sdu faculty of forestry journal special edition 2009 - Orman Fakültesi

SDÜ ORMAN FAKÜLTESİ DERGİSİ
Figure 1: Abundant pycnidia of Diplodia pinea on a gall induced by the Cooley spruce
gall adelgid.
2. MATERIAL AND METHODS
Galls were collected in summer from a single mature Colorado blue spruce tree
(approximately 12 m tall) growing as an ornamental on the University of
Wisconsin-Madison campus (43.07 N, 89.40 W). Shoots on this tree did not
exhibit symptoms attributable to D. pinea. Two ornamental Austrian pines (Pinus
nigra Arnold) that were damaged by shoot blight and bore D. pinea pycnidia on
shoots and cones were located within approximately 20 m. The galls that were
collected had matured to release nymphs the previous year and were dead. Ten
arbitrarily selected galls were collected from one branch at each of the four
cardinal directions in the top, middle, and bottom thirds of the tree crown (120
galls total). Galls were bagged separately and frozen until extraction of conidia.
Galls were processed individually using procedures similar to those described
for pine cones by Munck and Stanosz (2009). Each gall was placed in a 100 ml
plastic beaker containing 50 ml of distilled water with 2 drops of Tween 80 l -1
(Fisher Scientific Company, Fair Lawn, NJ) and washed for 3 h on a rotary shaker
at 110 rpm. The resulting suspension was then filtered through 0.8 µm pore size
filters printed with a grid to delineate 3 mm x 3 mm squares (Cat. No.:
AAWG047SP, Millipore Corporation, Billerica, MA). The cup with the gall then
was rinsed with an additional 50 ml of distilled water with Tween and this liquid
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