TY - GEN A1 - Kunstmann, Ruth Sonja A1 - Scheidt, Tom A1 - Buchwald, Saskia A1 - Helm, Alexandra A1 - Mulard, Laurence A. A1 - Fruth, Angelika A1 - Barbirz, Stefanie T1 - Bacteriophage Sf6 Tailspike Protein for Detection of Shigella flexneri Pathogens T2 - Viruses N2 - Bacteriophage research is gaining more importance due to increasing antibiotic resistance. However, for treatment with bacteriophages, diagnostics have to be improved. Bacteriophages carry adhesion proteins, which bind to the bacterial cell surface, for example tailspike proteins (TSP) for specific recognition of bacterial O-antigen polysaccharide. TSP are highly stable proteins and thus might be suitable components for the integration into diagnostic tools. We used the TSP of bacteriophage Sf6 to establish two applications for detecting Shigella flexneri (S. flexneri), a highly contagious pathogen causing dysentery. We found that Sf6TSP not only bound O-antigen of S. flexneri serotype Y, but also the glucosylated O-antigen of serotype 2a. Moreover, mass spectrometry glycan analyses showed that Sf6TSP tolerated various O-acetyl modifications on these O-antigens. We established a microtiter plate-based ELISA like tailspike adsorption assay (ELITA) using a Strep-tag®II modified Sf6TSP. As sensitive screening alternative we produced a fluorescently labeled Sf6TSP via coupling to an environment sensitive dye. Binding of this probe to the S. flexneri O-antigen Y elicited a fluorescence intensity increase of 80% with an emission maximum in the visible light range. The Sf6TSP probes thus offer a promising route to a highly specific and sensitive bacteriophage TSP-based Shigella detection system. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 472 KW - Shigella flexneri KW - bacteriophage KW - tailspike proteins KW - O-antigen KW - serotyping KW - microtiter plate assay KW - fluorescence sensor Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417831 ER - TY - JOUR A1 - Kunstmann, Ruth Sonja A1 - Scheidt, Tom A1 - Buchwald, Saskia A1 - Helm, Alexandra A1 - Mulard, Laurence A. A1 - Fruth, Angelika A1 - Barbirz, Stefanie T1 - Bacteriophage Sf6 Tailspike Protein for Detection of Shigella flexneri Pathogens JF - Viruses N2 - Bacteriophage research is gaining more importance due to increasing antibiotic resistance. However, for treatment with bacteriophages, diagnostics have to be improved. Bacteriophages carry adhesion proteins, which bind to the bacterial cell surface, for example tailspike proteins (TSP) for specific recognition of bacterial O-antigen polysaccharide. TSP are highly stable proteins and thus might be suitable components for the integration into diagnostic tools. We used the TSP of bacteriophage Sf6 to establish two applications for detecting Shigella flexneri (S. flexneri), a highly contagious pathogen causing dysentery. We found that Sf6TSP not only bound O-antigen of S. flexneri serotype Y, but also the glucosylated O-antigen of serotype 2a. Moreover, mass spectrometry glycan analyses showed that Sf6TSP tolerated various O-acetyl modifications on these O-antigens. We established a microtiter plate-based ELISA like tailspike adsorption assay (ELITA) using a Strep-tag®II modified Sf6TSP. As sensitive screening alternative we produced a fluorescently labeled Sf6TSP via coupling to an environment sensitive dye. Binding of this probe to the S. flexneri O-antigen Y elicited a fluorescence intensity increase of 80% with an emission maximum in the visible light range. The Sf6TSP probes thus offer a promising route to a highly specific and sensitive bacteriophage TSP-based Shigella detection system. KW - Shigella flexneri KW - bacteriophage KW - tailspike proteins KW - O-antigen KW - serotyping KW - microtiter plate assay KW - fluorescence sensor Y1 - 2018 U6 - https://doi.org/10.3390/v10080431 SN - 1999-4915 VL - 10 IS - 8 SP - 1 EP - 18 PB - Molecular Diversity Preservation International (MDPI) CY - Basel ER - TY - GEN A1 - Nagel, Rebecca A1 - Kirschbaum, Frank A1 - Hofmann, Volker A1 - Engelmann, Jacob A1 - Tiedemann, Ralph T1 - Electric pulse characteristics can enable species recognition in African weakly electric fish species T2 - Scientific Reports N2 - Communication is key to a wide variety of animal behaviours and multiple modalities are often involved in this exchange of information from sender to receiver. The communication of African weakly electric fish, however, is thought to be predominantly unimodal and is mediated by their electric sense, in which species-specific electric organ discharges (EODs) are generated in a context-dependent and thus variable sequence of pulse intervals (SPI). While the primary function of the electric sense is considered to be electrolocation, both of its components likely carry information regarding identity of the sender. However, a clear understanding of their contribution to species recognition is incomplete. We therefore analysed these two electrocommunication components (EOD waveform and SPI statistics) in two sympatric mormyrid Campylomormyrus species. In a set of five playback conditions, we further investigated which components may drive interspecific recognition and discrimination. While we found that both electrocommunication components are species-specific, the cues necessary for species recognition differ between the two species studied. While the EOD waveform and SPI were both necessary and sufficient for species recognition in C. compressirostris males, C. tamandua males apparently utilize other, non-electric modalities. Mapped onto a recent phylogeny, our results suggest that discrimination by electric cues alone may be an apomorphic trait evolved during a recent radiation in this taxon. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 470 KW - Dwelling Atlantic Mollies KW - Organ Discharge Patterns KW - Mormyrid Fish KW - Pollimyrus-Isidori KW - Acoustic-Signals KW - Sexual Selection KW - Phylogenetic-Relationships KW - Hypopomus-Occidentalis KW - Convergent Evolution KW - Poecili-Mexicana Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417577 ER - TY - JOUR A1 - Nagel, Rebecca A1 - Kirschbaum, Frank A1 - Hofmann, Volker A1 - Engelmann, Jacob A1 - Tiedemann, Ralph T1 - Electric pulse characteristics can enable species recognition in African weakly electric fish species JF - Scientific Reports N2 - Communication is key to a wide variety of animal behaviours and multiple modalities are often involved in this exchange of information from sender to receiver. The communication of African weakly electric fish, however, is thought to be predominantly unimodal and is mediated by their electric sense, in which species-specific electric organ discharges (EODs) are generated in a context-dependent and thus variable sequence of pulse intervals (SPI). While the primary function of the electric sense is considered to be electrolocation, both of its components likely carry information regarding identity of the sender. However, a clear understanding of their contribution to species recognition is incomplete. We therefore analysed these two electrocommunication components (EOD waveform and SPI statistics) in two sympatric mormyrid Campylomormyrus species. In a set of five playback conditions, we further investigated which components may drive interspecific recognition and discrimination. While we found that both electrocommunication components are species-specific, the cues necessary for species recognition differ between the two species studied. While the EOD waveform and SPI were both necessary and sufficient for species recognition in C. compressirostris males, C. tamandua males apparently utilize other, non-electric modalities. Mapped onto a recent phylogeny, our results suggest that discrimination by electric cues alone may be an apomorphic trait evolved during a recent radiation in this taxon. KW - Dwelling Atlantic Mollies KW - Organ Discharge Patterns KW - Mormyrid Fish KW - Pollimyrus-Isidori KW - Acoustic-Signals KW - Sexual Selection KW - Phylogenetic-Relationships KW - Hypopomus-Occidentalis KW - Convergent Evolution KW - Poecili-Mexicana Y1 - 2018 U6 - https://doi.org/10.1038/s41598-018-29132-z SN - 2045-2322 VL - 8 SP - 1 EP - 12 PB - Nature Publishing Group CY - London ER - TY - GEN A1 - Broeker, Nina K. A1 - Kiele, Franziska A1 - Casjens, Sherwood R. A1 - Gilcrease, Eddie B. A1 - Thalhammer, Anja A1 - Koetz, Joachim T1 - In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620 T2 - Viruses N2 - Gram-negative bacteria protect themselves with an outermost layer containing lipopolysaccharide (LPS). O-antigen-specific bacteriophages use tailspike proteins (TSP) to recognize and cleave the O-polysaccharide part of LPS. However, O-antigen composition and structure can be highly variable depending on the environmental conditions. It is important to understand how these changes may influence the early steps of the bacteriophage infection cycle because they can be linked to changes in host range or the occurrence of phage resistance. In this work, we have analyzed how LPS preparations in vitro trigger particle opening and DNA ejection from the E. coli podovirus HK620. Fluorescence-based monitoring of DNA release showed that HK620 phage particles in vitro ejected their genome at velocities comparable to those found for other podoviruses. Moreover, we found that HK620 irreversibly adsorbed to the LPS receptor via its TSP at restrictive low temperatures, without opening the particle but could eject its DNA at permissive temperatures. DNA ejection was solely stimulated by LPS, however, the composition of the O-antigen dictated whether the LPS receptor could start the DNA release from E. coli phage HK620 in vitro. This finding can be significant when optimizing bacteriophage mixtures for therapy, where in natural environments O-antigen structures may rapidly change. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 469 KW - O-antigen specific phage KW - podovirus KW - HK620 KW - lipopolysaccharide KW - in vitro particle opening KW - tailspike protein Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417493 ER - TY - JOUR A1 - Broeker, Nina K. A1 - Kiele, Franziska A1 - Casjens, Sherwood R. A1 - Gilcrease, Eddie B. A1 - Thalhammer, Anja A1 - Koetz, Joachim T1 - In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620 JF - Viruses N2 - Gram-negative bacteria protect themselves with an outermost layer containing lipopolysaccharide (LPS). O-antigen-specific bacteriophages use tailspike proteins (TSP) to recognize and cleave the O-polysaccharide part of LPS. However, O-antigen composition and structure can be highly variable depending on the environmental conditions. It is important to understand how these changes may influence the early steps of the bacteriophage infection cycle because they can be linked to changes in host range or the occurrence of phage resistance. In this work, we have analyzed how LPS preparations in vitro trigger particle opening and DNA ejection from the E. coli podovirus HK620. Fluorescence-based monitoring of DNA release showed that HK620 phage particles in vitro ejected their genome at velocities comparable to those found for other podoviruses. Moreover, we found that HK620 irreversibly adsorbed to the LPS receptor via its TSP at restrictive low temperatures, without opening the particle but could eject its DNA at permissive temperatures. DNA ejection was solely stimulated by LPS, however, the composition of the O-antigen dictated whether the LPS receptor could start the DNA release from E. coli phage HK620 in vitro. This finding can be significant when optimizing bacteriophage mixtures for therapy, where in natural environments O-antigen structures may rapidly change. KW - O-antigen specific phage KW - podovirus KW - HK620 KW - lipopolysaccharide KW - in vitro particle opening KW - tailspike protein Y1 - 2018 U6 - https://doi.org/10.3390/v10060289 SN - 1999-4915 VL - 10 IS - 6 SP - 1 EP - 15 PB - Molecular Diversity Preservation International (MDPI) CY - Basel ER - TY - GEN A1 - Scherer, Ulrike A1 - Tiedemann, Ralph A1 - Schlupp, Ingo T1 - Male size, not female preferences influence female reproductive success in a poeciliid fish (Poecilia latipinna) BT - a combined behavioural/genetic approach T2 - BMC Research Notes N2 - Objective We investigated the potential role of indirect benefits for female mate preferences in a highly promiscuous species of live-bearing fishes, the sailfin molly Poecilia latipinna using an integrative approach that combines methods from animal behavior, life-history evolution, and genetics. Males of this species solely contribute sperm for reproduction, and consequently females do not receive any direct benefits. Despite this, females typically show clear mate preferences. It has been suggested that females can increase their reproductive success through indirect benefits from choosing males of higher quality. Results Although preferences for large body size have been recorded as an honest signal for genetic quality, this particular study resulted in female preference being unaffected by male body size. Nonetheless, larger males did sire more offspring, but with no effect on offspring quality. This study presents a methodical innovation by combining preference testing with life history measurements—such as the determination of the dry weight of fish embryos—and paternity analyses on single fish embryos. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 468 KW - Fitness KW - Life history KW - Mate choice KW - Microsatellite analysis KW - Offspring weight KW - Paternity analysis KW - Sailfin molly KW - Sexual selection Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417471 ER - TY - JOUR A1 - Scherer, Ulrike A1 - Tiedemann, Ralph A1 - Schlupp, Ingo T1 - Male size, not female preferences influence female reproductive success in a poeciliid fish (Poecilia latipinna) BT - a combined behavioural/genetic approach JF - BMC Research Notes N2 - Objective We investigated the potential role of indirect benefits for female mate preferences in a highly promiscuous species of live-bearing fishes, the sailfin molly Poecilia latipinna using an integrative approach that combines methods from animal behavior, life-history evolution, and genetics. Males of this species solely contribute sperm for reproduction, and consequently females do not receive any direct benefits. Despite this, females typically show clear mate preferences. It has been suggested that females can increase their reproductive success through indirect benefits from choosing males of higher quality. Results Although preferences for large body size have been recorded as an honest signal for genetic quality, this particular study resulted in female preference being unaffected by male body size. Nonetheless, larger males did sire more offspring, but with no effect on offspring quality. This study presents a methodical innovation by combining preference testing with life history measurements—such as the determination of the dry weight of fish embryos—and paternity analyses on single fish embryos. KW - Fitness KW - Life history KW - Mate choice KW - Microsatellite analysis KW - Offspring weight KW - Paternity analysis KW - Sailfin molly KW - Sexual selection Y1 - 2018 U6 - https://doi.org/10.1186/s13104-018-3487-2 SN - 1756-0500 VL - 11 IS - 364 SP - 1 EP - 5 PB - Biomed Central CY - London ER - TY - GEN A1 - Ehrlich, Elias A1 - Gaedke, Ursula T1 - Not attackable or not crackable BT - How pre-and post-attack defenses with different competition costs affect prey coexistence and population dynamics T2 - Ecology and Evolution N2 - It is well-known that prey species often face trade-offs between defense against predation and competitiveness, enabling predator-mediated coexistence. However, we lack an understanding of how the large variety of different defense traits with different competition costs affects coexistence and population dynamics. Our study focusses on two general defense mechanisms, that is, pre-attack (e.g., camouflage) and post-attack defenses (e.g., weaponry) that act at different phases of the predator—prey interaction. We consider a food web model with one predator, two prey types and one resource. One prey type is undefended, while the other one is pre-or post-attack defended paying costs either by a higher half-saturation constant for resource uptake or a lower maximum growth rate. We show that post-attack defenses promote prey coexistence and stabilize the population dynamics more strongly than pre-attack defenses by interfering with the predator’s functional response: Because the predator spends time handling “noncrackable” prey, the undefended prey is indirectly facilitated. A high half-saturation constant as defense costs promotes coexistence more and stabilizes the dynamics less than a low maximum growth rate. The former imposes high costs at low resource concentrations but allows for temporally high growth rates at predator-induced resource peaks preventing the extinction of the defended prey. We evaluate the effects of the different defense mechanisms and costs on coexistence under different enrichment levels in order to vary the importance of bottom-up and top-down control of the prey community. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 466 KW - coexistence KW - competition–defense trade‐off KW - defense against predation KW - functional response KW - indirect facilitation KW - predator–prey cycles Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417391 ER - TY - JOUR A1 - Ehrlich, Elias A1 - Gaedke, Ursula T1 - Not attackable or not crackable BT - How pre-and post-attack defenses with different competition costs affect prey coexistence and population dynamics JF - Ecology and Evolution N2 - It is well-known that prey species often face trade-offs between defense against predation and competitiveness, enabling predator-mediated coexistence. However, we lack an understanding of how the large variety of different defense traits with different competition costs affects coexistence and population dynamics. Our study focusses on two general defense mechanisms, that is, pre-attack (e.g., camouflage)and post-attack defenses (e.g., weaponry) that act at different phases of the predator—prey interaction. We consider a food web model with one predator, two prey types and one resource. One prey type is undefended, while the other one is pre-or post-attack defended paying costs either by a higher half-saturation constant for resource uptake or a lower maximum growth rate. We show that post-attack defenses promote prey coexistence and stabilize the population dynamics more strongly than pre-attack defenses by interfering with the predator’s functional response: Because the predator spends time handling “noncrackable” prey, the undefended prey is indirectly facilitated. A high half-saturation constant as defense costs promotes coexistence more and stabilizes the dynamics less than a low maximum growth rate. The former imposes high costs at low resource concentrations but allows for temporally high growth rates at predator-induced resource peaks preventing the extinction of the defended prey. We evaluate the effects of the different defense mechanisms and costs on coexistence under different enrichment levels in order to vary the importance of bottom-up and top-down control of the prey community. KW - coexistence KW - competition–defense trade‐off KW - defense against predation KW - functional response KW - indirect facilitation KW - predator–prey cycles Y1 - 2018 U6 - https://doi.org/10.1002/ece3.4145 SN - 2045-7758 VL - 8 IS - 13 SP - 6625 EP - 6637 PB - Wiley ER - TY - THES A1 - Ma, Xuemin T1 - Characterization of NAC transcription factors involved in leaf senescence and fruit ripening in tomato Y1 - 2018 ER - TY - THES A1 - Schöne, Anne-Christin T1 - Degradation of Aliphatic Polyesters at the Air-Water Interface - Capabilities of the Langmuir Monolayer Technique Y1 - 2018 ER - TY - THES A1 - Schwarzer, Christian T1 - Climate change, adaptive divergence and their effects on species interactions in European bog-plant communities Y1 - 2018 ER - TY - THES A1 - Hilgers, Leon T1 - From innovation to diversification BT - the genetic basis of the molluscan radula and its adaptive diversification in Tylomelania Y1 - 2018 ER - TY - THES A1 - Schwanhold, Nadine T1 - Die Funktion und Spezifität der Molybdän-Cofaktor-bindenden Chaperone für die Formiat-Dehydrogenasen aus Escherichia coli und Rhodobacter capsulatus Y1 - 2018 ER - TY - THES A1 - Scheffler, Christiane T1 - Studies on plasticity within the universal pattern of growth and developmet of children and adolescents N2 - The anatomically modern human Homo sapiens sapiens is distinguished by a high adaptability in physiology, physique and behaviour in short term changing environmental conditions. Since our environmental factors are constantly changing because of anthropogenic influences, the question arises as to how far we have an impact on the human phenotype in the very sensitive growth phase in children and adolescents. Growth and development of all children and adolescents follow a universal and typical pattern. This pattern has evolved as the result of trade-offs in the 6-7 million years of human evolution. This typically human growth pattern differs from that of other long-living social primate species. It can be divided into different biological age stages, with specific biological, cognitive and socio-cultural signs. Phenotypic plasticity is the ability of an organism to react to an internal or external environmental input with a change in the form, state, and movement rate of activity (West-Eberhard 2003). The plasticity becomes visible and measurable particularly when, in addition to the normal variability of the phenotypic characteristics within a population, the manifestation of this plasticity changes within a relatively short time. The focus of the present work is the comparison of age-specific dimensional changes. The basic of the presented studies are more than 75,000 anthropometric data-sets of children and adolescence from 1980 up today and historical data of height available in scientific literature. Due to reduced daily physical activity, today's 6-18 year-olds have lower values of pelvic and elbow breadths. The observed increase in body height can be explained by hierarchies in social networks of human societies, contrary to earlier explanations (influence of nutrition, good living conditions and genetics). A shift towards a more feminine fat distribution pattern in boys and girls is parallel to the increase in chemicals in our environment that can affect the hormone system. Changing environmental conditions can have selective effects over generations so that that genotype becomes increasingly prevalent whose individuals have a higher progeny rate than other individuals in this population. Those then form the phenotype which allows optimum adaptation to the changes of the environmental conditions. Due to the slow patterns of succession and the low progeny rate (Hawkes et al. 1998), fast visible in the phenotype due to changes in the genotype of a population are unlikely to occur in the case of Homo sapiens sapiens within short time. In the data sets on which the presented investigations are based, such changes appear virtually impossible. The study periods cover 5-30 to max.100 years (based on data from the body height from historical data sets). N2 - Der anatomisch moderne Mensch Homo sapiens sapiens zeichnet sich durch eine hohe Anpassungsfähigkeit von Physiologie, Körperbau und Verhalten an sich kurzfristig ändernde Umweltbedingungen aus. Daraus ergibt sich die Frage inwieweit anthropogene Umweltbedingungen die sehr sensible Wachstumsphase von Kindern und Jugendlichen beeinflussen können. Das universelle und für den Menschen typische Wachstums- und Entwicklungsmuster mit unterschiedlichen biologisch, kognitiv und soziokulturell abgrenzbaren Entwicklungsstadien, welches sich in 6-7 Millionen Jahren menschlicher Evolution herausgebildet hat, unterscheidet sich von dem anderer langlebender sozialer Primaten. Phänotypische Plastizität ist die Fähigkeit eines Organismus sich in Form, Zustand, Aktivitätsrate oder Verhalten an unterschiedliche Umweltbedingungen anzupassen (West-Eberhard 2003). Beim Menschen wird diese Plastizität u.a. sichtbar, wenn sich anthropometrisch bestimmbare Merkmale im Vergleich von Populationen in relativ kurzer Zeit ändern. Der Schwerpunkt der vorliegenden Arbeit ist es, altersspezifische Änderung von Körpermaßen (Skelettbreiten, Körperendhöhe und Fettverteilungsmuster) aufeinanderfolgender Populationen in Abhängigkeit von neuen Umweltparametern zu vergleichen. Dem liegen ca. 75 000 anthropo-metrische Datensätzen von Kindern und Jugendlichen seit 1980 bis heute und historische Datensätze aus der Literatur zugrunde. Aufgrund verringerter alltäglicher Bewegung haben heutige 6-18-Jährige geringere Werte der Becken- und der Ellenbogenbreiten. Die beobachtete Zunahme der Körperhöhe lässt sich entgegen früherer Erklärungen (Einfluss von Ernährung, guter Lebensbedingungen und Genetik) durch Hierarchien in sozialen Netzwerken menschlicher Gesellschaften erklären. Eine Verschiebung zu einem eher weiblichen Fettverteilungsmuster bei Jungen und Mädchen findet sich parallel zur Zunahme von Chemikalien in unserer Umwelt, die das Hormonsystem beeinflussen können. Die beschriebene Plastizität des Phänotyps findet im Rahmen des genetisch manifestierten Wachstumsmusters bei Kindern und Jugendlichen statt. Epigenetische Einflüsse können nicht ausgeschlossen werden, sind aber an Körpermaßdaten per se nicht bestimmbar. Die Veränderung der analysierten Körpermaße unterstreicht, dass der Phänotyp des Menschen sich an veränderte Umweltbedingungen sehr plastisch anpassen kann. Wegen der langsamen Generationenfolge und Entwicklung des Menschen sind derartige eigentlich kurzfristige Veränderungen nur über einen Zeitraum von mindestens 5-30 Jahren zu beobachten. KW - plasticity KW - skeletal breadth measurement KW - Fat Patterning KW - secular trend KW - body height Y1 - 2018 ER - TY - THES A1 - Pham, Phuong Anh T1 - The metabolic significance of the NAD+ salvage pathway and the alternative pathway of respiration in Arabidopsis thaliana Y1 - 2018 ER - TY - THES A1 - Westbury, Michael V. T1 - Unraveling evolution through Next Generation Sequencing T1 - Entschlüsselung von Evolution durch Sequenzierung der nächsten Generation N2 - The sequencing of the human genome in the early 2000s led to an increased interest in cheap and fast sequencing technologies. This interest culminated in the advent of next generation sequencing (NGS). A number of different NGS platforms have arisen since then all promising to do the same thing, i.e. produce large amounts of genetic information for relatively low costs compared to more traditional methods such as Sanger sequencing. The capabilities of NGS meant that researchers were no longer bound to species for which a lot of previous work had already been done (e.g. model organisms and humans) enabling a shift in research towards more novel and diverse species of interest. This capability has greatly benefitted many fields within the biological sciences, one of which being the field of evolutionary biology. Researchers have begun to move away from the study of laboratory model organisms to wild, natural populations and species which has greatly expanded our knowledge of evolution. NGS boasts a number of benefits over more traditional sequencing approaches. The main benefit comes from the capability to generate information for drastically more loci for a fraction of the cost. This is hugely beneficial to the study of wild animals as, even when large numbers of individuals are unobtainable, the amount of data produced still allows for accurate, reliable population and species level results from a small selection of individuals. The use of NGS to study species for which little to no previous research has been carried out on and the production of novel evolutionary information and reference datasets for the greater scientific community were the focuses of this thesis. Two studies in this thesis focused on producing novel mitochondrial genomes from shotgun sequencing data through iterative mapping, bypassing the need for a close relative to serve as a reference sequence. These mitochondrial genomes were then used to infer species level relationships through phylogenetic analyses. The first of these studies involved reconstructing a complete mitochondrial genome of the bat eared fox (Otocyon megalotis). Phylogenetic analyses of the mitochondrial genome confidently placed the bat eared fox as sister to the clade consisting of the raccoon dog and true foxes within the canidae family. The next study also involved reconstructing a mitochondrial genome but in this case from the extinct Macrauchenia of South America. As this study utilised ancient DNA, it involved a lot of parameter testing, quality controls and strict thresholds to obtain a near complete mitochondrial genome devoid of contamination known to plague ancient DNA studies. Phylogenetic analyses confidently placed Macrauchenia as sister to all living representatives of Perissodactyla with a divergence time of ~66 million years ago. The third and final study of this thesis involved de novo assemblies of both nuclear and mitochondrial genomes from brown and striped hyena and focussed on demographic, genetic diversity and population genomic analyses within the brown hyena. Previous studies of the brown hyena hinted at very low levels of genomic diversity and, perhaps due to this, were unable to find any notable population structure across its range. By incorporating a large number of genetic loci, in the form of complete nuclear genomes, population structure within the brown hyena was uncovered. On top of this, genomic diversity levels were compared to a number of other species. Results showed the brown hyena to have the lowest genomic diversity out of all species included in the study which was perhaps caused by a continuous and ongoing decline in effective population size that started about one million years ago and dramatically accelerated towards the end of the Pleistocene. The studies within this thesis show the power NGS sequencing has and its utility within evolutionary biology. The most notable capabilities outlined in this thesis involve the study of species for which no reference data is available and in the production of large amounts of data, providing evolutionary answers at the species and population level that data produced using more traditional techniques simply could not. N2 - Die Sequenzierung des ersten menschlichen Genoms Anfang der 2000er Jahre förderte das Interesse an kostengünstigen und gleichzeitig schnelleren Sequenziertechniken. Dieses Interesse erreichte seinen derzeitigen Höhepunkt in der Einführung des sogenannten Next Generation Sequencings (NGS). Seitdem wurden zahlreiche NGS-Plattformen entwickelt, die alle dem gleichen Prinzip folgen, nämlich das Erzeugen großer Mengen genetischer Information zu relativ geringen Preisen verglichen mit herkömmlichen Methoden wie der Sanger-Sequenzierung. Die neue Leistungsfähigkeit von NGS bedeutete, dass Forscher nicht mehr länger an Organismen gebunden waren an denen bereits seit Jahren geforscht wurde (bspw. Modellorganismen oder der Mensch), sondern ermöglichte eine Verschiebung in Richtung neuerer und unterschiedlicher Arten von Interesse. Dieses Potential hat viele Wissenschaftsfelder positiv beeinflusst innerhalb der Biowissenschaften, u.a. das Feld der Evolutionsbiologie. Forscher haben angefangen sich zunehmend von Modellorganismen in Laboratorien wegzubewegen hinzu wildlebenden, natürlich vorkommenden Populationen und Arten, was unser Verständnis von Evolution maßgeblich erweitert hat. NGS hat mehrere Vorteile aufzuweisen gegenüber den herkömmlichen Sequenziermethoden. Der wohl größte Vorteil ist die Gewinnung genetischer Daten für mehrere Genorte (Loci) gleichzeitig zu einem Bruchteil der bisherigen Kosten. Das ist besonders nützlich für die Untersuchung wildlebender Tiere da, selbst wenn nicht ausreichend viele Individuen vorliegen, die gewonnene Menge an Daten genaue und verlässliche Ergebnisse auf Populations- sowie Artebene für eine kleine Auswahl an Individuen liefert. Die Verwendung von NGS zur Untersuchung von Arten, für die bisher wenig oder gar keine vorherigen Forschungsergebnisse vorliegen sowie die Gewinnung neuartiger Informationen im Bereich Evolution ebenso wie die Erstellung eines Referenzdatensatzes, der der wissenschaftlichen Gemeinschaft zur Verfügung gestellt werden kann, waren der Fokus dieser Arbeit. Zwei Studien in dieser Arbeit setzten ihren Fokus in der Gewinnung noch nicht publizierter, mitochondrialer Genome, die mittels iterative mapping erstellt wurden und so das Vorhandensein einer Referenzsequenz eines nahen Verwandten der untersuchten Art unnötig machten. In beiden Fällen wurden Shotgun Sequenzierungsdaten verwendet. Die so gewonnenen mitochondrialen Genome wurden dann genutzt, um innerartliche Verwandtschaftsverhältnisse mit hilfe von phylogenetischen Analysen zu klären. Die erste Studie befasste sich mit der Rekonstruktion des kompletten mitochondrialen Genoms des Löffelhundes (Otocyon megalotis). Die phylogenetische Analyse des mitochondrialen Genoms positionierten den Löffelhund sicher als Schwestergruppe der Klade bestehend aus Marderhund und echten Füchsen innerhalb der Familie Canidae. Die zweite Studie hat sich ebenfalls mit der Rekonstruktion eines mitochondrialen Genoms auseinandergesetzt, diesmal von einer bereits ausgestorbenen Art Südamerikas, dem Macrauchenia. Da diese Studie auf sehr alter DNA (ancient DNA) basiert, schließt sie viele Parametertests, Qualitätskontrollen sowie strenge Filterkriterien ein um ein fast vollständiges mitochondriales Genom erhalten zu können, frei von den für ancient DNA typischen Kontaminationen. Phylogenetische Analysen positionieren Macrauchenia als Schwestergruppe zu allen anderen lebenden Vertretern der Perissodactyla mit einer Abspaltung vor ~66 Millionen Jahren. Die dritte und letzte Studie dieser Arbeit beinhaltet die de novo Konstruktionen von nukleären und mitochondrialen Genomen der Schabracken- und Streifenhyäne mit Fokus auf demographische, genetische Diversität sowie Populationsgenomische Analysen innerhalb der Schabrackenhyänen. Vorausgehende Studien an der Schabrackenhyäne gaben Hinweise für einen geringen Grad an genomischer Diversität und, waren vielleicht deshalb, bisher nicht in der Lage eine nennenswerte Populationsstruktur der Schabrackenhyäne aufzudecken. Zusätzlich wurde die genomische Diversität mit der von einer Reihe anderer Arten verglichen. Die Ergebnisse zeigen, dass die Schabrackenhyäne die niedrigste genomische Diversität aufweist im Vergleich zu den in dieser Studie verwendeten Arten, was vielleicht mit einem kontinuierlichen und fortschreitenden Rückgang der effektiven Populationsgröße dieser Spezies zu erklären ist, der vor ca. einer Million Jahre eingesetzt hat und dramatisch zugenommen hat zum Ende des Pleistozän. Die Studien dieser Arbeit zeigen das Potential von NGS Sequenzierung und ihren Nutzen innerhalb der Evolutionsbiologie. Die nennenswertesten Anwendungen von NGS, die in dieser Arbeit hervorgehoben wurden, sind zum Einen der Nutzen für Organismen bzw. Arten für die es keine verfügbaren Referenzdaten gibt sowie zum Anderen die Gewinnung von großen Datenmengen, die die Grundlage bilden zur Beantwortung evolutionsbiologischer Fragestellungen auf Art- und Populationsebene, was vorhergegangene, traditionelle Methoden bisher nicht leisten konnten. KW - Next generation sequencing KW - Evolution KW - Hyena KW - Evolution KW - Hyäne KW - Sequenzierung der nächsten Generation Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-409981 ER - TY - THES A1 - Al Fadel, Frdoos T1 - Influence of sphingosine 1-phosphate and its receptor modulators on the development of liver fibrosis Y1 - 2018 ER - TY - GEN A1 - Taron, Ulrike H. A1 - Lell, Moritz A1 - Barlow, Axel A1 - Paijmans, Johanna L. A. T1 - Testing of Alignment Parameters for Ancient Samples BT - Evaluating and Optimizing Mapping Parameters for Ancient Samples Using the TAPAS Tool T2 - Genes N2 - High-throughput sequence data retrieved from ancient or other degraded samples has led to unprecedented insights into the evolutionary history of many species, but the analysis of such sequences also poses specific computational challenges. The most commonly used approach involves mapping sequence reads to a reference genome. However, this process becomes increasingly challenging with an elevated genetic distance between target and reference or with the presence of contaminant sequences with high sequence similarity to the target species. The evaluation and testing of mapping efficiency and stringency are thus paramount for the reliable identification and analysis of ancient sequences. In this paper, we present ‘TAPAS’, (Testing of Alignment Parameters for Ancient Samples), a computational tool that enables the systematic testing of mapping tools for ancient data by simulating sequence data reflecting the properties of an ancient dataset and performing test runs using the mapping software and parameter settings of interest. We showcase TAPAS by using it to assess and improve mapping strategy for a degraded sample from a banded linsang (Prionodon linsang), for which no closely related reference is currently available. This enables a 1.8-fold increase of the number of mapped reads without sacrificing mapping specificity. The increase of mapped reads effectively reduces the need for additional sequencing, thus making more economical use of time, resources, and sample material. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 415 KW - ancient DNA KW - short-read mapping KW - palaeogenomics KW - alignment sensitivity / specificity Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-409683 ER - TY - JOUR A1 - Taron, Ulrike H. A1 - Lell, Moritz A1 - Barlow, Axel A1 - Paijmans, Johanna L. A. T1 - Testing of Alignment Parameters for Ancient Samples BT - Evaluating and Optimizing Mapping Parameters for Ancient Samples Using the TAPAS Tool JF - Genes N2 - High-throughput sequence data retrieved from ancient or other degraded samples has led to unprecedented insights into the evolutionary history of many species, but the analysis of such sequences also poses specific computational challenges. The most commonly used approach involves mapping sequence reads to a reference genome. However, this process becomes increasingly challenging with an elevated genetic distance between target and reference or with the presence of contaminant sequences with high sequence similarity to the target species. The evaluation and testing of mapping efficiency and stringency are thus paramount for the reliable identification and analysis of ancient sequences. In this paper, we present ‘TAPAS’, (Testing of Alignment Parameters for Ancient Samples), a computational tool that enables the systematic testing of mapping tools for ancient data by simulating sequence data reflecting the properties of an ancient dataset and performing test runs using the mapping software and parameter settings of interest. We showcase TAPAS by using it to assess and improve mapping strategy for a degraded sample from a banded linsang (Prionodon linsang), for which no closely related reference is currently available. This enables a 1.8-fold increase of the number of mapped reads without sacrificing mapping specificity. The increase of mapped reads effectively reduces the need for additional sequencing, thus making more economical use of time, resources, and sample material. KW - ancient DNA KW - short-read mapping KW - palaeogenomics KW - alignment sensitivity / specificity Y1 - 2018 U6 - https://doi.org/10.3390/genes9030157 SN - 2073-4425 VL - 9 IS - 3 SP - 1 EP - 12 PB - Molecular Diversity Preservation International CY - Basel ER - TY - JOUR A1 - Schedina, Ina Maria A1 - Groth, Detlef A1 - Schlupp, Ingo A1 - Tiedemann, Ralph T1 - The gonadal transcriptome of the unisexual Amazon molly Poecilia formosa in comparison to its sexual ancestors, Poecilia mexicana and Poecilia latipinna JF - BMC Genomics N2 - Background The unisexual Amazon molly (Poecilia formosa) originated from a hybridization between two sexual species, the sailfin molly (Poecilia latipinna) and the Atlantic molly (Poecilia mexicana). The Amazon molly reproduces clonally via sperm-dependent parthenogenesis (gynogenesis), in which the sperm of closely related species triggers embryogenesis of the apomictic oocytes, but typically does not contribute genetic material to the next generation. We compare for the first time the gonadal transcriptome of the Amazon molly to those of both ancestral species, P. mexicana and P. latipinna. Results We sequenced the gonadal transcriptomes of the P. formosa and its parental species P. mexicana and P. latipinna using Illumina RNA-sequencing techniques (paired-end, 100 bp). De novo assembly of about 50 million raw read pairs for each species was performed using Trinity, yielding 106,922 transcripts for P. formosa, 115,175 for P. latipinna, and 133,025 for P. mexicana after eliminating contaminations. On the basis of sequence similarity comparisons to other teleost species and the UniProt databases, functional annotation, and differential expression analysis, we demonstrate the similarity of the transcriptomes among the three species. More than 40% of the transcripts for each species were functionally annotated and about 70% were assigned to orthologous genes of a closely related species. Differential expression analysis between the sexual and unisexual species uncovered 2035 up-regulated and 564 down-regulated genes in P. formosa. This was exemplary validated for six genes by qRT-PCR. Conclusions We identified more than 130 genes related to meiosis and reproduction within the apomictically reproducing P. formosa. Overall expression of these genes seems to be down-regulated in the P. formosa transcriptome compared to both ancestral species (i.e., 106 genes down-regulated, 29 up-regulated). A further 35 meiosis and reproduction related genes were not found in the P. formosa transcriptome, but were only expressed in the sexual species. Our data support the hypothesis of general down-regulation of meiosis-related genes in the apomictic Amazon molly. Furthermore, the obtained dataset and identified gene catalog will serve as a resource for future research on the molecular mechanisms behind the reproductive mode of this unisexual species. KW - Differential gene expression KW - Gynogenesis KW - Hybrid speciation KW - Meiosis KW - Poecilia formosa KW - Poecilia latipinna KW - Poecilia mexicana Y1 - 2018 U6 - https://doi.org/10.1186/s12864-017-4382-2 SN - 1471-2164 VL - 19 IS - 12 SP - 1 EP - 18 PB - BioMed Central CY - London ER - TY - GEN A1 - Schedina, Ina Maria A1 - Groth, Detlef A1 - Schlupp, Ingo A1 - Tiedemann, Ralph T1 - The gonadal transcriptome of the unisexual Amazon molly Poecilia formosa in comparison to its sexual ancestors, Poecilia mexicana and Poecilia latipinna N2 - Abstract Background The unisexual Amazon molly (Poecilia formosa) originated from a hybridization between two sexual species, the sailfin molly (Poecilia latipinna) and the Atlantic molly (Poecilia mexicana). The Amazon molly reproduces clonally via sperm-dependent parthenogenesis (gynogenesis), in which the sperm of closely related species triggers embryogenesis of the apomictic oocytes, but typically does not contribute genetic material to the next generation. We compare for the first time the gonadal transcriptome of the Amazon molly to those of both ancestral species, P. mexicana and P. latipinna. Results We sequenced the gonadal transcriptomes of the P. formosa and its parental species P. mexicana and P. latipinna using Illumina RNA-sequencing techniques (paired-end, 100 bp). De novo assembly of about 50 million raw read pairs for each species was performed using Trinity, yielding 106,922 transcripts for P. formosa, 115,175 for P. latipinna, and 133,025 for P. mexicana after eliminating contaminations. On the basis of sequence similarity comparisons to other teleost species and the UniProt databases, functional annotation, and differential expression analysis, we demonstrate the similarity of the transcriptomes among the three species. More than 40% of the transcripts for each species were functionally annotated and about 70% were assigned to orthologous genes of a closely related species. Differential expression analysis between the sexual and unisexual species uncovered 2035 up-regulated and 564 down-regulated genes in P. formosa. This was exemplary validated for six genes by qRT-PCR. Conclusions We identified more than 130 genes related to meiosis and reproduction within the apomictically reproducing P. formosa. Overall expression of these genes seems to be down-regulated in the P. formosa transcriptome compared to both ancestral species (i.e., 106 genes down-regulated, 29 up-regulated). A further 35 meiosis and reproduction related genes were not found in the P. formosa transcriptome, but were only expressed in the sexual species. Our data support the hypothesis of general down-regulation of meiosis-related genes in the apomictic Amazon molly. Furthermore, the obtained dataset and identified gene catalog will serve as a resource for future research on the molecular mechanisms behind the reproductive mode of this unisexual species. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 412 KW - Differential gene expression KW - Gynogenesis KW - Hybrid speciation KW - Meiosis KW - Poecilia formosa KW - Poecilia latipinna KW - Poecilia mexicana Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-409299 ER - TY - THES A1 - Abd Allah Salem, Mohamed T1 - Comparative and systemic metabolomic analysis of the model plant Arabidopsis thaliana after perturbing the essential Target of Rapamycin (TOR) pathway Y1 - 2018 ER - TY - THES A1 - Robalo, João Ramiro Alavedra Mendes T1 - Investigating the role of fluorinated amino acids on protein structure and function using simulation Y1 - 2018 ER -