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

到目前为止，原生生物生态学几乎完全集中在功能基团模型上，如异养纳米鞭毛虫。然而，最近的研究表明，不同的鞭毛类群具有高度的分子多样性和功能多样性，以及栖息地的特殊性。该项目的目标是将选定的鞭毛类群(尾藻、动藻、金藻)的分子多样性模式与植被类型和土地利用相关联。绝大多数已发表的基于DNA的研究针对的是原生生物的潜在多样性，因为非活跃的包膜细胞也是用这种方法分析的。与此相反，我们将应用和评价适当和具体的工具来确定这些原分类群的活跃成员(即已实现的活动)。因此，我们将重点放在基于18S rRNA的方法上，作为一种针对潜在活性鞭毛生物的措施。核糖体核糖核酸将被用来通过定量聚合酶链式反应对土壤样本中特定鞭毛群的rRNA进行特异性定量。将使用指纹分析方法(T-RFLP)在更广泛的水平上调查潜在的多样性，而将通过建立克隆文库获得更详细的信息。此外，我们的目标是开发和应用一种结合多参数流式细胞术和特定系统发育染色的土壤原生生物细胞快速定量工具。总之，我们的目标是建立无处不在的活跃土壤鞭毛虫的多样性格局，作为景观生态的监测工具。

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.