Hamburg, Universiẗat, Diss., 1994.

Gene and genome duplications are major mechanisms of eukaryotic genome evolution. Three rounds of genome duplication have occurred in the vertebrate lineage, two rounds (1R, 2R) during early vertebrate evolution and a third round, the fish-specific genome duplication (FSGD), in ray-finned fishes at the base of the teleost lineage. Whole genome duplications (WGDs) are considered to facilitate speciation processes and to provide the genetic raw material for major evolutionary transitions and increases in morphological complexity. In the present study, I have used comparative genomic approaches combining molecular phylogenetic reconstructions, synteny analyses as well as gene function studies (expression analyses and knockdown experiments) to investigate the evolutionary consequences and significance of the three vertebrate WGDs. First, the evolutionary history of the endothelin signaling system consisting of endothelin ligands and receptors was reconstructed. The endothelin system is a key component for the development of a major vertebrate innovation, the neural crest. This analysis shows that the endothelin system emerged in an ancestor of the vertebrate lineage and that its members in extant vertebrate genomes are derived from the vertebrate WGDs. Each round of WGD was followed by co-evolution of the expanding endothelin ligand and receptor repertoires. This supports the importance of genome duplications for the origin and diversification of the neural crest, but also underlines a major role for the co-option of new genes into the neural crest regulatory network. Next, I have studied the impact of the FSGD on the evolution of teleost pigment cell development and differentiation. The investigation of 128 genes showed that pigmentation genes have been preferentially retained in duplicate after the FSGD so that extant teleost genomes contain around 30% more putative pigmentation genes than tetrapods. Large parts of pigment cell regulatory pathways are present in duplicate being potentially involved in teleost pigmentary innovations. There are also important differences in the retention of duplicated pigmentation genes among divergent teleost lineages. Functional studies of pigment synthesis enzymes in zebrafish and medaka, particularly of the tyrosinase family, revealed lineage-specific functional evolution of duplicated pigmentation genes in teleosts, but also pointed to anciently conserved gene functions in vertebrates. These results suggest that the FSGD has facilitated the evolution of the teleost pigmentary system, which is the most complex and diverse among vertebrates. In conclusion, the present study supports a major role of WGDs for phenotypic evolution and biodiversity in vertebrates, particularly in fish. / Gen- und Genomverdopplungen sind wichtige Mechanismen der Genomevolution in Eukaryonten. Im Verlauf der Evolution der Wirbeltiere gab es drei wichtige Genomduplikationen. Zwei Genomverdopplungen (1R, 2R) fanden während der sehr frühen Vertebratenevolution statt. In der Linie der Fische kam es an der Basis der Teleostier zu einer weiteren, fischspezifischen Genomduplikation (FSGD). Man nimmt an, dass Genomduplizierungen Artbildungsprozesse begünstigen und dass sie zusätzliches genetisches Material für wichtige evolutionäre Übergänge und für die Steigerung morphologischer Komplexität erzeugen. In der vorliegenden Arbeit wurden Methoden der vergleichenden und funktionellen Genomik gewählt, um die Auswirkungen und die Bedeutung der drei Genomverdopplungen bei Vertebraten zu untersuchen. Dazu wurden molekularphylogenetische Stammbaumanalysen und Synteniedaten mit Genexpressionsstudien und Knockdown-Experimenten kombiniert. Zunächst wurde die Evolution des Endothelin-Signalsystems rekonstruiert. Dieses besteht aus Endothelin-Liganden und -Rezeptoren und hat eine Schlüsselrolle in die Entwicklung der Neuralleiste. Die Neuralleiste und die von ihr abgeleiteten Zelltypen sind wirbeltierspezifische Innovationen. Die Analyse zeigt, dass das Endothelin-System in einem gemeinsamen Vorfahren der Vertebraten entstanden ist. Die in den Genomen rezenter Vertebraten vorkommenden Komponenten des Endothelin-Systems sind durch die drei Genomverdoppelungen entstanden. Nach jeder der Duplizierungen kam es zur Ko-Evolution der Liganden- und Rezeptorenfamilien. Die Evolution des Endothelin-System unterstreicht daher die Bedeutung der Genomduplizierungen für den Ursprung und die Diversifizierung der Neuralleiste. Sie weist aber auch auf eine wichtige Rolle für die Integrierung neuer Gene in das regulatorische Netzwerk der Neuralleiste hin. Im Weiteren wurde der Einfluss der FSGD auf die Evolution der Pigmentzellentwicklung und differenzierung in Teleostiern untersucht. Die evolutionäre Analyse von 128 Genen zeigte, dass Pigmentierungsgene nach der FSGD bevorzugt in zwei Kopien erhalten geblieben sind. Daher besitzen rezente Teleostier im Vergleich zu Landwirbeltieren zusätzlich ca. 30% mehr Gene mit potentiellen Funktionen für die Pigmentierung. Große Teile der regulatorischen Signalwege in den Pigmentzellen liegen daher als zwei Kopien vor. Diese waren möglicherweise an der Evolution von Innovationen in der Körperfärbung von Teleostiern beteiligt. In der vorliegenden Arbeit wurden auch wichtige Unterschiede zwischen verschiedenen Fischgruppen im Erhalt duplizierter Pigmentierungsgene gefunden. Funktionelle Studien bei Zebrafish und bei Medaka an Enzymen der Pigmentsynthese, insbesondere der Tyrosinase-Familie, gaben Hinweise darauf, dass die funktionelle Evolution duplizierter Pigmentierungsgene in Fischen linienspezifisch verlaufen kann. Die Studien ergaben außerdem, dass bestimmte Funktionen der Pigmentsyntheseenzyme innerhalb der Vertebraten konserviert sind. Die Evolution des Pigmentierungssystems der Fische, welches das vielfältigste und komplexeste innerhalb der Wirbeltiere ist, wurde somit maßgeblich durch die FSGD beeinflusst. Zusammenfassend weisen die Ergebnisse der vorliegenden Arbeit darauf hin, dass die Verdopplung ganzer Genome ein wichtiger Mechanismus der phänotypische Evolution bei Vertebraten ist und damit in besonderem Maße zur ihrer Biodiversität beiträgt.

Konstanz, Univ., Diplomarb., 2001.

Universiẗat, Diss., 2003--Münster (Westfalen).

It is a widely accepted assumption that fish predation controls structure and functioning of aquatic food webs. In the past, however, a large part of effort was concentrated on lakes and reservoirs. Thus, the knowledge about stream ecosystems is much more rudimentary than that for lakes in this respect. The aim of this thesis, therefore, was to describe and assess the effects of fish predation in natural stream ecosystems. For that purpose a reach scale field experiment was set up using an experimental stretch with benthivorous fish and a fishless reference stretch. A wide range of effects of the fish predators on their stream invertebrates prey was studied. To discriminate between lethal and sublethal predation effects, measuring the physiological status of the organisms seemed promising. However, before it was possible to decide whether or not environmental stress, such as predation, might affect the physiological status, the internal control as well as the seasonal and species-specific variability of the energy amount stored had to be assessed. Thus, the concentration and seasonal dynamics of the major energy storage components triglycerides and glycogen were measured in two species of mayflies (Rhithrogena semicolorata and Ephemera danica) with contrasting life cycle strategies. E. danica is a burrowing, semivoltine collector-gatherer, R. semicolorata is univoltine and scrapes periphyton from stones. Although triglycerides are the major energy reserve in both species throughout the whole larval development (> 84 % of total energy storage) their seasonal dynamic differed considerably. In R. semicolorata the triglyceride concentration declined during the last weeks prior to emergence in both sexes. The same pattern was found in female larvae of E. danica, but not in male E. danica. It is suggested that females use triglycerides in the last larval stages for egg maturation, which is completed in the last larval instar. In male E. danica the triglyceride concentrations remained high until emergence, presumably due to their high energy demands as adults for their swarming flights and mating. The difference in seasonal variation of triglycerides between E. danica and R. semicolorata shows the influence of environmental factors on the dynamics of storage components. E. danica lived in a very stable environment (within the substratum). Therefore the dynamic of energy storage components was optimised with respect to maximal reproduction. R. semicolorata on the other hand, suffered from hostile environmental factors such as predation or food limitation due to low periphyton biomass after leaf sprout and following light limitation in spring. Consequently, the concentration of storage components decreased during spring. One conclusion from this study was that the measurement of storage components might reveal sublethal predation effects. However, season and sex of the organisms are important factors as well and have to be considered in the sampling design. To analyse sublethal predation effects behavioural changes due to the presence of benthivorous fish were measured. Drift as a low-energy cost means of migration may enable stream invertebrates to leave risky habitats or may even be a direct escape reaction after a predator encounter. While the control of drift activity by predators has received considerable interest from many researchers, it remains still unclear whether predators reduce or increase drift activity. Drift activity of stream invertebrates was influenced significantly by the presence or absence the two benthivorous fish species gudgeon (Gobio gobio) and stone loach (Barbatula barbatula). Contrary to previous studies gudgeon and stone loach reduced invertebrate drift density and drift activity of Baetis rhodani rather than inducing higher night-time drift. Further, species composition of the invertebrate drift differed significantly between the two stretches. A further conclusion from this study is therefore that drift is not generally a mechanism of active escape from benthos-feeding fish, as previously assumed. In addition, the reduced drift activity in the fish stretch might result in a compensation of the consumptive losses due to fish predation. Thus, in this study design the effects of fish predation on invertebrate community might be underestimated. To detect predation effects on the food web structure the reactions of the grazing mayfly Rhithrogena semicolorata and the shredding amphipod Gammarus pulex to strong predation by benthivorous fish were compared. It has been hypothesised that shredders are generally less vulnerable to fish predation and therefore less likely to be predation-controlled than grazers, because the latter are visible to the predators during their feeding on stone surfaces, while shredders may hide between leaves during foraging. Biomass of G. pulex was significantly reduced in the fish stretch while that of R. semicolorata was not. Since approximately 91 % of the annual production of G. pulex but only 12 % of R. semicolorata production was consumed by benthivorous gudgeon, the observed difference of G. pulex biomass between the fish and reference reach is likely due to a lethal predation effect. However, no sublethal predation effects such as reduced concentration of storage components (triglycerides, glycogen) or reduced reproductive success were observed for both species. Hence, in contrast to the initial hypothesis, in the studied stream the shredder was top-down-controlled, while the grazer was not. It is concluded that top-down control depends on the ecological characteristics of a specific predator-prey pair rather than on trophic guild of the prey. To assess the predation effects on the life history of merolimnic insects and its consequences on fecundity the larval development and emergence of R. semicolorata was studied. It was possible to show lethal and sublethal effects of predation by benthivorous fish (Gobio gobio, Barbatula barbatula). Predation consequently resulted in changes of larval development and population fitness. The presence of two benthivorous fish species (gudgeon and stone loach) led to slower larval development and a delayed emergence. However, no differences in the adult size and fecundity between the fish reach and the reference were observed. Nevertheless, the longer time spent in the larval phase resulted in a higher mortality and therefore in a lower mean population fitness. The presence of gudgeon alone, however, did not seem to influence larval development, growth or time of emergence and consequently fecundity. Further, strong lethal impact of gudgeon could not be detected. Thus, the population fitness measured as the product of adult density and egg number was not reduced by gudgeon alone. It is assumed that the stronger lethal impact in the combined fish experiment is caused mainly by stone loach because the proportion of mayfly consumption by stone loach to mayfly production shortly before emergence was higher than the proportion related to gudgeon. Thus another conclusion is that 1) the impact of predation seems to differ for the fish species and 2) lethal effects have a stronger impact on the population survival than life history changes. Combining the results mentioned above leads to the assumption that predation by benthivorous fish has the potential to shape invertebrate communities and food webs in streams. It was possible to show reductions of benthic densities and mean population fitness. The strength of trophic interactions seemed to be specific for the single predator-prey pairs here. Finally, it can be stated that contrary to previous assumptions consumption of the fish predators seemed to be more important for the prey populations than sublethal predation effects.

Konstanz, Univ., Diss., 2006.

Diss. (sammanfattning) Stockholm : Stockholms universitet, 2008. / Härtill 5 uppsatser.

University, Diss., 2002--Bremen.

Techn. University, Diss., 2003--Berlin.

It is a widely accepted assumption that fish predation controls structure and functioning of aquatic food webs. In the past, however, a large part of effort was concentrated on lakes and reservoirs. Thus, the knowledge about stream ecosystems is much more rudimentary than that for lakes in this respect. The aim of this thesis, therefore, was to describe and assess the effects of fish predation in natural stream ecosystems. For that purpose a reach scale field experiment was set up using an experimental stretch with benthivorous fish and a fishless reference stretch. A wide range of effects of the fish predators on their stream invertebrates prey was studied. To discriminate between lethal and sublethal predation effects, measuring the physiological status of the organisms seemed promising. However, before it was possible to decide whether or not environmental stress, such as predation, might affect the physiological status, the internal control as well as the seasonal and species-specific variability of the energy amount stored had to be assessed. Thus, the concentration and seasonal dynamics of the major energy storage components triglycerides and glycogen were measured in two species of mayflies (Rhithrogena semicolorata and Ephemera danica) with contrasting life cycle strategies. E. danica is a burrowing, semivoltine collector-gatherer, R. semicolorata is univoltine and scrapes periphyton from stones. Although triglycerides are the major energy reserve in both species throughout the whole larval development (> 84 % of total energy storage) their seasonal dynamic differed considerably. In R. semicolorata the triglyceride concentration declined during the last weeks prior to emergence in both sexes. The same pattern was found in female larvae of E. danica, but not in male E. danica. It is suggested that females use triglycerides in the last larval stages for egg maturation, which is completed in the last larval instar. In male E. danica the triglyceride concentrations remained high until emergence, presumably due to their high energy demands as adults for their swarming flights and mating. The difference in seasonal variation of triglycerides between E. danica and R. semicolorata shows the influence of environmental factors on the dynamics of storage components. E. danica lived in a very stable environment (within the substratum). Therefore the dynamic of energy storage components was optimised with respect to maximal reproduction. R. semicolorata on the other hand, suffered from hostile environmental factors such as predation or food limitation due to low periphyton biomass after leaf sprout and following light limitation in spring. Consequently, the concentration of storage components decreased during spring. One conclusion from this study was that the measurement of storage components might reveal sublethal predation effects. However, season and sex of the organisms are important factors as well and have to be considered in the sampling design. To analyse sublethal predation effects behavioural changes due to the presence of benthivorous fish were measured. Drift as a low-energy cost means of migration may enable stream invertebrates to leave risky habitats or may even be a direct escape reaction after a predator encounter. While the control of drift activity by predators has received considerable interest from many researchers, it remains still unclear whether predators reduce or increase drift activity. Drift activity of stream invertebrates was influenced significantly by the presence or absence the two benthivorous fish species gudgeon (Gobio gobio) and stone loach (Barbatula barbatula). Contrary to previous studies gudgeon and stone loach reduced invertebrate drift density and drift activity of Baetis rhodani rather than inducing higher night-time drift. Further, species composition of the invertebrate drift differed significantly between the two stretches. A further conclusion from this study is therefore that drift is not generally a mechanism of active escape from benthos-feeding fish, as previously assumed. In addition, the reduced drift activity in the fish stretch might result in a compensation of the consumptive losses due to fish predation. Thus, in this study design the effects of fish predation on invertebrate community might be underestimated. To detect predation effects on the food web structure the reactions of the grazing mayfly Rhithrogena semicolorata and the shredding amphipod Gammarus pulex to strong predation by benthivorous fish were compared. It has been hypothesised that shredders are generally less vulnerable to fish predation and therefore less likely to be predation-controlled than grazers, because the latter are visible to the predators during their feeding on stone surfaces, while shredders may hide between leaves during foraging. Biomass of G. pulex was significantly reduced in the fish stretch while that of R. semicolorata was not. Since approximately 91 % of the annual production of G. pulex but only 12 % of R. semicolorata production was consumed by benthivorous gudgeon, the observed difference of G. pulex biomass between the fish and reference reach is likely due to a lethal predation effect. However, no sublethal predation effects such as reduced concentration of storage components (triglycerides, glycogen) or reduced reproductive success were observed for both species. Hence, in contrast to the initial hypothesis, in the studied stream the shredder was top-down-controlled, while the grazer was not. It is concluded that top-down control depends on the ecological characteristics of a specific predator-prey pair rather than on trophic guild of the prey. To assess the predation effects on the life history of merolimnic insects and its consequences on fecundity the larval development and emergence of R. semicolorata was studied. It was possible to show lethal and sublethal effects of predation by benthivorous fish (Gobio gobio, Barbatula barbatula). Predation consequently resulted in changes of larval development and population fitness. The presence of two benthivorous fish species (gudgeon and stone loach) led to slower larval development and a delayed emergence. However, no differences in the adult size and fecundity between the fish reach and the reference were observed. Nevertheless, the longer time spent in the larval phase resulted in a higher mortality and therefore in a lower mean population fitness. The presence of gudgeon alone, however, did not seem to influence larval development, growth or time of emergence and consequently fecundity. Further, strong lethal impact of gudgeon could not be detected. Thus, the population fitness measured as the product of adult density and egg number was not reduced by gudgeon alone. It is assumed that the stronger lethal impact in the combined fish experiment is caused mainly by stone loach because the proportion of mayfly consumption by stone loach to mayfly production shortly before emergence was higher than the proportion related to gudgeon. Thus another conclusion is that 1) the impact of predation seems to differ for the fish species and 2) lethal effects have a stronger impact on the population survival than life history changes. Combining the results mentioned above leads to the assumption that predation by benthivorous fish has the potential to shape invertebrate communities and food webs in streams. It was possible to show reductions of benthic densities and mean population fitness. The strength of trophic interactions seemed to be specific for the single predator-prey pairs here. Finally, it can be stated that contrary to previous assumptions consumption of the fish predators seemed to be more important for the prey populations than sublethal predation effects.