusliving onPicea abies(Marin et al.

2005). In the present study the same significant differences in growth rate at 32°C

and ecological niche betweenC.laricicolaandC. polonicawere found, too. The

Czech strain ofC.laricicolaformed 29–33 mm large colonies on 2 % malt extract

agar after 8 days at 32 °C, whereas colonies of the Czech strain ofC.polonica

reached only a diameter of 4–10 mm under the same conditions. The strains ofC.

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CZECH MYCOL . 62(1): 59–65, 2010

laricicolaandC.polonicawere obtained fromL.deciduaandP.abies, respec

-

tively. The sequences ofβ-tubulin and MAT-2 HMG box of the Czech strain ofC.

laricicolawere not studied.

The identification of the Czech strain was based on its growth at a temperature

of 32 °C (29–33 mm colonies after 8 days) and specific ecological niche (gallery of

I.cembraefromL.decidua).

A

CKNOWLEDGEMENTS

I would like to thank Petr Šrůtka for helping with collecting material. The pro

-

ject was supported by the Grant Agency of the Czech Republic (project no.

206/05/P279) and by project MZE0002700604 from the Ministry of Agriculture of

the Czech Republic.

R

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