Temporal temperature gradient gel electrophoresis (TTGE) analysis of arbuscular mycorrhizal fungi associated with selected plants from saline and metal polluted environments

Some plants are more mycorrhizal than others and mycorrhizal colonisation of plants in extreme environments is frequently additionally reduced due to decreased spore density and/or diversity and therefore frequently overlooked. We analysed two plant species from both metal polluted and saline enriched soils with differing mycorrhizal colonisation levels/status using classical and molecular methods. The selected plant species were Sesleria caerulea (L.) Ard. and Thlaspi praecox Wulfen from a metal polluted site, and Limonium angustifolium (Tausch) Degen [Statice serotina Rchb., L. vulgare Mill. subsp. Serotinum (Rchb.) Gams] and Salicornia europaea L. from the Sečovlje salterns in Slovenia. Despite the high mycorrhizal frequencies (F%) observed, the presence of arbuscules (A%) was at best low in S. caerulea and T. praecox, and undetectable in L. angustifolium and S. europaea. Temporal temperature gradient gel electrophoresis (TTGE) was applied to field-collected samples from both burdened environments and proved to be an effective technique for rapid profiling and identification of arbuscular mycorrhizal fungi (AMF). Sequencing and phylogenetic analysis confirmed the association of AMF of the genus Glomus with roots of all four plant species. This is the first report on the identification and profiling of Glomeromycota in the field-collected Cd/Zn metal hyperaccumulator T. praecox growing at a highly metal polluted site, as well as in L. angustifolium and S. europaea collected in a saline environment. The identification of AMF from both ecosystems only partially resembles previous identifications on the basis of spores.