Biology subjects

Benz, J. P.

Publications and source records attributed to Benz, J. P..

12 recordsLinked to original sources

bioRxiv2025

Tree bark as a substrate for mycelium-bound composites with two Ganoderma species

Bark is currently considered as a by-product of the wood industry and is mostly incinerated for energy, left in forests, or used as a mulch layer in gardens, parks, and forests to help prevent soil from drying out. However, considering that bark makes up about 10-20% of the tree volume, there is a considerable amount of material that can be exploited and should be investigated in terms of a resource-efficient bio-economy. One way to use bark as a raw material for innovative products could be as a substrate within mycelium-bound composite materials. However, since one of the natural functions of bark is to inhibit microbial infestation of the trees, it was unclear whether bark could be utilized in this manner. Therefore, we investigate in this study the possibilities of producing such composites by evaluating the performance of several bark-fungus combinations. Three different barks (from Douglas fir, Scots pine and European birch) and two species of fungi (Ganoderma sessile and Ganoderma adspersum) were selected for the experiments. Mycelium growth rates were evaluated with a newly developed method using fungal "growth tubes". In addition, composites were prepared for performance tests from pure bark and 1:1 mixtures of bark and beech wood sawdust. Composites made of mixed bark and beech wood were mostly well overgrown with a thick layer of mycelium on the surface, supporting higher compression strengths. The mycelium layer on the composites made with tree bark only was considerably thinner, resulting in lower compression strength. Water absorption potential was found to be highly dependent on the mycelium layer on the composite surfaces, which has substantial hydrophobic properties. Overall, although the required incubation times tend to be longer than for other commonly used substrates, our experiments demonstrate that bark clearly represents a potential co-substrate for the production of mycelium-bound composites. Nevertheless, further studies with different fungi and barks are required to identify mixtures for specific applications that require shorter incubation times.

bioengineering↗

bioRxiv2025

Improving Seedling Survival for Forest Restorations: A Novel Screening Method to Identify Microbial Allies Against Drought Stress

Improving drought tolerance of tree seedlings by plant growth-promoting microorganisms (PGPMs) is a promising approach for nature-based forest restoration. Identifying suitable microorganisms requires a robust selection, including efficient in planta screenings. We sampled at two forest sites in southern Germany with drought legacies and within a dry period to enhance the probability of isolating drought-tolerant microbes. Metabarcoding of the resident soil community revealed a broad on-site diversity with the potential for diverse plant growth-promoting and stress-resistance traits. We isolated 1,292 bacteria and 59 fungi from fine roots of Norway spruce and European beech. 429 isolates were identified on genus level. The most abundant genera were Paraburkholderia (121) and Bacillus (43) in bacteria and Penicillium (8) and Umbelopsis (8) in fungi. Isolates were scored in vitro for abiotic stress tolerance and plant growth-promoting traits, revealing diverse plant growth-promoting abilities for 31 bacteria and a particularly high stress tolerance for 8 fungi. Importantly, an axenic 24-well-plate system was developed to test the most promising bacteria on spruce seedlings under drought. The system allowed direct comparison of inoculation effects on seedling growth and survival with or without drought application. Two strains improved survival and root length under drought, while six strains significantly promoted plant growth under well-watered conditions. This study represents one of the first larger scale screenings for PGPMs isolated from forest soils on tree seedlings under drought and may contribute to finding nature-based drought mitigation strategies.

microbiology↗

bioRxiv2024

Population genomics of Macrophomina spp. reveals cryptic host specialization and evidence for meiotic recombination

Knowledge of the factors structuring populations of pathogenic fungi is fundamental to disease management efforts and basic biology. High-quality short-read sequence data were obtained for 463 Macrophomina spp. isolates collected from 91 host plant species and soil in 23 countries. Analyses revealed high diversity, admixture, and equal mating type ratios suggesting on-going recombination. Although most tested isolates could asymptomatically colonize strawberry, only isolates from a single phylogroup caused disease. In addition to strawberry, evidence for host specialization was discovered for soybean, demonstrating this broad host range pathogen contains phylogroups with cryptic specialization. Geography x isolate genotype associations were weak, suggesting these species were frequently trafficked between regions. Re-analysis using genomic data supported current species boundaries, and new molecular markers were designed to specifically identify each species. Contrary to expectations, M. phaseolina should be considered a species with both specialist and generalist populations for which meiosis can increase genetic diversity.

genomics↗

bioRxiv2024

Deciphering plant-beneficial fungal interactions: Unravelling metabolic diversity that underpins communication between Laccaria bicolor and Trichoderma

With over 250 known species, the genus Trichoderma (Ascomycota, Hypocreaceae) is found in various soils, on plant surfaces and as plant endophytes. While Trichoderma species are known as mycoparasites, their antagonistic behaviour can also negatively affect other beneficial fungi, such as mycorrhizal fungi. To gain insight into the metabolic signals involved in the interactions between the ectomycorrhizal fungus (ECM) Laccaria bicolor (Basidomycota, Hydnangiaceae), and different mycoparasitic Trichoderma spp. (T. harzianum strains WM24a1, MS8a1 and ES8g1, and T. atrobrunneum), we performed in vitro dual-confrontation experiments. We studied the volatile organic compounds (VOCs), hyphal metabolomes and soluble metabolites released by each of the fungi in various co-cultivation scenarios. The results revealed an altered growth of the mycelia depending on the degree of contact: When Trichoderma spp. and L. bicolor shared only the same headspace, Trichoderma spp. growth was at least partially inhibited, whereas in direct contact the growth of L. bicolor was impaired. Distinct strain- and species-specific changes in hyphal metabolites, in exudates and volatile emission were revealed from each of the studied fungi. We identified both core metabolite profiles and interaction-specific metabolic responses that were related to carbohydrate, lipid, nucleotide, energy and amino acid metabolisms. Volatile and soluble metabolites revealed temporal and spatial adjustments in dual cultures compared to solitary cultures, suggesting rapid contact-dependent adaptations and demonstrating the dynamic communication mechanisms between Trichoderma spp. and the ECM. These results suggest a central role for both emitted and secreted fungal metabolites in the fungal non-self-recognition and in interaction with each other.

microbiology↗

bioRxiv2024

Lifestyle-specific responses of Trichoderma spp. in mycoparasitic confrontations and implications for biocontrol of Populus x canescens

O_LITrichoderma spp. are gaining popularity in agriculture and forestry due to their multifaceted roles in promoting plant growth through e.g. nutrient translocation, hormone production, induction of plant systemic resistance, but also direct antagonism of other fungi. However, the mycotrophic nature of the genus bears the risk of possible interference with other native plant-beneficial fungi, such as ectomycorrhiza, in the rhizosphere. Such interference could yield unpredictable consequences for the host plants of these ecosystems. C_LIO_LIWe investigated whether Trichoderma spp. can differentiate between beneficial ectomycorrhizal fungi (represented by Laccaria bicolor and Hebeloma cylindrosporum) and pathogenic fungi (represented by Fusarium graminearum and Alternaria alternata) in different confrontation scenarios, including a newly developed olfactometer "race tube"-like system. C_LIO_LIUsing two independent species, T. harzianum, and T. atrobrunneum, with plant-growth-promoting and immune-stimulating properties towards Populus x canescens, our study revealed robustly accelerated growth towards phytopathogens, while showing a contrary response to ectomycorrhizal fungi. Transcriptomic analyses identified distinct genetic programs during interaction corresponding to the lifestyles, emphasizing the expression of mycoparasitism-related genes only in the presence of phytopathogens. C_LIO_LIThe findings reveal a critical mode of fungal community interactions belowground and suggest that Trichoderma spp. can distinguish between fungal partners of different lifestyles already at a distance. C_LI

microbiology↗

bioRxiv2022

Characterization of the Rhizophagus irregularis multicopper oxidase family indicates that the iron transporter RiFTR1 does not require a ferroxidase partner

The contribution of arbuscular mycorrhizal fungi (AM fungi) to plant iron (Fe) acquisition has been demonstrated in several studies. Recently, it has been shown that AM fungi use a high-affinity reductive pathway for Fe uptake. In the AM fungus Rhizophagus irregularis the ferric reductase RiFRE1 and the Fe permeases RiFTR1 and RiFTR2 have already been characterized. In an attempt to identify the third component of the reductive iron uptake pathway, a genome-wide approach has been used in R. irregularis to find genes encoding ferroxidases of the multicopper oxidase (MCO) gene family. Nine genes putatively encoding MCOs (RiMCO1-9) were identified. A phylogenetic analysis of MCO sequences of fungi from different taxonomic groups revealed that all RiMCOs clustered together in the ferroxidase/laccase group, and none with the Fet3-type ferroxidases. RiMCO1 and RiMCO3 were the only MCO genes displaying a detectable gene expression pattern typical of a high-affinity Fe transport system, indicating that RiMCO1 and RiMCO3 might have a role in the reductive high-affinity Fe uptake system. Moreover, yeast mutant complementation assays showed that the iron permease RiFTR1 can operate without the presence of a ferroxidase, indicating that it is able to transport also ferrous (II) iron.

molecular biology↗

bioRxiv2019

Comparative evaluation of Aspergillus niger strains for endogenous pectin depolymerization capacity and suitability for D-galacturonic acid production

Pectinaceous agricultural residues rich in O_SCPLOWDC_SCPLOW-galacturonic acid (O_SCPLOWDC_SCPLOW-GalA), such as sugar beet pulp, are considered as promising feedstocks for waste-to-value conversions. Aspergillus niger is known for its strong pectinolytic activity. However, while specialized strains for production of citric acid or proteins are openly available, this is not the case for the production of pectinases. We therefore systematically compared the pectinolytic capabilities of six A. niger strains (ATCC 1015, ATCC 11414, NRRL 3122, CBS 513.88, NRRL 3, N402) using controlled batch cultivations in stirred-tank bioreactors. A. niger ATCC 11414 showed the highest polygalacturonase activity, specific protein secretion and a suitable morphology. Furthermore, O_SCPLOWDC_SCPLOW-GalA release from sugar beet pulp was 75% higher compared to the standard lab strain A. niger N402. Our study therefore presents a robust initial strain selection to guide future process improvement of O_SCPLOWDC_SCPLOW-GalA production from agricultural residues and identifies the most suitable base strain for further genetic optimizations.

bioengineering↗

bioRxiv2019

Broad substrate-specific phosphorylation events are associated with the initial stage of plant cell wall recognition in Neurospora crassa

Fungal plant cell wall degradation processes are governed by complex regulatory mechanisms, allowing the organisms to adapt their metabolic program with high specificity to the available substrates. While the uptake of representative plant cell wall mono- and disaccharides is known to induce specific transcriptional and translational responses, the processes related to early signal reception and transduction remain largely unkown. A fast and reversible way of signal transmission are post-translational protein modifications, such as phosphorylations, which could initiate rapid adaptations of the fungal metabolism to a new condition. To elucidate how changes in the initial substrate recognition phase of Neurospora crassa affect the global phosphorylation pattern, phospho-proteomics was performed after a short (2 minutes) induction period with several plant cell wall-related mono- and disaccharides. The MS/MS-based peptide analysis revealed large-scale substrate-specific protein phosphorylation and de-phosphorylations. Using the proteins identified by MS/MS, a protein-protein-interaction (PPI) network was constructed. The variance in phosphorylation of a large number of kinases, phosphatases and transcription factors indicate the participation of many known signaling pathways, including circadian responses, two-component regulatory systems, MAP kinases as well as the cAMP-dependent and heterotrimeric G-protein pathways. Adenylate cyclase, a key component of the cAMP pathway, was identified as a potential hub for carbon source-specific differential protein interactions. In addition, four phosphorylated F-Box proteins were identified, two of which, Fbx-19 and Fbx-22, were found to be involved in carbon catabolite repression responses. Overall, these results provide unprecedented and detailed insights into a so far less well known stage of the fungal response to environmental cues and allow to better elucidate the molecular mechanisms of sensory perception and signal transduction during plant cell wall degradation.

molecular biology↗

bioRxiv2019

Preserving cultural heritage: Analyzing the antifungal potential of ionic liquids tested in paper restoration

Early industrialization and the development of cheap production processes for paper have led to an exponential accumulation of paper-based documents during the last two centuries. Archives and libraries harbor vast amounts of ancient and modern documents and have to undertake extensive endeavors to protect them from abiotic and biotic deterioration. While services for mechanical preservation such as ex post de-acidification of historic documents are already commercially available, the possibilities for long-term protection of paper-based documents against fungal attack (apart from temperature and humidity control) are very limited. Novel processes for mechanical enhancement of damaged cellulosic documents use Ionic Liquids (IL) as essential process components. With some of these ILs having azole-functionalities similar to well-known fungicides such as Clotrimazole, the possibility of antifungal activities of these ILs was proposed but has not yet been experimentally confirmed.\n\nWe evaluated the potency of four ILs with potential application in paper restoration for suppression of fungal growth on five relevant paper-infesting molds. The results revealed a general antifungal activity of all ILs, which increased with the size of the non-polar group. Physiological experiments and ultimate elemental analysis allowed to determine the minimal inhibitory concentration of each IL as well as the residual IL concentration in process-treated paper. These results provide valuable guidelines for IL-applications in paper restoration processes with antifungal activity as an added benefit. With azoles remaining in the paper after the process, simultaneous repair and biotic protection in treated documents could be facilitated.

microbiology↗

bioRxiv2019

Genome-wide and enzymatic analysis reveals efficient D-galacturonic acid metabolism in the basidiomycete yeast Rhodosporidium toruloides

Biorefining of renewable feedstocks is one of the most promising routes to replace fossil-based products. Since many common fermentation hosts, such as Saccharomyces cerevisiae, are naturally unable to convert many component plant cell wall polysaccharides, the identification of organisms with broad catabolism capabilities represents an opportunity to expand the range of substrates used in fermentation biorefinery approaches. The red basidiomycete yeast Rhodosporidium toruloides is a promising and robust host for lipid and terpene derived chemicals. Previous studies demonstrated assimilation of a range of substrates, from C5/C6-sugars to aromatic molecules similar to lignin monomers. In the current study, we analyzed R. toruloides potential to assimilate D-galacturonic acid, a major sugar in many pectin-rich agricultural waste streams, including sugar beet pulp and citrus peels. D-galacturonic acid is not a preferred substrate for many fungi, but its metabolism was found to be on par with D-glucose and D-xylose in R. toruloides. A genome-wide analysis by combined RNAseq/RB-TDNAseq revealed those genes with high relevance for fitness on D-galacturonic acid. While R. toruloides was found to utilize the same non-phosphorylative catabolic pathway known from ascomycetes, the maximal velocities of several enzymes exceeded those previously reported. In addition, an efficient downstream glycerol catabolism and a novel transcription factor were found to be important for D-galacturonic acid utilization. These results set the basis for use of R. toruloides as a potential host for pectin-rich waste conversions and demonstrate its suitability as a model for metabolic studies in basidiomycetes.\n\nImportanceThe switch from the traditional fossil-based industry to a green and sustainable bio-economy demands the complete utilization of renewable feedstocks. Many currently used bio-conversion hosts are unable to utilize major components of plant biomass, warranting the identification of microorganisms with broader catabolic capacity and characterization of their unique biochemical pathways. D-galacturonic acid is a plant component of bio-conversion interest and is the major backbone sugar of pectin, a plant cell wall polysaccharide abundant in soft and young plant tissues. The red basidiomycete and oleaginous yeast Rhodosporidium toruloides has been previously shown to utilize a range of sugars and aromatic molecules. Using state-of-the-art functional genomic methods, physiological and biochemical assays, we elucidated the molecular basis underlying the efficient metabolism of D-galacturonic acid. This study identifies an efficient pathway for uronic acid conversion to guide future engineering efforts, and represents the first detailed metabolic analysis of pectin metabolism in a basidiomycete fungus.

systems biology↗

bioRxiv2019

Cross-talk of cellulose and mannan perception pathways leads to inhibition of cellulase production in several filamentous fungi

It is essential for microbes to acquire information about their environment. Fungi use soluble degradation products of plant cell wall components to understand the substrate composition they grow on. Individual signaling pathways have been well described. However, the interconnections between pathways remain poorly understood. In the present work, we provide evidence of \"confusion\" due to cross-talk between the perception pathways for cellulose and the hemicellulose mannan in several filamentous fungi, leading to the inhibition of cellulase expression. We used the functional genomics tools available for Neurospora crassa to investigate this signaling overlap at the molecular level. Cross-talk and competitive inhibition could be identified both during uptake by cellodextrin transporters and intracellularly. Importantly, the overlap is independent of CRE-1-mediated catabolite repression. These results provide novel insights into the regulatory networks of lignocellulolytic fungi and will contribute to the rational optimization of fungal enzyme production for efficient plant biomass depolymerization and utilization.

genetics↗

bioRxiv2017

Network Reconstruction And Systems Analysis Of Plant Cell Wall Deconstruction By Neurospora crassa

Plant biomass degradation by fungal derived enzymes is rapidly expanding in economic importance as a clean and efficient source for biofuels. The ability to rationally engineer filamentous fungi would facilitate biotechnological applications for degradation of plant cell wall polysaccharides. However, incomplete knowledge of biomolecular networks responsible for plant cell wall deconstruction impedes experimental efforts in this direction. To expand this knowledge base, a detailed network of reactions important for deconstruction of plant cell wall polysaccharides into simple sugars was constructed for the filamentous fungus Neurospora crassa. To reconstruct this network, information was integrated from five heterogeneous data types: functional genomics, transcriptomics, proteomics, genetics, and biochemical characterizations. The combined information was encapsulated into a feature matrix and the evidence weighed to assign annotation confidence scores for each gene within the network. Comparative analyses of RNA-seq and ChIP-seq data shed light on the regulation of the plant cell wall degradation network (PCWDN), leading to a novel hypothesis for degradation of the hemicellulose mannan. The transcription factor CLR-2 was subsequently experimentally shown to play a key role in the mannan degradation pathway of Neurospora crassa. Our network serves as a scaffold for integration of diverse experimental data, leading to elucidation of regulatory design principles for plant cell wall deconstruction by filamentous fungi, and guiding efforts to rationally engineer industrially relevant hyper-production strains.

systems biology↗