I am doing detached leaf inoculations on potato leaflets with several alternaria species. I am just putting a drop of spore suspension (high concentration, more than 10^5) on the middle of the adaxial surface. No wounding of the leaflet.
The leaflets are sealed in trays with water for high humidity and temperature is 21C with 10 hours of 10 lux light.
Lesions are forming very late, takes about a week. On most leaves they do not form at all.
Any ideas how to optimize the procedure ? I believe wounding of the leaf is important and will use this for next round of experiments.
Hi,
You can try different methods of inoculations like covering the whole leaf with the suspension (with a simple sprayer), thus you can compare the efficiency of these methods.
You use high concentracion of spore suspension (10^5) but maybe you need to apply in even higher rate like 10^7.
Hi: If your isolate is no aggressive you will not get a good infection, you must isolate many isolates of the pathogen and chek the pathogenicity and use the most aggressive in your experiment.
You should consider the virulence of iisolate and host resistance. In addition to this you could also try changing light exposure of the leaves to 16 h and try to do inoculation in the abaxial surface.
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