Institut für Biologie

Browse

Recent Submissions

Now showing 1 - 20 of 161
  • Publication
    tsCRISPR based identification of Rab proteins required for the recycling of Drosophila TRPL ion channel
    (2024) Zeger, Matthias; Stanisławczyk, Lena Sarah; Bulić,Marija; Binder, Andrea Maria; Huber, Armin
    In polarized cells, the precise regulation of protein transport to and from the plasma membrane is crucial to maintain cellular function. Dysregulation of intracellular protein transport in neurons can lead to neurodegenerative diseases such as Retinitis Pigmentosa, Alzheimer’s and Parkinson’s disease. Here we used the light-dependent transport of the TRPL (transient receptor potential-like) ion channel in Drosophila photoreceptor cells to study the role of Rab proteins in TRPL recycling. TRPL is located in the rhabdomeric membrane of dark-adapted flies, but it is transported out of the rhabdomere upon light exposure and localizes at the Endoplasmatic Reticulum within 12 h. Upon subsequent dark adaptation, TRPL is recycled back to the rhabdomeric membrane within 90 min. To screen for Rab proteins involved in TRPL recycling, we established a tissue specific (ts) CRISPR/Cas9-mediated knock- out of individual Rab genes in Drosophila photoreceptors and assessed TRPL localization using an eGFP tagged TRPL protein in the intact eyes of these mutants. We observed severe TRPL recycling defects in the knockouts of Rab3, Rab4, Rab7, Rab32, and RabX2. Using immunohistochemistry, we further showed that Rab3 and RabX2 each play a significant role in TRPL recycling and also influence TRPL transport. We localized Rab3 to the late endosome in Drosophila photoreceptors and observed disruption of TRPL transport to the ER in Rab3 knock-out mutants. TRPL transport from the ER to the rhabdomere ensues from the trans-Golgi where RabX2 is located. We observed accumulated TRPL at the trans-Golgi in RabX2 knock-out mutants. In summary, our study reveals the requirement of specific Rab proteins for different steps of TRPL transport in photoreceptor cells and provides evidence for a unique retrograde recycling pathway of TRPL from the ER via the trans-Golgi
  • Publication
    Metabolic rewiring enables ammonium assimilation via a non‐canonical fumarate‐based pathway
    (2024) Mardoukhi, Mohammad Saba Yousef; Rapp, Johanna; Irisarri, Iker; Gunka, Katrin; Link, Hannes; Marienhagen, Jan; de Vries, Jan; Stülke, Jörg; Commichau, Fabian M.
    Glutamate serves as the major cellular amino group donor. In Bacillus subtilis, glutamate is synthesized by the combined action of the glutamine synthetase and the glutamate synthase (GOGAT). The glutamate dehydrogenases are devoted to glutamate degradation in vivo. To keep the cellular glutamate concentration high, the genes and the encoded enzymes involved in glutamate biosynthesis and degradation need to be tightly regulated depending on the available carbon and nitrogen sources. Serendipitously, we found that the inactivation of the ansR and citG genes encoding the repressor of the ansAB genes and the fumarase, respectively, enables the GOGAT-deficient B. subtilis mutant to synthesize glutamate via a non-canonical fumarate-based ammonium assimilation pathway. We also show that the de-repression of the ansAB genes is sufficient to restore aspartate prototrophy of an aspB aspartate transaminase mutant. Moreover, in the presence of arginine, B. subtilis mutants lacking fumarase activity show a growth defect that can be relieved by aspB overexpression, by reducing arginine uptake and by decreasing the metabolic flux through the TCA cycle.
  • Publication
    Taeniidae in Namibian wildlife with emphasis on lion, cheetah, and African wild dog
    (2024) Aschenborn, Ortwin; Mackenstedt, Ute
    An opportunic survey for Echinococcus spp. in wild mammals was conducted in seven distinct stuy areas throughout Namibia, representing all major ecosystems, between 2012 and 2021. In total, 184 individually attributable faeces and 40 intestines were collected from eight species of carnivores, and 300 carcasses or organs of thirteen species of ungulates were examined for Echinococcus cysts. Nested PCR and sequencing of the mitochondrial nad1 gene led to the identification of five species of the Echinococcus granulosus sensu lato complex. Echinococcus canadensis G6/7 was found throughout Namibia at low frequency in lions, cheetahs, African wild dogs, black-backed jackals and oryx antelopes. Echinococcus equinus was present only in northern Namibia, locally at high frequency in lions, black-backed jackals and plains zebras. Echinococcus felidis was found only in one small area in the north-east of Namibia, but with high frequency in lions and warthogs. Echinococcus granulosus sensu stricto was identified only in two African wild dogs in the north-east of Namibia, and Echinococcus ortleppi occurred in central and southern Namibia in black-backed jackals and oryx antelopes. The development of fertile cysts indicated active intermediate host roles of oryx antelopes for E. canadensis and E. ortleppi, of warthogs for E. felidis, and of plains zebras for E. equinus. Our data support earlier hypotheses of exclusive or predominant wildlife life-cycles for E. felidis involving lions and warthogs, and – in Namibia – for E. equinus involving lions and/or black-backed jackals and plains zebras. Our data further support an interlink of wild and domestic transmission for E. ortleppi. A possible involvement of livestock and domestic dogs in transmission of E. canadensis G6/7 and E. granulosus s.s., the two parasite species with highest zoonotic potential, is uncertain for Namibia and needs further investigation. The present study was conducted in the isolated desert town of Oranjemund in the far south of Namibia. It is an extremely arid region where no livestock husbandry is practiced and only animals adapted to the desert can be found. However, in and around the city, artifi cial irrigation maintains lush green patches of grass that attract wild animals, in particular oryx antelopes (Oryx gazella). In 2015 four oryx antelopes were euthanised due to poor conditions and a post-mortem examination was conducted. Two were found positive for cystic echinococcosis and 16 cysts were collected for molecular analyses. In addition, faecal samples from black-backed jackals (n=5) and domestic dogs (n=9), which were regularly observed to feed on oryx carcasses, were collected and taeniid eggs isolated. Parasite species identifi cation of the cysts and eggs was done by amplifying and se- quencing the mitochondrial nad1 gene. Both oryx antelopes were found infected with E. ortleppi and one co-infected with E. canadensis G6/7. Both Echinococcus species were able to develop fertile cysts in oryx, making oryx antelopes competent hosts for these parasites. Therefore, the analysis of faecal samples was of high interest and although the numbers were quite small, taeniid eggs were found in three out of fi ve faecal samples of jackals and in all nine dog samples. However, species determination was only successful with two jackal and one dog sample. All three were positive for E. canadensis G6/7. The absence of E. ortleppi may be due to the low number of faecal samples examined. In our small study, we discovered a rather unique lifecycle of Echinococcus spp. between jackals and domestic dogs as defi nitive hosts and oryx antelopes as intermediate hosts. Here, the presence of E. canadensis G6/7 is of particular concern, as it is the second most important causative agent of CE in humans.
  • Publication
    The emergence and dynamics of tick-borne Encephalitis Virus in a new endemic region in Southern Germany
    (2022) Lang, Daniel; Chitimia-Dobler, Lidia; Bestehorn-Willmann, Malena; Lindau, Alexander; Drehmann, Marco; Stroppel, Gabriele; Hengge, Helga; Mackenstedt, Ute; Kaier, Klaus; Dobler, Gerhard; Borde, Johannes
    Tick-borne encephalitis (TBE) is the most important viral tick-borne infection in Europe and Asia. It is emerging in new areas. The mechanisms of emergence are fairly unknown or speculative. In the Ravensburg district in southern Germany, TBE emerged, mainly over the last five years. Here, we analyzed the underlying epidemiology in humans. The resulting identified natural foci of the causal TBE virus (TBEV) were genetically characterized. We sampled 13 potential infection sites at these foci and detected TBEV in ticks (Ixodes ricinus) at eight sites. Phylogenetic analysis spurred the introduction of at least four distinct TBEV lineages of the European subtype into the Ravensburg district over the last few years. In two instances, a continuous spread of these virus strains over up to 10 km was observed.
  • Publication
    Central carbon metabolism, sodium-motive electron ransfer, and ammonium formation by the vaginal pathogen Prevotella bivia
    (2021) Schleicher, Lena; Herdan, Sebastian; Fritz, Günter; Trautmann, Andrej; Seifert, Jana; Steuber, Julia
    Replacement of the Lactobacillus dominated vaginal microbiome by a mixed bacterial population including Prevotella bivia is associated with bacterial vaginosis (BV). To understand the impact of P. bivia on this microbiome, its growth requirements and mode of energy production were studied. Anoxic growth with glucose depended on CO2 and resulted in succinate formation, indicating phosphoenolpyruvate carboxylation and fumarate reduction as critical steps. The reductive branch of fermentation relied on two highly active, membrane-bound enzymes, namely the quinol:fumarate reductase (QFR) and Na+-translocating NADH:quinone oxidoreductase (NQR). Both enzymes were characterized by activity measurements, in-gel fluorography, and VIS difference spectroscopy, and the Na+-dependent build-up of a transmembrane voltage was demonstrated. NQR is a potential drug target for BV treatment since it is neither found in humans nor in Lactobacillus. In P. bivia, the highly active enzymes L-asparaginase and aspartate ammonia lyase catalyze the conversion of asparagine to the electron acceptor fumarate. However, the by-product ammonium is highly toxic. It has been proposed that P. bivia depends on ammonium-utilizing Gardnerella vaginalis, another typical pathogen associated with BV, and provides key nutrients to it. The product pattern of P. bivia growing on glucose in the presence of mixed amino acids substantiates this notion.
  • Publication
    Application of fluorescent proteins for functional dissection of the drosophila visual system
    (2021) Smylla, Thomas; Wagner, Krystina; Huber, Armin
    The Drosophila eye has been used extensively to study numerous aspects of biological systems, for example, spatio-temporal regulation of differentiation, visual signal transduction, protein trafficking and neurodegeneration. Right from the advent of fluorescent proteins (FPs) near the end of the millennium, heterologously expressed fusion proteins comprising FPs have been applied in Drosophila vision research not only for subcellular localization of proteins but also for genetic screens and analysis of photoreceptor function. Here, we summarize applications for FPs used in the Drosophila eye as part of genetic screens, to study rhodopsin expression patterns, subcellular protein localization, membrane protein transport or as genetically encoded biosensors for Ca2+ and phospholipids in vivo. We also discuss recently developed FPs that are suitable for super-resolution or correlative light and electron microscopy (CLEM) approaches. Illustrating the possibilities provided by using FPs in Drosophila photoreceptors may aid research in other sensory or neuronal systems that have not yet been studied as well as the Drosophila eye.
  • Publication
    Rickettsia spp. in ticks of South Luangwa valley, Eastern Province, Zambia
    (2023) Phiri, Bruno S. J.; Kattner, Simone; Chitimia-Dobler, Lidia; Woelfel, Silke; Albanus, Celina; Dobler, Gerhard; Küpper, Thomas
    Ticks are important vectors for Rickettsia spp. belonging to the Spotted Fever Group responsible for causing Rickettsiosis worldwide. Rickettsioses pose an underestimated health risk to tourists and local inhabitants. There is evidence of the presence of Rickettsia spp. in Zambia, however there is limited data. A total of 1465 ticks were collected in 20 different locations from dogs and cattle including one cat. Ticks were identified by morphological features or by sequencing of the 16S mitochondrial rRNA gene. Individual ticks were further tested for rickettsiae using a pan-Rickettsia real-time-PCR. Rickettsia species in PCR-positive ticks were identified by sequencing the 23S-5S intergenic spacer region or partial ompA gene, respectively. Seven tick species belonging to three different tick genera were found, namely: Amblyomma variegatum, Rhipicephalus appendiculatus, Rhipicephalus (Boophilus) microplus, Rhipicephalus simus, Rhipicephalus sanguineus, Rhipicephalus zambesiensis and Haemaphysalis elliptica. Out of the 1465 ticks collected, 67 (4.6%) tested positive in the pan-Rickettsia PCR. This study provides detailed data about the presence of Rickettsia species in South Luangwa Valley, Eastern Province, Zambia for the first time. High prevalence of Rickettsia africae in Amblyomma variegatum was found, which indicates the potential risk of infection in the investigated area. Furthermore, to our best knowledge, this is the first time Rickettsia massiliae, a human pathogen causing spotted fever, has been detected in Zambia.
  • Publication
    Serological protection rates against TBEV infection in blood donors from a highly endemic region in Southern Germany
    (2023) Dobler, Gerhard; Euringer, Kathrin; Kaier, Klaus; Borde, Johannes P.
    Background: Tick-borne encephalitis (TBE) is the most significant tick-borne disease in Europe and Asia, with more than 10,000 cases per year worldwide. A surge of reported TBE cases can be observed despite the availability of highly efficient vaccines. There is little known about the serological immune protection rate of the population in Germany. The seroprotection rate is defined as the presence of neutralizing antibodies. In contrast, the vaccination rate, as defined by public health agencies, may differ from the true protection rate in a population. Materials and Methods: 2220 blood samples from inhabitants of the county Ortenaukreis in the Federal State of Baden-Württemberg in Germany were included in the study. These were tested for anti-TBEV IgG antibodies by an anti-TBEV-IgG-ELISA. Subsequently, all TBEV-IgG positive samples were confirmed for neutralizing antibodies in the micro serum neutralization assay. Results: From the overall 2220 samples, 2104 were included in the comparison because of the selection of specific age groups (ages 20–69). In our sample size, we found an average serological protection rate (presence of neutralizing antibodies) of 57% (518/908) for the female blood donors and of 52% (632/1196) for the male blood donors. Discussion: In this study, we present new findings on a highly endemic region in southern Germany. Additionally, we present current data regarding the serological TBEV protection rates in the Ortenaukreis in southern Germany and compare these with a dataset published by the RKI, which is based on vaccination reports of the primary care providers and health care insurers, and with a self-reporting study conducted by a vaccine manufacturer. Our results significantly exceed the official numbers of average active vaccination status by 23.2% for females and by 21% for males. This might indicate an even longer persistence of TBE-vaccination-induced antibody titers than previously assumed.
  • Publication
    A high‐confidence Physcomitrium patens plasmodesmata proteome by iterative scoring and validation reveals diversification of cell wall proteins during evolution
    (2023) Gombos, Sven; Miras, Manuel; Howe, Vicky; Xi, Lin; Pottier, Mathieu; Kazemein Jasemi, Neda S.; Schladt, Moritz; Ejike, J. Obinna; Neumann, Ulla; Hänsch, Sebastian; Kuttig, Franziska; Zhang, Zhaoxia; Dickmanns, Marcel; Xu, Peng; Stefan, Thorsten; Baumeister, Wolfgang; Frommer, Wolf B.; Simon, Rüdiger; Schulze, Waltraud X.
    Plasmodesmata (PD) facilitate movement of molecules between plant cells. Regulation of this movement is still not understood. Plasmodesmata are hard to study, being deeply embedded within cell walls and incorporating several membrane types. Thus, structure and protein composition of PD remain enigmatic. Previous studies of PD protein composition identified protein lists with few validations, making functional conclusions difficult. We developed a PD scoring approach in iteration with large‐scale systematic localization, defining a high‐confidence PD proteome of Physcomitrium patens (HC300). HC300, together with bona fide PD proteins from literature, were placed in Pddb. About 65% of proteins in HC300 were not previously PD‐localized. Callose‐degrading glycolyl hydrolase family 17 (GHL17) is an abundant protein family with representatives across evolutionary scale. Among GHL17s, we exclusively found members of one phylogenetic clade with PD localization and orthologs occur only in species with developed PD. Phylogenetic comparison was expanded to xyloglucan endotransglucosylases/hydrolases and Exordium‐like proteins, which also diversified into PD‐localized and non‐PD‐localized members on distinct phylogenetic clades. Our high‐confidence PD proteome HC300 provides insights into diversification of large protein families. Iterative and systematic large‐scale localization across plant species strengthens the reliability of HC300 as basis for exploring structure, function, and evolution of this important organelle.
  • Publication
    Lipid remodeling of contrasting maize (Zea mays L.) hybrids under repeated drought
    (2023) Kränzlein, Markus; Schmöckel, Sandra M.; Geilfus, Christoph-Martin; Schulze, Waltraud X.; Altenbuchinger, Michael; Hrenn, Holger; Roessner, Ute; Zörb, Christian
    The role of recovery after drought has been proposed to play a more prominent role during the whole drought-adaption process than previously thought. Two maize hybrids with comparable growth but contrasting physiological responses were investigated using physiological, metabolic, and lipidomic tools to understand the plants’ strategies of lipid remodeling in response to repeated drought stimuli. Profound differences in adaptation between hybrids were discovered during the recovery phase, which likely gave rise to different degrees of lipid adaptability to the subsequent drought event. These differences in adaptability are visible in galactolipid metabolism and fatty acid saturation patterns during recovery and may lead to a membrane dysregulation in the sensitive maize hybrid. Moreover, the more drought-tolerant hybrid displays more changes of metabolite and lipid abundance with a higher number of differences within individual lipids, despite a lower physiological response, while the responses in the sensitive hybrid are higher in magnitude but lower in significance on the level of individual lipids and metabolites. This study suggests that lipid remodeling during recovery plays a key role in the drought response of plants.
  • Publication
    Increased vaccination diversity leads to higher and less-variable neutralization of TBE viruses of the European subtype
    (2023) Bestehorn-Willmann, Malena; Girl, Philipp; Greiner, Franziska; Mackenstedt, Ute; Dobler, Gerhard; Lang, Daniel
    Tick-borne encephalitis (TBE) is an infectious disease of the central nervous system. The causative agent is the tick-borne encephalitis virus (TBEV), which is most commonly transmitted by tick bites, but which may also be transmitted through the consumption of raw dairy products or, in rare instances, via infected transfusions, transplants, or the slaughter of infected animals. The only effective preventive option is active immunization. Currently, two vaccines are available in Europe—Encepur® and FSME-IMMUN®. In Central, Eastern, and Northern Europe, isolated TBEV genotypes belong mainly to the European subtype (TBEV-EU). In this study, we investigated the ability of these two vaccines to induce neutralizing antibodies against a panel of diverse natural TBEV-EU isolates from TBE-endemic areas in southern Germany and in regions of neighboring countries. Sera of 33 donors vaccinated with either FSME-IMMUN®, Encepur®, or a mixture of both were tested against 16 TBEV-EU strains. Phylogenetic analysis of the TBEV-EU genomes revealed substantial genetic diversity and ancestry of the identified 13 genotypic clades. Although all sera were able to neutralize the TBEV-EU strains, there were significant differences among the various vaccination groups. The neutralization assays revealed that the vaccination using the two different vaccine brands significantly increased neutralization titers, decreased intra-serum variance, and reduced the inter-virus variation.
  • Publication
    Industrial chicory genome gives insights into the molecular timetable of anther development and male sterility
    (2023) Waegneer, Evelien; Rombauts, Stephane; Baert, Joost; Dauchot, Nicolas; De Keyser, Annick; Eeckhaut, Tom; Haegeman, Annelies; Liu, Chang; Maudoux, Olivier; Notté, Christine; Staelens, Ariane; van der Veken, Jeroen; van Laere, Katrijn; Ruttink, Tom
    Industrial chicory (Cichorium intybus var. sativum) is a biannual crop mostly cultivated for extraction of inulin, a fructose polymer used as a dietary fiber. F1 hybrid breeding is a promising breeding strategy in chicory but relies on stable male sterile lines to prevent self-pollination. Here, we report the assembly and annotation of a new industrial chicory reference genome. Additionally, we performed RNA-Seq on subsequent stages of flower bud development of a fertile line and two cytoplasmic male sterile (CMS) clones. Comparison of fertile and CMS flower bud transcriptomes combined with morphological microscopic analysis of anthers, provided a molecular understanding of anther development and identified key genes in a range of underlying processes, including tapetum development, sink establishment, pollen wall development and anther dehiscence. We also described the role of phytohormones in the regulation of these processes under normal fertile flower bud development. In parallel, we evaluated which processes are disturbed in CMS clones and could contribute to the male sterile phenotype. Taken together, this study provides a state-of-the-art industrial chicory reference genome, an annotated and curated candidate gene set related to anther development and male sterility as well as a detailed molecular timetable of flower bud development in fertile and CMS lines.
  • Publication
    Detection and molecular characterization of canine babesiosis causative agent Babesia canis in naturally infected dogs in the Dobrogea area (Southeastern Romania)
    (2023) Ionita, Mariana; Leica, Laurentiu; Wassermann, Marion; Mitrea, Emanuel; Nicorescu, Isabela Madalina; Mitrea, Ioan Liviu
    Canine babesiosis is an emerging tick-borne disease of major veterinary concern in Europe. Its prevalence has increased in the last two decades and is spreading rapidly toward the north. The aim of this study was to investigate the genetic diversity of Babesia spp. strains isolated from naturally infected dogs in a tick-endemic area (Dobrogea) in southeastern Romania. For this purpose, a total of twenty-three samples from dogs diagnosed with various clinical forms of babesiosis, evaluated by means of clinical history, physical examination, and hematological tests, were subjected to a molecular investigation using PCR, sequencing analysis, and genetic characterization. A microscopic examination of thin Diff-quick-stained blood smears revealed large intra-erythrocytic Babesia piroplasms in all dogs. The PCR and sequencing analysis results indicated the presence of Babesia canis in 22 dogs (95.7%) and Babesia vogeli in 1 dog (4.3%). Among the B. canis isolates, two genotypes were distinguished based on two nucleotide substitutions (GA→AG) observed in the 18S rRNA gene sequences (at positions 609 and 610), with the AG genotype predominating (54.5% of samples), while the GA variant was identified in 9.1% of samples. In the remaining isolates (36.4%), both variants were identified. The B. vogeli-positive dog also tested positive for antibodies against Ehrlichia canis and displayed severe disease. This study reports, for the first time, the presence of genetically heterogenic B. canis strains in dogs with clinical babesiosis in Romania. These findings provide a basis for future studies on the relationship between the genetic structure of the causative agents of canine babesiosis in Romania and the course of the disease.
  • Publication
    Influence of ambient temperature on the phenology of the greater mouse‐eared bat (Myotis myotis)
    (2023) Matthäus, Laura; Kugelschafter, Karl; Fietz, Joanna
    In order to assess the consequences of climate change and evaluate its impacts on wildlife, it is essential to do so on a species-specific level. It is assumed that changes in the ambient temperature influence energy consumption as well as food availability and thus foraging behavior, reproduction, survival, and therefore population dynamics in bats. Based on this assumption, the present study aims to gain insights into the roosting and breeding behavior of the greater mouse-eared bat (Myotis myotis) in relation to changes of the ambient temperature. For this purpose, we investigated the effect of ambient temperature on the phenology of the greater mouse-eared bat by using activity data of the bats collected using light barriers at the maternity roosts. The light barrier used in this study is a system that detects the interruption of two light beams, for example, by a flying bat, and displays it as an electrical signal. The investigations have shown that 1. the higher the winter temperatures, the earlier the greater mouse-eared bats returned to the roosts to form the maternity colony; however, this was only true for ambient temperatures below 0.5°C, 2. birth season started earlier at higher spring temperatures, 3. the dissolution of maternity roosts occurred earlier with earlier birth season and at higher ambient temperatures during lactation. The results revealed that ambient temperature has an influence on the phenology of the greater mouse-eared bat. Depending on the respective life history stage, an increase in ambient temperature can have a positive or negative effect on the fitness of the animals. In recent years, mild winters have been recorded more frequently, which can have an influence on the behavior of bats. Warm winters within certain limits seem to lead to an earlier formation of the maternity colony, which can be positive or negative for the bats depending on persistent weather conditions and thus insect availability. In the course of climate change, we can also expect earlier spring events and an increase in spring temperature, as well as hot spells in summer. These warm springs and summers seem to lead to an earlier beginning of births, a faster development of the juveniles and an earlier dissolution of the maternity roost. An advance of reproductive activities can be assumed to increase the chance to survive the following winter in both mothers and their young, as they have more time to build up sufficient energy reserves for hibernation before winter starts. Due to the climatic changes, phenological changes of the bats be expected. This study highlights that in order to understand the impact of climate change on biodiversity, it is necessary to investigate in detail effects on a species-specific level and also to consider direct and indirect effects of ambient temperature on different life history stages.
  • Publication
    Helminths in invasive raccoons (Procyon lotor) from Southwest Germany
    (2023) Reinhardt, Nico P.; Wassermann, Marion; Härle, Jessica; Romig, Thomas; Kurzrock, Lina; Arnold, Janosch; Großmann, Ernst; Mackenstedt, Ute; Straubinger, Reinhard K.
    As hosts of numerous zoonotic pathogens, the role of raccoons needs to be considered in the One Health context. Raccoons progressively expand their range as invasive alien species in Europe. This study aimed to investigate the intestinal helminth fauna of raccoons in Baden-Wuerttemberg, Germany, as no such screening had ever been conducted there. In total, we obtained 102 animals from hunters in 2019 and 2020. Intestinal helminths were retrieved using the SSCT (segmented sedimentation and counting technique) and identified morphologically and by PCR-based Sanger sequencing. Fecal samples were assessed using the ELISA PetChekTM IP assay (IDEXX, Germany) and flotation technique. The artificial digestion method was employed for analyzing muscle tissue. We detected species of four nematode genera (Baylisascaris procyonis, Toxocara canis, Capillaria spp., and Trichuris spp.), three cestode genera (Atriotaenia cf. incisa/procyonis, Taenia martis, and Mesocestoides spp.), and three trematode genera (Isthmiophora hortensis/melis, Plagiorchis muris, and Brachylaima spp.). Echinococcus spp. and Trichinella spp. were not found. The invasive behavior and synanthropic habits of raccoons may increase the infection risk with these helminths in wildlife, domestic and zoo animals, and humans by serving as a connecting link. Therefore, it is crucial to initiate additional studies assessing these risks.
  • Publication
    The research process of PSK biosynthesis, signaling transduction, and potential applications in Brassica napus
    (2023) Shen, Xuwen; Stührwohldt, Nils; Lin, Chen
    Phytosulfokine (PSK) is a disulfated pentapeptide that acts as a growth regulator to control plant growth and development as well as adaptability to biotic and abiotic stress. In the last three decades, PSK has drawn increasing attention due to its various functions. Preproproteins that have been tyrosine sulfonylated and then cleaved by specific enzymes contribute to mature PSK. To transfer a signal from the apoplast to the inner cells, the PSK peptide must bind to the PSK receptors (PSKR1 and PSKR2) at the cell surface. The precise mechanism of PSK signal transduction is still unknown, given that PSKR combines receptor and kinase activity with a capacity to bind calmodulin (CaM). The binding of PSK and PSKR stimulates an abundance of cGMP downstream from PSKR, further activating a cation-translocating unit composed of cyclic nucleotide-gated channel 17 (CNGC17), H+-ATPases AHA1 and AHA2, and BRI-associated receptor kinase 1 (BAK1). Recently, it has been revealed that posttranslational ubiquitination is closely related to the control of PSK and PSKR binding. To date, the majority of studies related to PSK have used Arabidopsis. Given that rapeseed and Arabidopsis share a close genetic relationship, the relevant knowledge obtained from Arabidopsis can be further applied to rapeseed.
  • Publication
    New species based on the biological species concept within the complex of Lariophagus distinguendus (Hymenoptera, Chalcidoidea, Pteromalidae), a parasitoid of household pests
    (2023) Pollmann, Marie; Kuhn, Denise; König, Christian; Homolka, Irmela; Paschke, Sina; Reinisch, Ronja; Schmidt, Anna; Schwabe, Noa; Weber, Justus; Gottlieb, Yuval; Steidle, Johannes Luitpold Maria
    The pteromalid parasitoid Lariophagus distinguendus (Foerster) belongs to the Hymenoptera, a megadiverse insect order with high cryptic diversity. It attacks stored product pest beetles in human storage facilities. Recently, it has been shown to consist of two separate species. To further study its cryptic diversity, strains were collected to compare their relatedness using barcoding and nuclear genes. Nuclear genes identified two clusters which agree with the known two species, whereas the barcode fragment determined an additional third Clade. Total reproductive isolation (RI) according to the biological species concept (BSC) was investigated in crossing experiments within and between clusters using representative strains. Sexual isolation exists between all studied pairs, increasing from slight to strong with genetic distance. Postzygotic barriers mostly affected hybrid males, pointing to Haldane's rule. Hybrid females were only affected by unidirectional Spiroplasma‐induced cytoplasmic incompatibility and behavioural sterility, each in one specific strain combination. RI was virtually absent between strains separated by up to 2.8% COI difference, but strong or complete in three pairs from one Clade each, separated by at least 7.2%. Apparently, each of these clusters represents one separate species according to the BSC, highlighting cryptic diversity in direct vicinity to humans. In addition, these results challenge the recent ‘turbo‐taxonomy’ practice of using 2% COI differences to delimitate species, especially within parasitic Hymenoptera. The gradual increase in number and strength of reproductive barriers between strains with increasing genetic distance also sheds light on the emergence of barriers during the speciation process in L. distinguendus.
  • Publication
    DNA barcoding resolves quantitative multi‐trophic interaction networks and reveals pest species in trap nests
    (2023) Fornoff, Felix; Halla, Wenzel; Geiger, Sarah; Klein, Alexandra‐Maria; Sann, Manuela
    Insects, as one of the most species‐rich taxa with enormous taxonomic, behavioural and functional diversity, are in decline. Bees and wasps are especially crucial for ecosystems as pollinators or to control populations of other insects. To understand population drivers, comprehensive knowledge about top‐down and bottom‐up interactions, including all interaction partners, is needed. Nests of trap‐nesting bees and wasps include multi‐trophic interactions between bees, wasps, their food resources and natural enemies, simultaneously, however, up to today, all trophic interactions are not yet included in trap nest research because of challenges to identify the food used by nesting bees and wasps. Here, we reconstructed quantitative three‐ and four‐trophic interaction networks of species in three apoid wasp families using DNA barcoding. The obtained tripartite and quadripartite networks encompassed natural enemy‐wasp‐spider and natural enemy‐wasp‐herbivore‐plant interactions. Moreover, we identified so far undescribed Hymenoptera‐prey interactions, including prey species known as agricultural and forest pests. More extensive research on bee and wasp multitrophic interaction networks will provide valuable insights to better understand responses to environmental and biodiversity change, to investigate ecological theory and to reveal so far unknown feeding links.
  • Publication
    The Bacillus phage SPβ : a model system to study the lysis-lysogeny regulatory network and antiphage defense systems
    (2024) Kohm, Katharina; Commichau, Fabian M.
    Although bacteriophages are considered the most abundant biological entities on our planet, they are less well-studied compared to their host. Being intracellular parasites, phages rely on the metabolic processes of their bacterial hosts for their replication. Phages that use the host exclusively to produce virions are called virulent phages and the reproduction cycle is called the lytic cycle. The lytic cycle is accompanied by lysis and, thus, the killing of the host cell. Temperate phages can choose between the virulent or lysogenic lifecycle. Lysogeny or the lysogenic cycle is a type of viral reproduction in which no virus particles are produced, instead, the genetic material of the phage is replicated and then passed on to the daughter cells. The viral genome can be present as part of the bacterial chromosome or as a circular or linear plasmid molecule and is referred to as a prophage. Since temperate phages can influence the mutual interactions with other bacteria, growth, metabolic pathways or pathogenicity of their host, it is important to understand how temperate phages control their lysogenic life cycle and which genes are involved. Repression usually occurs through the interaction between a repressor and specific binding sites, which are mostly located in the promoter regions of the lytic genes. SPβ is a temperate phage of the model bacterium Bacillus subtilis. In contrast to its host, many aspects of the life cycle of SPβ have been little studied and many genes have not been assigned a function. Not only are SPβ-like phages widespread within the genus Bacillus and of greater importance to their hosts than previously thought, but they also exhibit a novel lysogeny management system. With regard to the control and regulation of the lysis-lysogeny network, it is already partially known which gene products are involved in the decision, establishment and resolvement of lysogeny. The maintenance and resolvement of lysogeny of SPβ was investigated in more detail in this thesis. To gain more insight into the regulation and control of lysogeny, the SPβ c2 mutant was characterized in this work. This mutant is unable to maintain its lysogenic state when exposed to heat, suggesting the alteration of a key regulatory element. This work demonstrated that the SPβ c2 phenotype is due to a single nucleotide exchange in the mrpR (yopR) gene that renders the encoded MrpRG136E protein temperature-sensitive. Furthermore, it was shown that this protein acts as a repressor of lytic gene expression. This occurs through the binding of the repressor to several conserved elements in the genome of the SPβ prophage. Further biochemical analysis revealed that the G136E exchange makes MrpR less stable and reduces its affinity for DNA binding. Structural characterization of MrpR revealed that the protein is a DNA-binding protein with a similar protein fold to tyrosine recombinases. However, the repressor function is independent of functional recombinase activity. In addition, a mutagenesis approach was used to identify the region within the protein that is essential for the function of the repressor. This work also identified further players in the lysogeny management system, with the YosL protein being crucial for the induction of the lytic cycle. However, YosL cannot activate the lytic cycle of SPβ alone. In addition, the core genome of SPβ-like phages was defined and new integration loci were identified in this work. Apart from a better understanding of lysis-lysogeny regulation and phagehost relationships, the characterization of the SPβ c2 mutant also led to the identification of a previously unknown phage defense system. The defense system is encoded on a plasmid and leads to a decrease in phage titer and a change in plaque morphology. It could be shown that the spbB locus, which ensures the segregation stability of the plasmid and codes for two open reading frames, is also responsible for the resistance to SPβ c2 and related phages. Further studies have shown that the spbB gene and the downstream region, which presumably encodes an RNA element and a terminator, play a crucial role in mediating resistance. The second open reading frame of the spbB locus is irrelevant for the mediation of phage resistance. Overall, this work contributes to a better understanding of the phage-host relationship.