Monitoring diversity patterns of active flagellate taxa in different grassland ecosystems as a tool in terrestrial ecology

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Protist ecology thus far has focused nearly exclusively on functional group models, such as heterotrophic nanoflagellates. Recent investigations, however, indicate high molecular diversity and functional diversification, and habitat specificity of distinct flagellate taxa. The goals of the project are to correlate molecular diversity patterns of selected flagellate taxa (cercomonads, kinetoplastea, chrysomonads) with vegetation type and land use. The overwhelming majority of published DNA-based studies targets the potential diversity of protists since non-active, encysted cells are analyzed with this approach as well. Contrary to this, we will apply and evaluate adequate and specific tools to identify active members of these protistan taxa (i.e. the realized activity). Therefore, we will focus on 18S rRNA-based approaches as a measure to target potentially active flagellate organisms. Ribosomal RNA targeting primers will be used to specifically quantify with quantitative PCR the rRNA of selected flagellate groups in soil samples. The underlying diversity will be investigated on a broader level with a fingerprinting method (T-RFLP), whereas more detailed information shall be obtained by establishing clone libraries. In addition, we aim to develop and apply a rapid quantification tool for soil protist cells combining multiparametric flow cytometry with specific phylogenetic staining. In sum, we aim in particular to establish diversity patterns of ubiquitous active soil flagellates as a monitoring tool for landscape ecology.