sdu faculty of forestry journal special edition 2009 - Orman Fakültesi

2. MATERIAL AND METHODS
SDÜ ORMAN FAKÜLTESİ DERGİSİ
Ash twigs (4-8 mm diameter) with dieback (comprising a necrotic distal portion and
a proximal living portion) were collected and transferred to the laboratory. The surface
were disinfected by spraying with concentrated ethanol and superficially dried on a
clean bench. The tissue samples (diameter 2-5 mm, about 2-3 mm long), were
dissected from the sapwood below necrotic lesions, after bark removal. Samples were
then surface sterilized by immersion in sodium hypochlorite (7 - 10 %) for 60 - 90 sec,
then immersed in 96% ethanol for 60 - 90 sec, washed in sterilized water and placed on
the medium. Tissue samples were aseptically transferred on malt extract agar (MEA 3;
30 g/L, peptone 5 g/L, agar 15 g/L) and, according to Kowalski (2006), streptomycin
(100 mg/L) added after autoclaving. Identification was made on the bases of colony
morphology and microscopic features.
3. RESULTS AND DISCUSSION
Ash dieback associated with bark necroses and withering of young shoots was
recorded in several areas in the CR during 2004 – 2008. The locations affected
include mostly all area of the CR, eg. Beskydy Mts., Jeseniky Mts., Giant Mts.,
Bayerischer Wald Mts., Central Bohemia, Prague, Eastern Bohemia, Czech
Moravian Highland, the area at the junction of the Thaya and Morava Rivers, and
along the Czech, Austrian and Slovak borders. Ash dieback has extended across the
entire country since 2004. The symptoms were also noted in nurseries, especially
on saplings in South Moravia. However wooly aphids from genus Prociphilus spp.
were observed in this areas abundantly as a important harmful factor in early
spring.
The first record of Chalara fraxinea in the CR originated from samples
collected at Drahany Highland, Arboretum Krtiny, from Fraxinus excelsior and in
some other locations (Jankovský and Holdenrieder 2009). Ash diaback were
observed on Fraxinus excelsior and its cultivars, especially sensible is F. excelsior
cv. „Pendula“, and F. angustifolia.
On the area of South Moravia samples has been taken especially from the
district of Židlochovice forest enterprise, close to Austrian and Slovak borders.
Health condition of young forest stands is serious on some locations and there is a
danger of large damages of these young plantations. The culture of C. fraxinea
origin from flood-plain forest by the village Tvrdonice in this area. It is located
near of the boarder with Slovakia. The other samples were collected in the Ivaň
area (adjacent to the Nove mlyny reservoir) and Soutok game preserve (National
nature reserve Cahnov).
The occurrence of ash decline is continually monitored, especially in the area of
south Moravia.
According to the experience with symptoms of ash wilting we can assume that
C. fraxinea is spread through the whole area of the CR.
125