Covering ground: insights into soil ecology by molecular monitoring of nematode assemblages

Soil performs numerous functions, which allow us to produce food and feed and provide us with clean freshwater. These functions rely on the high diversity of organisms residing in soils. Within the high complexity of the soil food web, nematodes, worm-shaped animals belonging to the phylum Nematoda, are an informative group for assessing the status of a soil-dwelling community due to their ubiquity, abundance and trophic diversity. Although nematodes also possess several other assets favourable for a biological indicator of soil ecosystems (e.g. easy extractability, differential sensitivities to disturbances, ecological interpretability), their microscopic identification demands a considerable amount of expertise and time because of their relatively conserved morphology. Hence, the use of a molecular method for the identification and quantification of nematode assemblages has the potential to lift practical limitations and allows for more intensive sampling schemes. The aim of the research described in this thesis was to assess the suitability of molecular taxon-specific assays, developed on the basis of a phylum-wide molecular framework of 2,400 full-length small subunit ribosomal DNA sequences, for the monitoring of nematode assemblages in field experiments. The method was applied to monitor the impact of different types of disturbances on the soil food web, i.e. agricultural practices, invasive plant species and the effects of genetically modified crop (potato). The second chapter of this thesis presents the background of the molecular method and the results of its first field application. It demonstrates the suitability of this method for use in extensive field experiments and the results of this study reveal distinct seasonal fluctuations between nematode genera classified to belong to the same feeding type group. A distinct response of nematode genera within trophic groups – taxonomically diverse groups of nematodes having the nature of their main food source as a common denominator – was also observed in the study described in Chapter 3. In this chapter, a study was conducted to investigate the impact of an invasive plant species called Giant goldenrod (Solidago gigantea, native to North America) on the plant community as well as on different trophic levels of the soil food web. In addition to monitoring the nematode community, pH and fungal biomass were measured in plots invaded or uninvaded by Giant goldenrod in two contrasting habitats. The results revealed that, in addition to outcompeting native plant species, this invader also reduced pH and increased fungal biomass in the soil of both habitats. Based on the results concerning the nematode community, the impact on the soil food web seemed to be selective since the local increase of fungal biomass appeared to benefit only one fungivorous nematode lineage of the three present in the field. This suggests that invasion by Giant goldenrod only stimulates one part of the fungal community. Contrary to the preceding chapters, the effects of different disturbances on the nematode community were studied in an arable setting in chapters 4 and 5. Chapter 4 describes a field experiment in which the impact of biofumigation, a pest control measure, on the nematode community was assessed. Biofumigation is considered as an alternative for the use of synthetic fumigants and entails the incorporation of mulched brassicaceous plant material, which, upon de-compartmentalisation, releases general biocides called isothiocyanates. In our experiment, these compounds as well as their precursors could not be related to the effects observed for the nematode community. Therefore, changes in nematode assemblages are more likely to be related to the intense mechanical disturbance and green manure – the addition of a large quantity of fresh plant material to the topsoil – rather than the release of isothiocyanates from the plant material Chapter 5 presents a field experiment in which the possible belowground side effects of a waxy starch GM potato, a genetically modified plant blocked in its amylose biosynthesis, were investigated. The nematode community was monitored during the growing season of this GM variety, its parental line and four other conventional potato cultivars in two experimental fields. Although we observed clear effects of location and time, no GM-related effects were observed on the nematode community. Our results, in line with previous studies concerning the microbial community, indicate there are no observable, non-transient effects related to this particular GM trait on the soil food web during the growing season. Overall, the results presented in this thesis demonstrate that, first of all, the developed molecular approach is suitable as a tool for the quantitative monitoring of nematode assemblages in field experiments, and, secondly, how a molecular monitoring method based on nematode taxon-specific DNA motifs can be exploited to get new insights into the ecology of terrestrial nematodes and – more in general – into the ecological functioning of this obscure, highly biodiverse and poorly understood habitat below our feet