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New Zealand Plant Protection 65 (2012): 296

Method for spore production of Leptosphaeria maculans

S. Lob, M.V. Jaspers, H.J. Ridgway and E.E. Jones


To enable the epidemiology of Leptosphaeria maculans, a pathogen of brassicas, to be studied a method for producing spores was required. Sporulation of L. maculans was assessed on V8-juice agar and PDA under two light conditions. Five L. maculans isolates subcultured with three replicates on each agar were incubated under continuous cool white fluorescent lights at room temperature or 12:12 h light:dark at 20C. After 9 days incubation, colony diameter was measured and spores counted. Spore suspensions were produced by pouring 10 ml of sterile water onto each colony and scraping the surface with a sterile glass rod, and spores counted using a haemocytometer. For all isolates colony diameter was greater on V8-juice agar (37.2-52.6 mm) compared to PDA (12.3-50.1 mm) under both conditions. Isolates subcultured on V8-juice agar (9.79106/plate) gave a higher spore concentration than PDA (3.55105/plate). Light condition significantly affected spore production but not colony diameter as the highest spore suspension concentration was obtained from those isolates subcultured on V8- juice agar under continuous cool white fluorescent at room temperature (1.86107/plate) while the lowest concentration was obtained from isolates subcultured on PDA at 20C with 12:12 h light:dark (9.00104/plate). The results have determined the best method for spore production for subsequent experiments.

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