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    Response of Collembola and Acari communities to summer flooding in a grassland plant diversity experiment 

    González-Macé, Odette; Scheu, Stefan
    PLOS ONE 2018; 13(8): Art. e0202862
    Flooding frequency is predicted to increase during the next decades in Europe. Therefore, it is important to understand how short-term disturbance events affect soil biota providing essential ecosystem functions and uncover factors modulating their response such as plant community composition. Here we report on the response of soil microarthropod communities (Collembola and Acari) to a severe summer flood in 2013, which affected major parts of central Europe. Collembola and Acari density and Collembola and Oribatida richness were strongly affected by the flood, but they recovered within three months. Effects of plant community composition on soil microarthropods disappeared after the flood, presumably due to homogenization of the field, but the effects of plant community were in a stage of being reasserted three months after the flood. Widespread, surface living and generalistic microarthropod species recolonized the field quickly. Prostigmata and Oribatida were more resilient or recovered to flooding than Astigmata and Gamasida. Long-term impacts, however, remain unknown and deserve further investigation.
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    Using imaging photoplethysmography for heart rate estimation in non-human primates 

    Unakafov, Anton M.; Möller, Sebastian; Kagan, Igor; Gail, Alexander; Treue, Stefan; Wolf, Fred
    PLOS ONE 2018; 13(8): Art. e0202581
    For humans and for non-human primates heart rate is a reliable indicator of an individual's current physiological state, with applications ranging from health checks to experimental studies of cognitive and emotional state. In humans, changes in the optical properties of the skin tissue correlated with cardiac cycles (imaging photoplethysmogram, iPPG) allow noncontact estimation of heart rate by its proxy, pulse rate. Yet, there is no established simple and non-invasive technique for pulse rate measurements in awake and behaving animals. Using iPPG, we here demonstrate that pulse rate in rhesus monkeys can be accurately estimated from facial videos. We computed iPPGs from eight color facial videos of four awake head-stabilized rhesus monkeys. Pulse rate estimated from iPPGs was in good agreement with reference data from a contact pulse-oximeter: the error of pulse rate estimation was below 5% of the individual average pulse rate in 83% of the epochs; the error was below 10% for 98% of the epochs. We conclude that iPPG allows non-invasive and non-contact estimation of pulse rate in non-human primates, which is useful for physiological studies and can be used toward welfare-assessment of non-human primates in research.
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    Functional diversity changes over 100 yr of primary succession on a volcanic island: insights into assembly processes 

    Karadimou, E.; Kallimanis, A. S.; Tsiripidis, I.; Raus, T.; Bergmeier, E.; Dimopoulos, P.
    Ecosphere 2018; 9(9): Art. e02374
    Changes in species diversity following volcanic eruptions have been studied extensively, but our knowledge on functional diversity and community assembly under such conditions is very limited. Here, we study the processes following the destruction of vegetation after a volcanic eruption. Specifically, we investigate (1) the temporal patterns of taxonomic and functional diversity over time since a previous eruption (alpha diversity) and beta diversity, (2) the temporal patterns of 26 individual traits (vegetative characteristics, plant taxa ecological preferences, and regenerative characteristics) providing more detailed information on species strategies at the initial and later stages of succession, and (3) the processes driving species assembly and whether they changed over time since the eruption an eruption. We analyzed data recorded during five floristic censuses that took place between 1911 and 2011, calculated alpha and beta facets of taxonomic and functional diversity and examined how community structure changed over time, using 26 functional characteristics, based on their ability to discern primary from later colonists, including longevity, growth form, Ellenberg’s indicator values, seed production and weight, flower size and sex, pollination type, and dispersal mode. Null model analysis was used to test whether the observed functional diversity deviates from random expectations. Alpha diversity, both taxonomic and functional, increased over time after an eruption, while beta diversity did not display a clear trend. This finding indicates that mainly abiotic processes determine species assembly over time after an eruption (at least for the time span studied here), contrary to theoretical expectations. It is most interesting that, simultaneously, some aspects of diversity indicated the effect of biotic interactions (facilitation and competition) on the assembly of species a few years after an eruption. This finding implies a legacy effect, since a high percentage of perennial species was noticed in the assemblage right after the eruption, as well as the effect of the harsh environmental conditions on the assembly of the plant communities. In conclusion, our results indicate the role of legacy effects in succession (most probably through the survival of underground plant parts) and underline the importance of disturbance history in providing the context needed for understanding effects of past events on succession.
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    How Rainforest Conversion to Agricultural Systems in Sumatra (Indonesia) Affects Active Soil Bacterial Communities 

    Berkelmann, Dirk; Schneider, Dominik; Engelhaupt, Martin; Heinemann, Melanie; Christel, Stephan; Wijayanti, Marini; Meryandini, Anja; Daniel, Rolf
    Frontiers in Microbiology 2018; 9: Art. 2381
    Palm oil production in Indonesia increased constantly over the last decades, which led to massive deforestation, especially on Sumatra island. The ongoing conversion of rainforest to agricultural systems results in high biodiversity loss. Here, we present the first RNA-based study on the effects of rainforest transformation to rubber and oil palm plantations in Indonesia for the active soil bacterial communities. For this purpose, bacterial communities of three different converted systems (jungle rubber, rubber plantation, and oil palm plantation) were studied in two landscapes with rainforest as reference by RT-PCR amplicon-based analysis of 16S rRNA gene transcripts. Active soil bacterial communities were dominated by Frankiales (Actinobacteria), subgroup 2 of the Acidobacteria and Alphaproteobacteria (mainly Rhizobiales and Rhodospirillales). Community composition differed significantly between the converted land use systems and rainforest reference sites. Alphaproteobacteria decreased significantly in oil palm samples compared to rainforest samples. In contrast, relative abundances of taxa within the Acidobacteria increased. Most important abiotic drivers for shaping soil bacterial communities were pH, calcium concentration, base saturation and C:N ratio. Indicator species analysis showed distinct association patterns for the analyzed land use systems. Nitrogen-fixing taxa including members of Rhizobiales and Rhodospirillales were associated with rainforest soils while nitrifiers and heat-resistant taxa including members of Actinobacteria were associated with oil palm soils. Predicted metabolic profiles revealed that the relative abundances of genes associated with fixation of nitrogen significantly decreased in plantation soils. Furthermore, predicted gene abundances regarding motility, competition or gene transfer ability indicated rainforest conversion-induced changes as well.
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    A perforated anodised aluminium slide for improved specimen clearing and imaging for confocal laser scanning microscopy 

    Quade, Felix S. C.; Preitz, Beate; Prpic, Nikola-Michael
    2018; 11(1): Art. 716
    Objective The bleaching, clearing and handling of tiny specimens with soft tissue and cuticular components for confocal laser scanning microscopy is difficult, because after cuticle bleaching and tissue clearing the specimens are virtually invisible. We have adjusted the design of the specimen container described by Smolla et al. (Arthropod Struct Dev 43:175–81, 2014) to handle tiny specimens. Results We describe a perforated and anodised aluminium slide that was designed to hold the distal tips of the pedipalp appendages of the spider Parasteatoda tepidariorum during clearing, and that can then be used directly for confocal laser scanning microscopy. We believe that this slide design will be helpful for others who want to visualise specimens between 500 and 800 µm with confocal laser scanning microscopy.
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    RAD sequencing resolved phylogenetic relationships in European shrub willows (Salix L. subg. Chamaetia and subg. Vetrix) and revealed multiple evolution of dwarf shrubs 

    Wagner, Natascha Dorothea; Gramlich, Susanne; Hörandl, Elvira
    Ecology and Evolution 2018; 8(16) p.8243-8255
    The large and diverse genus Salix L. is of particular interest for decades of biological research. However, despite the morphological plasticity, the reconstruction of phylogenetic relationships was so far hampered by the lack of informative molecular markers. Infrageneric classification based on morphology separates dwarf shrubs (subg. Chamaetia) and taller shrubs (subg. Vetrix), while previous phylogenetic studies placed species of these two subgenera just in one largely unresolved clade. Here we want to test the utility of genomic RAD sequencing markers for resolving relationships at different levels of divergence in Salix. Based on a sampling of 15 European species representing 13 sections of the two subgenera, we used five different RAD sequencing datasets generated by ipyrad to conduct phylogenetic analyses. Additionally we reconstructed the evolution of growth form and analyzed the genetic composition of the whole clade. The results showed fully resolved trees in both ML and BI analysis with high statistical support. The two subgenera Chamaetia and Vetrix were recognized as nonmonophyletic, which suggests that they should be merged. Within the Vetrix/Chamaetia clade, a division into three major subclades could be observed. All species were confirmed to be monophyletic. Based on our data, arctic-alpine dwarf shrubs evolved four times independently. The structure analysis showed five mainly uniform genetic clusters which are congruent in sister relationships observed in the phylogenies. Our study confirmed RAD sequencing as a useful genomic tool for the reconstruction of relationships on different taxonomic levels in the genus Salix.
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    Cattle selectivity by leopards suggests ways to mitigate human-leopard conflict 

    Khorozyan, Igor; Ghoddousi, Siavash; Soufi, Mobin; Soofi, Mahmood; Waltert, Matthias
    Ecology and Evolution 2018; 8(16) p.8011-8018
    Addressing widespread livestock losses to carnivores requires information on which livestock categories are preferentially selected. We analyzed an individual-based database of cattle grazing in forest (n = 932) and having been killed (n = 70) by leopards (Panthera pardus) in the Hyrcanian forest, Iran. We calculated Jacobs' selectivity index for cattle age, sex, and coloration across four scales: the study area as a whole, three sites, nine villages, and 60 cattle owners. Naturally colored cattle were significantly preferred by leopards at all scales in comparison with black and black-and-white cattle, and there was also a preference for males and juveniles at the study area level. More research is needed to see whether cattle losses would decrease if the share of naturally colored individuals in local holdings was reduced and males and juveniles had limited access to forest. We conclude that phenotypic and biologic characteristics of livestock can affect depredation and appeal for more research in this direction, particularly within the predator-prey framework.
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    Testing the plant pneumatic method to estimate xylem embolism resistance in stems of temperate trees 

    Zhang, Ya; Lamarque, Laurent J.; Torres-Ruiz, José M.; Schuldt, Bernhard; Karimi, Zohreh; Li, Shan; Qin, De-Wen; Bittencourt, Paulo; Burlett, Régis; Cao, Kun-Fang; et al.
    Delzon, SylvainOliveira, RafaelPereira, LucianoJansen, Steven
    Tree Physiology 2018; 38(7) p.1016-1025
    Methods to estimate xylem embolism resistance generally rely on hydraulic measurements, which can be far from straightforward. Recently, a pneumatic method based on air flow measurements of terminal branch ends was proposed to construct vulnerability curves by linking the amount of air extracted from a branch with the degree of embolism. We applied this novel technique for 10 temperate tree species, including six diffuse, two ring-porous and two gymnosperm species, and compared the pneumatic curves with hydraulic ones obtained from either the flow-centrifuge or the hydraulic-bench dehydration method. We found that the pneumatic method provides a good estimate of the degree of xylem embolism for all angiosperm species. The xylem pressure at 50% and 88% loss of hydraulic conductivity (i.e., Ψ50 and Ψ88) based on the methods applied showed a strongly significant correlation for all eight angiosperms. However, the pneumatic method showed significantly reduced Ψ50 values for the two conifers. Our findings suggest that the pneumatic method could provide a fast and accurate approach for angiosperms due to its convenience and feasibility, at least within the range of embolism resistances covered by our samples.
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    Velvet domain protein VosA represses the zinc cluster transcription factor SclB regulatory network for Aspergillus nidulans asexual development, oxidative stress response and secondary metabolism. 

    Thieme, Karl G.; Gerke, Jennifer; Sasse, Christoph; Valerius, Oliver; Thieme, Sabine; Karimi, Razieh; Heinrich, Antje K.; Finkernagel, Florian; Smith, Kristina; Bode, Helge B.; et al.
    Freitag, MichaelRam, Arthur F. J.Braus, Gerhard H.
    PLOS Genetics 2018; 14(7): Art. e1007511
    The NF-κB-like velvet domain protein VosA (viability of spores) binds to more than 1,500 promoter sequences in the filamentous fungus Aspergillus nidulans. VosA inhibits premature induction of the developmental activator gene brlA, which promotes asexual spore formation in response to environmental cues as light. VosA represses a novel genetic network controlled by the sclB gene. SclB function is antagonistic to VosA, because it induces the expression of early activator genes of asexual differentiation as flbC and flbD as well as brlA. The SclB controlled network promotes asexual development and spore viability, but is independent of the fungal light control. SclB interactions with the RcoA transcriptional repressor subunit suggest additional inhibitory functions on transcription. SclB links asexual spore formation to the synthesis of secondary metabolites including emericellamides, austinol as well as dehydroaustinol and activates the oxidative stress response of the fungus. The fungal VosA-SclB regulatory system of transcription includes a VosA control of the sclB promoter, common and opposite VosA and SclB control functions of fungal development and several additional regulatory genes. The relationship between VosA and SclB illustrates the presence of a convoluted surveillance apparatus of transcriptional control, which is required for accurate fungal development and the linkage to the appropriate secondary metabolism.
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    Crystal Structure of the Human tRNA Guanine Transglycosylase Catalytic Subunit QTRT1 

    Johannsson, Sven; Neumann, Piotr; Ficner, Ralf
    Biomolecules 2018; 8(3): Art. 81
    RNA modifications have been implicated in diverse and important roles in all kingdoms of life with over 100 of them present on tRNAs. A prominent modification at the wobble base of four tRNAs is the 7-deaza-guanine derivative queuine which substitutes the guanine at position 34. This exchange is catalyzed by members of the enzyme class of tRNA guanine transglycosylases (TGTs). These enzymes incorporate guanine substituents into tRNAAsp, tRNAAsn tRNAHis, and tRNATyr in all kingdoms of life. In contrast to the homodimeric bacterial TGT, the active eukaryotic TGT is a heterodimer in solution, comprised of a catalytic QTRT1 subunit and a noncatalytic QTRT2 subunit. Bacterial TGT enzymes, that incorporate a queuine precursor, have been identified or proposed as virulence factors for infections by pathogens in humans and therefore are valuable targets for drug design. To date no structure of a eukaryotic catalytic subunit is reported, and differences to its bacterial counterpart have to be deducted from sequence analysis and models. Here we report the first crystal structure of a eukaryotic QTRT1 subunit and compare it to known structures of the bacterial TGT and murine QTRT2. Furthermore, we were able to determine the crystal structure of QTRT1 in complex with the queuine substrate.
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    Comparative Genomics and Description of Putative Virulence Factors of Melissococcus plutonius, the Causative Agent of European Foulbrood Disease in Honey Bees 

    Djukic, Marvin; Erler, Silvio; Leimbach, Andreas; Grossar, Daniela; Charrière, Jean-Daniel; Gauthier, Laurent; Hartken, Denise; Dietrich, Sascha; Nacke, Heiko; Daniel, Rolf; et al.
    Poehlein, Anja
    Genes 2018; 9(8): Art. 419
    In Europe, approximately 84% of cultivated crop species depend on insect pollinators, mainly bees. Apis mellifera (the Western honey bee) is the most important commercial pollinator worldwide. The Gram-positive bacterium Melissococcus plutonius is the causative agent of European foulbrood (EFB), a global honey bee brood disease. In order to detect putative virulence factors, we sequenced and analyzed the genomes of 14 M. plutonius strains, including two reference isolates. The isolates do not show a high diversity in genome size or number of predicted protein-encoding genes, ranging from 2.021 to 2.101 Mbp and 1589 to 1686, respectively. Comparative genomics detected genes that might play a role in EFB pathogenesis and ultimately in the death of the honey bee larvae. These include bacteriocins, bacteria cell surface- and host cell adhesion-associated proteins, an enterococcal polysaccharide antigen, an epsilon toxin, proteolytic enzymes, and capsule-associated proteins. In vivo expression of three putative virulence factors (endo-alpha-N-acetylgalactosaminidase, enhancin and epsilon toxin) was verified using naturally infected larvae. With our strain collection, we show for the first time that genomic differences exist between non-virulent and virulent typical strains, as well as a highly virulent atypical strain, that may contribute to the virulence of M. plutonius. Finally, we also detected a high number of conserved pseudogenes (75 to 156) per genome, which indicates genomic reduction during evolutionary host adaptation.
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    Establishment of Apomixis in Diploid F2 Hybrids and Inheritance of Apospory From F1 to F2 Hybrids of the Ranunculus auricomus Complex 

    Barke, Birthe H.; Daubert, Mareike; Hörandl, Elvira
    Frontiers in Plant Science 2018; 9: Art. 1111
    Hybridization and polyploidization play important roles in plant evolution but it is still not fully clarified how these evolutionary forces contribute to the establishment of apomicts. Apomixis, the asexual reproduction via seed formation, comprises several essential alterations in development compared to the sexual pathway. Furthermore, most natural apomicts were found to be polyploids and/or hybrids. The Ranunculus auricomus complex comprises diploid sexual and polyploid apomictic species and represents an excellent model system to gain knowledge on origin and evolution of apomixis in natural plant populations. In this study, the second generation of synthetically produced homoploid (2x) and heteroploid (3x) hybrids derived from sexual R. auricomus species was analyzed for aposporous initial cell formation by DIC microscopy. Complete manifestation of apomixis was determined by measuring single mature seeds by flow cytometric seed screen. Microscopic analysis of the female gametophyte formation indicated spontaneous occurrence of aposporous initial cells and several developmental irregularities. The frequency of apospory was found to depend on dosage effects since a significant increase in apospory was observed, when both F1 parents, rather than just one, were aposporous. Other than in the F1 generation, diploid Ranunculus F2 hybrids formed BIII seeds and fully apomictic seeds. The results indicate that hybridization rather than polyploidization seems to be the functional activator of apomictic reproduction in the synthetic Ranunculus hybrids. In turn, at least two hybrid generations are required to establish apomictic seed formation.
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    Gender Differences in the Recognition of Vocal Emotions 

    Lausen, Adi; Schacht, Annekathrin
    Frontiers in Psychology 2018; 9: Art. 882
    The conflicting findings from the few studies conducted with regard to gender differences in the recognition of vocal expressions of emotion have left the exact nature of these differences unclear. Several investigators have argued that a comprehensive understanding of gender differences in vocal emotion recognition can only be achieved by replicating these studies while accounting for influential factors such as stimulus type, gender-balanced samples, number of encoders, decoders, and emotional categories. This study aimed to account for these factors by investigating whether emotion recognition from vocal expressions differs as a function of both listeners' and speakers' gender. A total of N = 290 participants were randomly and equally allocated to two groups. One group listened to words and pseudo-words, while the other group listened to sentences and affect bursts. Participants were asked to categorize the stimuli with respect to the expressed emotions in a fixed-choice response format. Overall, females were more accurate than males when decoding vocal emotions, however, when testing for specific emotions these differences were small in magnitude. Speakers' gender had a significant impact on how listeners' judged emotions from the voice. The group listening to words and pseudo-words had higher identification rates for emotions spoken by male than by female actors, whereas in the group listening to sentences and affect bursts the identification rates were higher when emotions were uttered by female than male actors. The mixed pattern for emotion-specific effects, however, indicates that, in the vocal channel, the reliability of emotion judgments is not systematically influenced by speakers' gender and the related stereotypes of emotional expressivity. Together, these results extend previous findings by showing effects of listeners' and speakers' gender on the recognition of vocal emotions. They stress the importance of distinguishing these factors to explain recognition ability in the processing of emotional prosody.
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    Heavy Metal-Induced Expression of PcaA Provides Cadmium Tolerance to Aspergillus fumigatus and Supports Its Virulence in the Galleria mellonella Model 

    Bakti, Fruzsina; Sasse, Christoph; Heinekamp, Thorsten; Pócsi, István; Braus, Gerhard H.
    Frontiers in Microbiology 2018; 9: Art. 744
    Most of the metal transporters in Aspergillus fumigatus are yet uncharacterized. Their role in fungal metabolism and virulence remains unclear. This paper describes the novel PIB-type cation ATPase PcaA, which links metal homeostasis and heavy metal tolerance in the opportunistic human pathogen A. fumigatus. The protein possesses conserved ATPase motif and shares 51% amino acid sequence identity with the Saccharomyces cerevisiae cadmium exporter Pca1p. A pcaA deletion, an overexpression and a gfp-pcaA complementation strain of A. fumigatus were constructed and their heavy metal susceptibilities were studied. The pcaA knock out strain showed drastically decreased cadmium tolerance, however, its growth was not affected by the exposure to high concentrations of copper, iron, zinc, or silver ions. Although the lack of PcaA had no effect on copper adaption, we demonstrated that not only cadmium but also copper ions are able to induce the transcription of pcaA in A. fumigatus wild type Af293. Similarly, cadmium and copper ions could induce the copper exporting ATPase crpA. These data imply a general response on the transcriptomic level to heavy metals in A. fumigatus through the induction of detoxification systems. Confocal microscopy of the gfp-pcaA complementation strain expressing functional GFP-PcaA supports the predicted membrane localization of PcaA. The GFP-PcaA fusion protein is located in the plasma membrane of A. fumigatus in the presence of cadmium ions. Virulence assays support a function of PcaA for virulence of A. fumigatus in the Galleria mellonella wax moth larvae model, which might be linked to the elimination of reactive oxygen species.
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    Changes in the microstructure of compact and trabecular bone tissues of mice subchronically exposed to alcohol 

    Martiniakova, Monika; Sarocka, Anna; Babosova, Ramona; Grosskopf, Birgit; Kapusta, Edyta; Goc, Zofia; Formicki, Grzegorz; Omelka, Radoslav
    2018; 25(1): Art. 8
    Background: Alcohol is one of the most commonly consumed neurotoxins by humans. Its negative effect on bone health is known for a long time. However, its impact on qualitative and quantitative 2D characteristics of the compact bone is still unclear. Therefore, the aim of this study was to investigate in detail the effects of subchronic alcohol exposure on compact and trabecular bone tissues microstructure of laboratory mice using 2D and 3D imaging methods. Ten clinically healthy 12 weeks-old mice (males) were randomly divided into two groups. Animals from experimental group (group E; n = 5) drank a solution composed of 15% ethanol and water (1.7 g 100% ethanol kg-1 b.w. per day) for 8 weeks, while those from control group (group C; n = 5) drank only water. Results: Subchronic exposure to alcohol leads to several changes in qualitative 2D characteristics of the compact bone such as the presence of primary vascular radial bone tissue in pars anterior of endosteal border and a higher number of resorption lacunae (five times more) in the middle part of substantia compacta. Morphometrical 2D evaluations of the compact bone showed significantly increased sizes of primary osteons' vascular canals (p < 0.05) in mice from the experimental group (E group). Sizes of Haversian canals and secondary osteons were not affected by alcohol consumption. In mice from the E group, significantly lower values for relative bone volume and bone mineral density of the compact bone were observed. In the trabecular bone, decreased values for bone volume, trabecular number, trabecular thickness and bone surface (p < 0.05) were documented. Conclusions: Alcohol decreased not only bone volume and density of the compact bone, but it also reduced trabecular bone volume and leads to trabecular thinning. It caused vasodilation of primary osteons' vascular canals and increased porosity in the compact bone.
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    A CRISPR-Cas9-Based Toolkit for Fast and Precise In Vivo Genetic Engineering of Bacillus subtilis Phages 

    Schilling, Tobias; Dietrich, Sascha; Hoppert, Michael; Hertel, Robert
    Viruses 2018; 10(5): Art. 241
    Phages are currently under discussion as a solution for the antibiotic crisis, as they may cure diseases caused by multi-drug-resistant pathogens. However, knowledge of phage biology and genetics is limited, which impedes risk assessment of therapeutic applications. In order to enable advances in phage genetic research, the aim of this work was to create a toolkit for simple and fast genetic engineering of phages recruiting Bacillus subtilis as host system. The model organism B. subtilis represents a non-pathogenic surrogate of its harmful relatives, such as Bacillus anthracis or Bacillus cereus. This toolkit comprises the application CutSPR, a bioinformatic tool for rapid primer design, and facilitates the cloning of specific CRISPR-Cas9-based mutagenesis plasmids. The employment of the prophage-free and super-competent B. subtilis TS01 strain enables an easy and fast introduction of specific constructs for in vivo phage mutagenesis. Clean gene deletions and a functional clean gene insertion into the genome of the model phage vB_BsuP-Goe1 served as proof of concept and demonstrate reliability and high efficiency. The here presented toolkit allows comprehensive investigation of the diverse phage genetic pool, a better understanding of phage biology, and safe phage applications.
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    Reconstructing geographical parthenogenesis: effects of niche differentiation and reproductive mode on Holocene range expansion of an alpine plant. 

    Kirchheimer, Bernhard; Wessely, Johannes; Gattringer, Andreas; Hülber, Karl; Moser, Dietmar; Schinkel, Christoph C. F.; Appelhans, Marc; Klatt, Simone; Caccianiga, Marco; Dellinger, Agnes; et al.
    Guisan, AntoineKuttner, MichaelLenoir, JonathanMaiorano, LuigiNieto-Lugilde, DiegoPlutzar, ChristophSvenning, Jens-ChristianWillner, WolfgangHörandl, ElviraDullinger, Stefan
    Ecology Letters 2018; 21(3) p.392-401
    Asexual taxa often have larger ranges than their sexual progenitors, particularly in areas affected by Pleistocene glaciations. The reasons given for this 'geographical parthenogenesis' are contentious, with expansion of the ecological niche or colonisation advantages of uniparental reproduction assumed most important in case of plants. Here, we parameterized a spread model for the alpine buttercup Ranunculus kuepferi and reconstructed the joint Holocene range expansion of its sexual and apomictic cytotype across the European Alps under different simulation settings. We found that, rather than niche broadening or a higher migration rate, a shift of the apomict's niche towards colder conditions per se was crucial as it facilitated overcoming of topographical barriers, a factor likely relevant for many alpine apomicts. More generally, our simulations suggest potentially strong interacting effects of niche differentiation and reproductive modes on range formation of related sexual and asexual taxa arising from their differential sensitivity to minority cytotype disadvantage.
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    A toolkit for Nannochloropsis oceanica CCMP1779 enables gene stacking and genetic engineering of the eicosapentaenoic acid pathway for enhanced long-chain polyunsaturated fatty acid production 

    Poliner, Eric; Pulman, Jane A.; Zienkiewicz, Krzysztof; Childs, Kevin; Benning, Christoph; Farré, Eva M.
    Plant Biotechnology Journal 2018; 16(1) p.298-309
    Nannochloropsis oceanica is an oleaginous microalga rich in ω3 long-chain polyunsaturated fatty acids (LC-PUFAs) content, in the form of eicosapentaenoic acid (EPA). We identified the enzymes involved in LC-PUFA biosynthesis in N. oceanica CCMP1779 and generated multigene expression vectors aiming at increasing LC-PUFA content in vivo. We isolated the cDNAs encoding four fatty acid desaturases (FAD) and determined their function by heterologous expression in S. cerevisiae. To increase the expression of multiple fatty acid desaturases in N. oceanica CCMP1779, we developed a genetic engineering toolkit that includes an endogenous bidirectional promoter and optimized peptide bond skipping 2A peptides. The toolkit also includes multiple epitopes for tagged fusion protein production and two antibiotic resistance genes. We applied this toolkit, towards building a gene stacking system for N. oceanica that consists of two vector series, pNOC-OX and pNOC-stacked. These tools for genetic engineering were employed to test the effects of the overproduction of one, two or three desaturase-encoding cDNAs in N. oceanica CCMP1779 and prove the feasibility of gene stacking in this genetically tractable oleaginous microalga. All FAD overexpressing lines had considerable increases in the proportion of LC-PUFAs, with the overexpression of Δ12 and Δ5 FAD encoding sequences leading to an increase in the final ω3 product, EPA.
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    Cyanophage-encoded lipid desaturases: oceanic distribution, diversity and function 

    Roitman, Sheila; Hornung, Ellen; Flores-Uribe, José; Sharon, Itai; Feussner, Ivo; Béjà, Oded
    The ISME Journal 2018; 12(2) p.343-355
    Cyanobacteria are among the most abundant photosynthetic organisms in the oceans; viruses infecting cyanobacteria (cyanophages) can alter cyanobacterial populations, and therefore affect the local food web and global biochemical cycles. These phages carry auxiliary metabolic genes (AMGs), which rewire various metabolic pathways in the infected host cell, resulting in increased phage fitness. Coping with stress resulting from photodamage appears to be a central necessity of cyanophages, yet the overall mechanism is poorly understood. Here we report a novel, widespread cyanophage AMG, encoding a fatty acid desaturase (FAD), found in two genotypes with distinct geographical distribution. FADs are capable of modulating the fluidity of the host's membrane, a fundamental stress response in living cells. We show that both viral FAD (vFAD) families are Δ9 lipid desaturases, catalyzing the desaturation at carbon 9 in C16 fatty acid chains. In addition, we present a comprehensive fatty acid profiling for marine cyanobacteria, which suggests a unique desaturation pathway of medium- to long-chain fatty acids no longer than C16, in accordance with the vFAD activity. Our findings suggest that cyanophages are capable of fiddling with the infected host's membranes, possibly leading to increased photoprotection and potentially enhancing viral-encoded photosynthetic proteins, resulting in a new viral metabolic network.
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    Dendrochronologically dated pine stumps document phase-wise bog expansion at a northwest German site between ca. 6700 and ca. 3400 BC 

    Achterberg, Inke Elisabeth Maike; Eckstein, Jan; Birkholz, Bernhard; Bauerochse, Andreas; Leuschner, Hanns Hubert
    Climate of the Past 2018; 14(1) p.85-100
    The investigated northwest German mire site at "Totes Moor" is densely covered with subfossil pine stumps (Pinus sylvestris L.) from the fen–bog transition. This facilitates the spatio-temporal reconstruction of mire development, which is based on 212 in situ tree stumps in the case study presented here. Six dendrochronologically dated site chronologies together cover 2345 years between 6703 and 3403 BC. The gaps in between are 6 to 550 years long. Additionally, a floating chronology of 309 years, containing 30 trees, was radiocarbon-dated to the beginning of the 7th millennium cal BC. Peat-stratigraphical survey was carried out additionally, and elevations a.s.l. were determined at several locations. Tree dying-off phases, which indicate water level rise at the site, mostly in context of the local fen–bog transition, are evident for ca. 6600–6450, ca. 6350–5750, ca. 5300–4900, ca. 4700–4550, ca. 3900–3850, ca. 3700–3600, ca. 3500–3450 and ca. 3400 BC. The spatial distribution of the dated in situ trees illustrates the phase-wise expansion of raised bog over fen peat at the site. The documented bog expansion pulses likely correspond to climatic wet sifts.
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