Estimation of AM fungal colonization - Comparability and reliability of classical methods.

The characterization of mycorrhizal status in hosts can be a good indicator of symbiotic associations in inoculation experiments or in ecological research. The most common microscopic-based observation methods, such as (i) the gridline intersect method, (ii) the magnified intersections method and (iii) the five-class system of Trouvelot were tested to find the most simple, easily executable, effective and objective ones and their appropriate parameters for characterization of mycorrhizal status. In a pot experiment, white clover (Trifolium repens L.) host plant was inoculated with 6 (BEG144; syn. Rhizophagus intradices) in pumice substrate to monitor the AMF colonization properties during host growth. Eleven (seven classical and four new) colonization parameters were estimated by three researchers in twelve sampling times during plant growth. Variations among methods, observers, parallels, or individual plants were determined and analysed to select the most appropriate parameters and sampling times for monitoring. The comparability of the parameters of the three methods was also tested. As a result of the experiment classical parameters were selected for hyphal colonization: colonization frequency in the first stage or colonization density in the later period, and arbuscular richness of roots. A new parameter was recommended to determine vesicule and spore content of colonized roots at later stages of symbiosis.