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Acrocomia aculeata fruits from three regions in Costa Rica: An assessment of biometric parameters, oil content and oil fatty acid composition to evaluate industrial potential
(2020) Alfaro-Solís, Jose David; Montoya-Arroyo, Alexander; Jiménez, Víctor M.; Arnáez-Serrano, Elizabeth; Pérez, Jason; Vetter, Walter; Frank, Jan; Lewandowski, Iris
Due to increased global demand for vegetable oils, diversification of the supply chain with sustainable sources is necessary. Acrocomia aculeata has recently gained attention as a multi-purpose, sustainable crop for oil production. However, the information necessary for effective selection of promising varieties for agricultural production is lacking. The aim of this study was to assess variability in fruit morphology and oil composition of individual Acrocomia aculeata plants growing wild in different climatic regions of Costa Rica. Fruits at the same ripening stage were collected at three locations, and biometric features, oil content, fatty acid composition of oils from kernels and pulp, as well as fiber composition of husks were determined. Biometric parameters showed high variability among the regions assessed. Moreover, oil content and relative proportions of unsaturated fatty acids were higher at the most tropical location, whereas lauric acid content was lowest under these conditions, indicating a potential environmental effect on oil composition. Pulp oil content correlated positively with annual precipitation and relative humidity, but no clear relation to temperature was observed. The oil chemical composition was similar to that reported for Elaeis guineensis, suggesting that Acrocomia aculeata from Costa Rica may be a suitable alternative for industrial applications currently based on African palm oil. Analysis of husks as a coproduct revealed the possibility of obtaining materials with high lignin and low water and ash contents that could be used as a solid bioenergy source. In conclusion, Acrocomia aculeata oil is a promising alternative for industrial applications currently based on African palm oil and byproducts of its oil production could find additional use as a renewable energy source.
The antioxidant potential of various wheat crusts correlates with AGE content independently of acrylamide
(2023) Wächter, Kristin; Longin, Carl Friedrich H.; Winterhalter, Patrick R.; Bertsche, Ute; Szabó, Gábor; Simm, Andreas
Epidemiological studies have indicated that the consumption of whole-grain products is associated with a reduced risk of cardiovascular diseases, type II diabetes, and cancer. In the case of bread, high amounts of antioxidants and advanced glycation end products (AGEs) are formed during baking by the Maillard reaction in the bread crust; however, the formation of potentially harmful compounds such as acrylamide also occurs. This study investigated the antioxidant responses of different soluble extracts from whole-grain wheat bread crust extracts (WBCEs) in the context of the asparagine, AGE, and acrylamide content. For that, we analyzed nine bread wheat cultivars grown at three different locations in Germany (Hohenheim, Eckartsweier, and Oberer Lindenhof). We determined the asparagine content in the flour of the 27 wheat cultivars and the acrylamide content in the crust, and measured the antioxidant potential using the induced expression of the antioxidant genes GCLM and HMOX1 in HeLa cells. Our study uncovered, for the first time, that the wheat crust’s antioxidant potential correlates with the AGE content, but not with the acrylamide content. Mass spectrometric analyses of WBCEs for identifying AGE-modified proteins relevant to the antioxidant potential were unsuccessful. However, we did identify the wheat cultivars with a high antioxidant potential while forming less acrylamide, such as Glaucus and Lear. Our findings indicate that the security of BCEs with antioxidative and cardioprotective potential can be improved by choosing the right wheat variety.
Appreciable genetic correlation between inbred lines and testcrosses facilitates breeding for resistance to Fusarium head blight in hybrid rye (Secale cereale)
(2021) Miedaner, Thomas; Rose, Marvin; Overbeck, Franziska; Koch, Silvia; Gruner, Paul; Eifler, Jakob
Hybrid breeding is based on selection of inbred lines in early generations. A sufficient genetic correlation between inbred lines and testcrosses is, however, the prerequisite for a higher selection gain in the hybrids. Therefore, we investigated this crucial parameter for Fusarium head blight (FHB) resistance on 60 inbred lines each of the heterotic groups Petkus and Carsten and their corresponding crosses with two testers each at four (inbred lines) or six (testcrosses) environments (location × year combinations). FHB severity measured as percentage of infected spikelets per plot was used as resistance trait, and a correction was made by using flowering time as covariate. Variances for genotype and genotype–environment interaction were high, and the heritability was estimated .84 and .74 for the line per se performance (Petkus and Carsten) and .71 and .78 for the general combining ability (GCA). For both inbred lines and testcrosses, negative correlations with plant height were found ranging from −.20 to −.51. FHB severity was considerably higher for the inbred lines than for the testcrosses. Genetic variance was lower for testcrosses, and dominance effects of the testers and heterosis may be key factors reducing the general infection level of three‐way hybrids. The effect of the tester (i.e., difference between testers) was very small and only significant for the Carsten group. The variance for specific combining ability (SCA) was very small, too. This, and high genetic correlations between line per se and GCA (r = .82 for Petkus and r = .72 for Carsten), does allow or even favour the selection based on line per se performance. Still, in the last generation before the official trials, testcrosses should also be tested for FHB resistance and mycotoxin contents to reliably improve food and feed quality in rye.
Bioprocess exploitation of microaerobic auto-induction using the example of rhamnolipid biosynthesis in Pseudomonas putida KT2440
(2025) Grether, Jakob; Dittmann, Holger; Willems, Leon; Schmiegelt, Tabea; Benatto Perino, Elvio Henrique; Hubel, Philipp; Lilge, Lars; Hausmann, Rudolf
Background: In biomanufacturing of surface-active agents, such as rhamnolipids, excessive foaming is a significant obstacle for the development of high-performing bioprocesses. The exploitation of the inherent tolerance of Pseudomonas putida KT2440, an obligate aerobic bacterium, to microaerobic conditions has received little attention so far. Here low-oxygen inducible promoters were characterized in biosensor strains and exploited for process control under reduction of foam formation by low aeration and stirring rates during biosynthesis of rhamnolipids. Results: In this study, homologous promoters of P. putida inducible under oxygen limitation were identified by non-targeted proteomic analyses and characterized by fluorometric methods. Proteomics indicated a remodeling of the respiratory chain and the regulation of stress-related proteins under oxygen limitation. Of the three promoters tested in fluorescent biosensor assays, the promoter of the oxygen-sensitive cbb3-type cytochrome c oxidase gene showed high oxygen-dependent controllability. It was used to control the gene expression of a heterologous di-rhamnolipid synthesis operon in an auto-inducing microaerobic two-phase bioprocess. By limiting the oxygen supply via low aeration and stirring rates, the bioprocess was clearly divided into a growth and a production phase, and sources of foam formation were reduced. Accordingly, rhamnolipid synthesis did not have to be controlled externally, as the oxygen-sensitive promoter was autonomously activated as soon as the oxygen level reached microaerobic conditions. A critical threshold of about 20% oxygen saturation was determined. Conclusions: Utilizing the inherent tolerance of P. putida to microaerobic conditions in combination with the application of homologous, low-oxygen inducible promoters is a novel and efficient strategy to control bioprocesses. Fermentation under microaerobic conditions enabled the induction of rhamnolipid production by low oxygen levels, while foam formation was limited by low aeration and stirring rates.
Breeding progress of disease resistance and impact of disease severity under natural infections in winter wheat variety trials
(2021) Laidig, F.; Feike, T.; Hadasch, S.; Rentel, D.; Klocke, B.; Miedaner, T.; Piepho, H. P.
Key message: Breeding progress of resistance to fungal wheat diseases and impact of disease severity on yield reduction in long-term variety trials under natural infection were estimated by mixed linear regression models. Abstract: This study aimed at quantifying breeding progress achieved in resistance breeding towards varieties with higher yield and lower susceptibility for 6 major diseases, as well as estimating decreasing yields and increasing disease susceptibility of varieties due to ageing effects during the period 1983–2019. A further aim was the prediction of disease-related yield reductions during 2005–2019 by mixed linear regression models using disease severity scores as covariates. For yield and all diseases, overall progress of the fully treated intensity (I2) was considerably higher than for the intensity without fungicides and growth regulators (I1). The disease severity level was considerably reduced during the study period for mildew (MLD), tan spot (DTR) and Septoria nodorum blotch (ear) (SNB) and to a lesser extent for brown (leaf) rust (BNR) and Septoria tritici blotch (STB), however, not for yellow/stripe rust (YLR). Ageing effects increased susceptibility of varieties strongly for BNR and MLD, but were comparatively weak for SNB and DTR. Considerable yield reductions under high disease severity were predicted for STB (−6.6%), BNR (−6.5%) and yellow rust (YLR, −5.8%), but lower reductions for the other diseases. The reduction for resistant vs. highly susceptible varieties under high severity conditions was about halved for BNR and YLR, providing evidence of resistance breeding progress. The empirical evidence on the functional relations between disease severity, variety susceptibility and yield reductions based on a large-scale multiple-disease field trial data set in German winter wheat is an important contribution to the ongoing discussion on fungicide use and its environmental impact.
Can we abandon phosphorus starter fertilizer in maize? Results from a diverse panel of elite and doubled haploid landrace lines of maize (Zea mays L.)
(2022) Roller, Sandra; Weiß, Thea M.; Li, Dongdong; Liu, Wenxin; Schipprack, Wolfgang; Melchinger, Albrecht E.; Hahn, Volker; Leiser, Willmar L.; Würschum, Tobias
The importance of phosphorus (P) in agriculture contrasts with the negative environmental impact and the limited resources worldwide. Reducing P fertilizer application by utilizing more efficient genotypes is a promising way to address these issues. To approach this, a large panel of maize (Zea mays L.) comprising each 100 Flint and Dent elite lines and 199 doubled haploid lines from six landraces was assessed in multi-environment field trials with and without the application of P starter fertilizer. The treatment comparison showed that omitting the starter fertilizer can significantly affect traits in early plant development but had no effect on grain yield. Young maize plants provided with additional P showed an increased biomass, faster growth and superior vigor, which, however, was only the case under environmental conditions considered stressful for maize cultivation. Importantly, though the genotype-by-treatment interaction variance was comparably small, there is genotypic variation for this response that can be utilized in breeding. The comparison of elite and doubled haploid landrace lines revealed a superior agronomic performance of elite material but also potentially valuable variation for early traits in the landrace doubled haploid lines. In conclusion, our results illustrate that breeding for P efficient maize cultivars is possible towards a reduction of P fertilizer in a more sustainable agriculture.
Changes of microorganism composition in fresh and stored bee pollen from Southern Germany
(2021) Friedle, Carolin; D’Alvise, Paul; Schweikert, Karsten; Wallner, Klaus; Hasselmann, Martin
Analysis of plant pollen can provide valuable insights into the existing spectrum of microorganisms in the environment. When harvesting bee-collected pollen as a dietary supplement for human consumption, timely preservation of the freshly collected pollen is fundamental for product quality. Environmental microorganisms contained in freshly collected pollen can lead to spoilage by degradation of pollen components. In this study, freshly collected bee pollen was sampled at different locations and stored under various storage conditions to examine the hypothesis that storage conditions may have an effect on the composition of microorganisms in pollen samples. The samples were analyzed using 16S and 18S amplicon sequencing and characterized by palynological analysis. Interestingly, the bacterial communities between pollen samples from different locations varied only slightly, whereas for fungal community compositions, this effect was substantially increased. Further, we noticed that fungal communities in pollen are particularly sensitive to storage conditions. The fungal genera proportion Cladosporium and Mycosphaerella decreased, while Zygosaccharomyces and Aspergillus increased during storage. Aspergillus and Zygosaccharomyces fractions increased during storage at 30 °C, which could negatively impact the pollen quality if it is used as a dietary supplement.
Characterization of Bacillus velezensis UTB96, demonstrating improved lipopeptide production compared to the strain B. velezensis FZB42
(2022) Vahidinasab, Maliheh; Adiek, Isabel; Hosseini, Behnoush; Akintayo, Stephen Olusanmi; Abrishamchi, Bahar; Pfannstiel, Jens; Henkel, Marius; Lilge, Lars; Vögele, Ralf ; Hausmann, Rudolf
Bacillus strains can produce various lipopeptides, known for their antifungal properties. This makes them attractive metabolites for applications in agriculture. Therefore, identification of productive wild-type strains is essential for the development of biopesticides. Bacillus velezensis FZB42 is a well-established strain for biocontrol of plant pathogens in agriculture. Here, we characterized an alternative strain, B. velezensis UTB96, that can produce higher amounts of all three major lipopeptide families, namely surfactin, fengycin, and iturin. UTB96 produces iturin A. Furthermore, UTB96 showed superior antifungal activity towards the soybean fungal pathogen Diaporthe longicolla compared to FZB42. Moreover, the additional provision of different amino acids for lipopeptide production in UTB96 was investigated. Lysine and alanine had stimulatory effects on the production of all three lipopeptide families, while supplementation of leucine, valine and isoleucine decreased the lipopeptide bioproduction. Using a 45-litre bioreactor system for upscaling in batch culture, lipopeptide titers of about 140 mg/L surfactin, 620 mg/L iturin A, and 45 mg/L fengycin were achieved. In conclusion, it becomes clear that B. velezensis UTB96 is a promising strain for further research application in the field of agricultural biological controls of fungal diseases.
Characterization ofantifungal properties of lipopeptide-producing Bacillus velezensis strains and their proteome-based response to the phytopathogens, Diaporthe spp
(2023) Akintayo, Stephen Olusanmi; Hosseini, Behnoush; Vahidinasab, Maliheh; Messmer, Marc; Pfannstiel, Jens; Bertsche, Ute; Hubel, Philipp; Henkel, Marius; Hausmann, Rudolf; Vögele, Ralf; Lilge, Lars
Introduction: B. velezensis strains are of interest in agricultural applications due to their beneficial interactions with plants, notable through their antimicrobial activity. The biocontrol ability of two new lipopeptides-producing B. velezensis strains ES1-02 and EFSO2-04, against fungal phytopathogens of Diaporthe spp., was evaluated and compared with reference strains QST713 and FZB42. All strains were found to be effective against the plant pathogens, with the new strains showing comparable antifungal activity to QST713 and slightly lower activity than FZB42. Methods: Lipopeptides and their isoforms were identified by high-performance thin-layer chromatography (HPTLC) and mass spectrometric measurements. The associated antifungal influences were determined in direct in vitro antagonistic dual culture assays, and the inhibitory growth effects on Diaporthe spp. as representatives of phytopathogenic fungi were determined. The effects on bacterial physiology of selected B. velezensis strains were analyzed by mass spectrometric proteomic analyses using nano-LC-MS/MS. Results and Discussion: Lipopeptide production analysis revealed that all strains produced surfactin, and one lipopeptide of the iturin family, including bacillomycin L by ES1-02 and EFSO2-04, while QST713 and FZB42 produced iturin A and bacillomycin D, respectively. Fengycin production was however only detected in the reference strains. As a result of co-incubation of strain ES1-02 with the antagonistic phytopathogen D. longicolla, an increase in surfactin production of up to 10-fold was observed, making stress induction due to competitors an attractive strategy for surfactin bioproduction. An associated global proteome analysis showed a more detailed overview about the adaptation and response mechanisms of B. velezensis, including an increased abundance of proteins associated with the biosynthesis of antimicrobial compounds. Furthermore, higher abundance was determined for proteins associated with oxidative, nitrosative, and general stress response. In contrast, proteins involved in phosphate uptake, amino acid transport, and translation were decreased in abundance. Altogether, this study provides new insights into the physiological adaptation of lipopeptide-producing B. velezensis strains, which show the potential for use as biocontrol agents with respect to phytopathogenic fungi.
Chemometric approach for profiling of metabolites of potential antioxidant activity in Apiaceae species based on LC-PDA-ESI-MS/MS and FT-NIR
(2023) Atta, Noha H.; Handoussa, Heba; Klaiber, Iris; Hitzmann, Bernd; Hanafi, Rasha S.
Chemometrics is a tool for data mining and unlocking the door for solving big data queries. Apiaceae is a family species which is commonly cultivated worldwide. Although members of this species are widely used as antioxidant, antibacterial, antifungal, and anti-inflammatory agents, their metabolites profiling remains ambiguous. Based on WHO support, chemometrics has been used in evaluating the quality and authenticity of the herbal products. The objective of this study is to profile and characterize phenolic metabolites in nine species from Egyptian cultivars and three different species of German cultivars from the Apiaceae family using multivariate analysis after LC-PDA-ESI-MS/MS and near infrared spectroscopy data are generated. Principal component analysis was successfully applied to distinguish between the nine Egyptian cultivars and the three German cultivars, and hierarchical cluster analysis also confirmed this distinctive clustering. Partial least square regression (PLS-R) models showed a relationship between phytochemicals and antioxidant activities. The metabolites responsible for the clustering pattern and variables important for projection (VIP) were identified, being twelve amongst nine Egyptian cultivar samples and thirteen amongst the Egyptian cultivar and the German cultivar comparison. The identified VIPs were also correlated with the antioxidant activity using PLS-R. In conclusion, the study showed novelty in the application of hyphenated analytical techniques and chemometrics that assist in quality control of herbal medicine.
Climate change will influence disease resistance breeding in wheat in Northwestern Europe
(2021) Miedaner, Thomas; Juroszek, Peter
Wheat productivity is threatened by global climate change. In several parts of NW Europe it will get warmer and dryer during the main crop growing period. The resulting likely lower realized on-farm crop yields must be kept by breeding for resistance against already existing and emerging diseases among other measures. Multi-disease resistance will get especially crucial. In this review, we focus on disease resistance breeding approaches in wheat, especially related to rust diseases and Fusarium head blight, because simulation studies of potential future disease risk have shown that these diseases will be increasingly relevant in the future. The long-term changes in disease occurrence must inevitably lead to adjustments of future resistance breeding strategies, whereby stability and durability of disease resistance under heat and water stress will be important in the future. In general, it would be important to focus on non-temperature sensitive resistance genes/QTLs. To conclude, research on the effects of heat and drought stress on disease resistance reactions must be given special attention in the future.
Comparative quantitative LC–MS/MS analysis of 13 amylase/trypsin inhibitors in ancient and modern Triticum species
(2020) Geisslitz, Sabrina; Longin, C. Friedrich H.; Koehler, Peter; Scherf, Katharina Anne
Amylase/trypsin inhibitors (ATIs) are major wheat allergens and they are also implicated in causing non-celiac gluten sensitivity and worsening other inflammatory conditions. With only few studies on ATI contents in different Triticum species available so far, we developed a targeted liquid chromatography-tandem mass spectrometry (LC–MS/MS) method based on stable isotope dilution assays to quantitate the 13 most important ATIs in a well-defined sample set of eight cultivars of common wheat and durum wheat (modern species), as well as spelt, emmer and einkorn (ancient species) grown at three locations in Germany, respectively. Only few ATIs with low contents were detected in einkorn. In contrast, spelt had the highest total ATI contents. Emmer and common wheat had similar total ATI contents, with durum wheat having lower contents than common wheat. Due to the lack of correlation, it was not possible to estimate ATI contents based on crude protein contents. The wheat species had a higher influence on ATI contents than the growing location and the heritability of this trait was high. Despite comparatively low intra-species variability, some cultivars were identified that may be promising candidates for breeding for naturally low ATI contents.
Comparison of aqueous and lactobacterial-fermented Mercurialis perennis L. (Dog’s Mercury) extracts with respect to their immunostimulating activity
(2023) Lorenz, Peter; Zilkowski, Ilona; Mailänder, Lilo K.; Klaiber, Iris; Nicolay, Sven; Garcia-Käufer, Manuel; Zimmermann-Klemd, Amy M.; Turek, Claudia; Stintzing, Florian C.; Kammerer, Dietmar R.; Gründemann, Carsten
Lactic acid (LA) fermentation of dog’s mercury (M. perennis L.) herbal parts was investigated in samples inoculated with either Lactobacteria (Lactobacillus plantarum and Pediococcus pentosaceus, LBF) or whey (WF). Depending on fermentation time, LA concentrations were monitored in a range of 3.4–15.6 g/L with a concomitant pH decline from 6.5 to 3.9. A broad spectrum of cinnamic acids depsides containing glucaric, malic and 2-hydroxyglutaric acids along with quercetin and kaempferol glycosides were detected by LC-DAD-ESI-MSn. Moreover, in this study novel constituents were also found both in unfermented and fermented extracts. Furthermore, amino acids and particular Lactobacteria metabolites such as biogenic amines (e.g., putrescine, 4-aminobutyric acid, cadaverine) and 5-oxoproline were assigned in WF extracts by GC-MS analysis after silylation. Enhanced NFκB and cytokine expression (IL-6, TNFα, IL-8 and IL-1β) was induced by all extracts, both non-fermented and fermented, in NFκB-THP-1 reporter cells, showing a concentration-dependent immunostimulatory effect. The WF extracts were tested for micronuclei formation in THP-1 cells and toxicity in luminescent bacteria (V. fischeri), whereby no mutagenic or toxic effects could be detected, which corroborates their safe use in pharmaceutical remedies.
Comparison of floral traits in Calibrachoa cultivars and assessment of their impacts on attractiveness to flower-visiting insects
(2021) Marquardt, Melanie; Kienbaum, Lydia; Losert, Dominik; Kretschmer, Lea Annina; Rigling, Marina; Zhang, Yanyan; Schweikert, Karsten; Westermann, Nils; Ruttensperger, Ute; Rosenkranz, Peter
Ornamental plants are appreciated by humans for their colorfulness, beauty, abundant flowering and long blooming periods. Many ornamental plants can also constitute an additional foraging resource for flower-visiting insects. However, the ability of the popular ornamental plant Calibrachoa to support urban insect communities is not well documented. In this study, 20 different Calibrachoa cultivars were selected and tested in regard to their insect friendliness based on standardized observations (I) in flight tents using the large earth bumble bee Bombus terrestris as a model species and (II) in open field trials. To investigate what floral characteristics might constitute attractiveness to bumble bees, various floral traits were recorded and compared across all tested Calibrachoa cultivars. Over a two-year period, a total of 6,327 foraging bumble bees were recorded in the tent observations. In the open field observations, we counted 4,188 flower-visiting insects. Our results revealed that (I) all Calibrachoa cultivars were visited by insects for foraging, (II) the number of insect visitors varied significantly among the 20 tested cultivars and (III) the cultivars displayed different floral traits. For the morphometric floral traits and the aroma profiles of Calibrachoa, only the mean nectar quantity and a few identified compounds could be correlated with attractiveness to the model species B. terrestris. We also found that the petal color of the tested cultivars had a significant impact on the number of visitors. Therefore, B. terrestris clearly preferred red or blue Calibrachoa cultivars over those with other petal colors. However, as the cultivar preferences in the different insect groups differed, it is highly recommended to use various cultivars in urban plantings. Nevertheless, efforts must be made to explain what additional floral traits make Calibrachoa and other ornamental plants generally attractive to flower visitors. This information can then be used for breeding purposes to increase the insect friendliness of ornamental plants.
Comparison of hybrid rye and wheat for grain yield and other agronomic traits under less favourable environmental conditions and two input levels
(2025) Miedaner, Thomas; Lauenstein, Sophie; Lieberherr, Bärbel; Tan, Daniel
Agriculture in the European Union is constantly the subject of public debate, particularly concerning reduced mineral fertilisation and reduced chemical plant protection. Hybrid rye could play a special role in this context, as it usually requires less fertiliser and pesticides than winter wheat and has a high yield potential. In Germany, both crops can be grown on most sites. To test whether hybrid rye is competitive with winter wheat, a trial was conducted with 10 hybrid rye and 20 wheat varieties of all quality levels. The trial was grown for three years (2021, 2022, 2023) on three conventionally managed farms with 40–64 soil points (on a scale of 1–100) in south-west Germany. It was conducted with two input levels: I1, fertilisation (100–120 kg N/ha for hybrid rye; 140–180 kg N/ha for winter wheat), and chemical plant protection including growth regulators; I2, no chemical plant protection, and 20% reduced N fertilisation, one to three harrows. In eight out of nine location × year combinations, hybrid rye was significantly ( p < 0.001) superior to winter wheat in terms of grain yield under these conditions. On average, hybrid rye yielded 9.1 and 7.8 t ha −1 for I1 and I2, respectively, compared to 8.0 and 6.8 t ha −1 for wheat, respectively. A close correlation of the cultivar’s grain yield between I1 and I2 for both crops showed a missing cultivar × input level interaction. Under these conditions, hybrid rye produced higher grain yields and better baking quality than wheat in both input regimes, contributing to a more sustainable cropping system.
Conserved worker policing in African carpenter ants with drastically different egg chemotypes
(2023) Buellesbach, Jan; Rinke, Janina; Reuter, Leonie; Thomas, Jurian M.; Hartmann, Vivien; Pohl, Marius; Gadau, Jürgen; Ernst, Ulrich R.
Distinctive chemical signatures have the potential to serve as discriminatory cues for olfactory recognition mechanisms. Cuticular hydrocarbon (CHC) profiles are among the most prominent chemical signatures in insects that can be highly diverse even among closely related species and between populations with similar ecology. Particularly within the major insect order Hymenoptera, CHC profiles are characterized by high complexity and variation with the potential to evolve rapidly. In this study, we found two very distinct CHC chemotypes distinguishing sympatric colonies of the African carpenter ant Camponotus maculatus (Hymenoptera: Formicinae). These chemotypic differences were mainly detected on the surface profiles of eggs produced by either queens or isolated worker groups. In one chemotype, queen- and worker-laid eggs are very similar. This is largely contrasted by the other chemotype, where queen-laid eggs clearly differ from worker-laid eggs with several prominent queen-exclusive compounds. However, workers display a stable behavior of discriminating against and selectively disposing of worker-laid eggs i.e., worker policing, independent of egg chemotype. Furthermore, genetic barcoding of workers revealed a clear separation between colonies characterized by producing these two distinct egg chemotypes, which may indicate that these colonies belong to a cryptic species complex. Interestingly, worker policing behaviour appears to be evolutionarily conserved, despite the strikingly different egg surface profiles.
Construction and description of a constitutive plipastatin mono-producing Bacillus subtilis
(2020) Vahidinasab, Maliheh; Lilge, Lars; Reinfurt, Aline; Pfannstiel, Jens; Henkel, Marius; Morabbi Heravi, Kambiz; Hausmann, Rudolf
Background: Plipastatin is a potent Bacillus antimicrobial lipopeptide with the prospect to replace conventional antifungal chemicals for controlling plant pathogens. However, the application of this lipopeptide has so far been investigated in a few cases, principally because of the yield in low concentration and unknown regulation of biosynthesis pathways. B. subtilis synthesizes plipastatin by a non-ribosomal peptide synthetase encoded by the ppsABCDE operon. In this study, B. subtilis 3NA (a non-sporulation strain) was engineered to gain more insights about plipastatin mono-production. Results: The 4-phosphopantetheinyl transferase Sfp posttranslationally converts non-ribosomal peptide synthetases from inactive apoforms into their active holoforms. In case of 3NA strain, sfp gene is inactive. Accordingly, the first step was an integration of a repaired sfp version in 3NA to construct strain BMV9. Subsequently, plipastatin production was doubled after integration of a fully expressed degQ version from B. subtilis DSM10T strain (strain BMV10), ensuring stimulation of DegU-P regulatory pathway that positively controls the ppsABSDE operon. Moreover, markerless substitution of the comparably weak native plipastatin promoter (Ppps) against the strong constitutive promoter Pveg led to approximately fivefold enhancement of plipastatin production in BMV11 compared to BMV9. Intriguingly, combination of both repaired degQ expression and promoter exchange (Ppps::Pveg) did not increase the plipastatin yield. Afterwards, deletion of surfactin (srfAA-AD) operon by the retaining the regulatory comS which is located within srfAB and is involved in natural competence development, resulted in the loss of plipastatin production in BMV9 and significantly decreased the plipastatin production of BMV11. We also observed that supplementation of ornithine as a precursor for plipastatin formation caused higher production of plipastatin in mono-producer strains, albeit with a modified pattern of plipastatin composition. Conclusions: This study provides evidence that degQ stimulates the native plipastatin production. Moreover, a full plipastatin production requires surfactin synthetase or some of its components. Furthermore, as another conclusion of this study, results point towards ornithine provision being an indispensable constituent for a plipastatin mono-producer B. subtilis strain. Therefore, targeting the ornithine metabolic flux might be a promising strategy to further investigate and enhance plipastatin production by B. subtilis plipastatin mono-producer strains.
Dairy byproducts as sustainable alternatives to FCS in 2D and 3D skeletal muscle cell cultures
(2025) Baldeweg, Tobias Horst; Hubel, Philipp; Günther, Johannes; Ostertag, Fabian; Nellinger, Svenja; Heine, Simon; Kluger, Petra Juliane
Skeletal muscle tissue engineering is a rapidly developing field with applications in disease modelling, tissue replacement, biorobotics, and cultivated meat. The need for more sustainable and ethical biotechnologies has grown due to concerns about resource scarcity, climate change, and animal welfare. One major challenge is replacing fetal calf serum (FCS), a widely used but ethically and environmentally highly problematic media supplement. A promising alternative is the utilization of natural byproducts such as whey and colostrum from the dairy industry, which provide essential nutrients and growth factors. In this study, wheys produced by microfiltration of raw milk and colostrum were investigated as FCS replacements for culturing C2C12 skeletal muscle cells. Composition analysis confirmed a variety of pro-proliferative compounds in both substances. Cell culture experiments led to the development of an optimized medium formulation based on colostrum whey. Colostrum whey medium (CM) supported cell proliferation and maintained the myogenic differentiation potential for over 30 days. Additionally, a CM-based freezing solution enabled effective cryopreservation throughout culture. In 3D static suspension culture, CM sustained viable spheroids for over 14 days. Spheroids showed significantly higher proliferation compared to those in serum-containing medium, making CM suitable for 3D modelling and scale-up of biomass production. These findings highlight CM as a sustainable, cost-effective, and ethical alternative for skeletal muscle tissue engineering, particularly in cultivated meat production.
Degradation of hop latent viroid during anaerobic digestion of infected hop harvest residues
(2021) Hagemann, Michael Helmut; Born, Ute; Sprich, Elke; Seigner, Luitgardis; Oechsner, Hans; Hülsemann, Benedikt; Steinbrenner, Jörg; Winterhagen, Patrick; Lehmair, Erich
The citrus bark cracking viroid (CBCVd) was identified as causal agent for a severe stunting disease in hops. Viroids are highly stable parasitic RNAs, which can be easily transmitted by agricultural practices. Since CBCVd has recently been detected in two European countries a growing concern is that this pathogen will further spread and thereby threaten the European hop production. Biogas fermentation is used to sanitize hop harvest residues infected with pathogenic fungi. Consequently, the aim of this study was to test if biogas fermentation can contribute to viroid degradation at mesophilic (40 °C) and thermophilic (50 °C) conditions. Therefore, a duplex reverse transcription real-time PCR analysis was developed for CBCVd and HLVd detection in biogas fermentation residues. The non-pathogenic hop latent viroid (HLVd) was used as viroid model for the pathogenic CBCVd. The fermentation trials showed that HLVd was significantly degraded after 30 days at mesophilic or after 5 days at thermophilic conditions, respectively. However, sequencing revealed that HLVd was not fully degraded even after 90 days. The incubation of hop harvest residues at different temperatures between 20 and 70 °C showed that 70 °C led to a significant HLVd degradation after 1 day. In conclusion, we suggest combining 70 °C pretreatment and thermophilic fermentation for efficient viroid decontamination.
Detecting multiple structural breaks in systems of linear regression equations with integrated and stationary regressors
(2025) Schweikert, Karsten
In this paper, we propose a two‐step procedure based on the group LASSO estimator in combination with a backward elimination algorithm to detect multiple structural breaks in linear regressions with multivariate responses. Applying the two‐step estimator, we jointly detect the number and location of structural breaks and provide consistent estimates of the coefficients. Our framework is flexible enough to allow for a mix of integrated and stationary regressors, as well as deterministic terms. Using simulation experiments, we show that the proposed two‐step estimator performs competitively against the likelihood‐based approach in finite samples. However, the two‐step estimator is computationally much more efficient. An economic application to the identification of structural breaks in the term structure of interest rates illustrates this methodology.

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