Establishing the twig method for investigations on pollen characteristics of allergenic tree species.

The twig method in climate chambers has been shown to successfully work as a proxy for outdoor manipulations in various experimental setups. This study was conducted to further establish this method for the investigation of allergenic pollen from tree species (hazel, alder, and birch). Direct comparison under outdoor conditions revealed that the cut twigs compared to donor trees were similar in the timing of flowering and the amount of pollen produced. Cut twigs were able to flower in climate chambers and produced a sufficient amount of pollen for subsequent laboratory analysis. The addition of different plant or tissue fertilizers in the irrigation of the twigs did not have any influence; rather, the regular exchange of water and the usage of fungicide were sufficient for reaching the stage of flowering. In the experimental setup, the twigs were cut in different intervals before the actual flowering and were put under warming conditions in the climate chamber. An impact of warming on the timing of flowering/pollen characteristics could be seen for the investigated species. Therefore, the twig method is well applicable for experimental settings in pollen research simulating, e.g., accelerated warming under climate change.