Biology subjects

Jacobsen, C. S.

Publications and source records attributed to Jacobsen, C. S..

6 recordsLinked to original sources

bioRxiv2026

TriMic: a Triticum aestivum microbial culture collection and synthetic community for dissecting wheat-microbe interactions

Understanding the molecular mechanisms underlying plant-microbe interactions is essential for developing innovative microbe-based agrotechnologies. However, deciphering these mechanisms within the complexity of natural microbial communities remains challenging. Such challenges can be addressed by employing synthetic microbial communities (SynComs) derived from well characterized microbial culture collections. Despite their importance, plant-associated microbial collections from major agricultural crops remain scarce. To bridge this gap, we established TriMic, a taxonomically and functionally representative culture collection of wheat root-associated bacteria. Complementing this collection, we include high quality genome sequences and an overview of genes involved in plant colonization, nutrient cycling, and plant growth promotion. Furthermore, we expanded this experimental toolkit by designing a reduced complexity SynCom that enables controlled dissection of plant-microbe interactions. Together, these resources lay the groundwork for mechanistic studies of plant-microbe interactions to accelerate biostimulant development aimed at enhancing agricultural productivity and sustainability. The TriMic collection and whole genomes are publicly available at the DSMZ (https://www.dsmz.de/collection/catalogue/microorganisms/microbiota/trimic) and NCBI. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=166 SRC="FIGDIR/small/740170v1_ufig1.gif" ALT="Figure 1"> View larger version (54K): org.highwire.dtl.DTLVardef@91bda0org.highwire.dtl.DTLVardef@386c49org.highwire.dtl.DTLVardef@4d455aorg.highwire.dtl.DTLVardef@1043ccb_HPS_FORMAT_FIGEXP M_FIG C_FIG Overview of the workflow for development of the wheat root bacterial collection and SynCom. (a) Bacterial isolates were recovered from roots of wheat grown in natural soil using two strategies after a root slurry was prepared: directly plating or host-mediated, which involved producing a secondary root slurry from wheat roots inoculated with the original root slurry (Ben Niu et al. 2017). Both root slurries were diluted and then plated onto four different types of media (1/10 R2A, 1/20 R2A, 1/100 R2A, and VxylG). Colonies were picked based on morphology and time of appearance. (b) Non-redundant strains of 88 purified isolates were made publicly available at the German Collection of Microorganisms and Cell cultures GmbH (DSMZ). (c) A 15-species synthetic community (SynCom) was developed after evaluating community dynamics in wheat roots inoculated with a 27-member consortium selected across genera in the collection. (d) Isolate genomes were sequenced using either short-read or hybrid assemblies with long-reads. Following functional annotation, genomes were screened for traits associated with plant-microbe interactions and secondary metabolite production.

microbiology↗

bioRxiv2023

Concurrent stimulation of diflufenican biodegradation and changes in the active microbiome in gravel revealed by Total RNA

The use of slowly degrading pesticides poses a particular problem when these are applied to urban areas such as gravel paths. The urban gravel provides an environment very different from agricultural soils; i.e., it is both lower in carbon and microbial activity. We, therefore, endeavoured to stimulate the degradation of the pesticide diflufenican added to an urban gravel microcosm amended with dry alfalfa to increase microbial activity. In the present study, the formation of the primary diflufenican metabolite 2-[3-(Trifluoromethyl)phenoxy]nicotinic acid (commonly abbreviated as AE-B) was stimulated by the alfalfa amendment. The concurrent changes of the active microbial communities within the gravel were explored using shotgun metatranscriptomic sequencing of ribosomal RNA and messenger RNA. Our results showed, that while the active microbial communities in the gravel were dominated by bacteria with a relative abundance of 87.0 - 98.5 %, the eukaryotic groups, fungi and micro-eukaryotes, both had a 4-5 fold increase in relative abundance over time in the alfalfa amended treatment. Specifically, the relative abundance of microorganisms involved in degradation of complex carbon sources, Bacteroidetes, Verrucomicrobia, Sordariomycetes, Mortierellales, and Tremellales, were shown to increase in the alfalfa amended treatment. Further, the functional gene profile showed an increase in genes involved in increased activity and production of new biomass in the alfalfa treatment compared to the control, as well as pointing to genes potentially involved in biodegradation of complex carbon sources and the biotransformation of diflufenican.

microbiology↗

bioRxiv2022

Abrupt permafrost thaw triggers microbial bloom and increased activity of microbial predators

Permafrost soils store a substantial part of the global soil carbon and nitrogen. However global warming causes abrupt erosion and gradual thaw, which make these stocks vulnerable to microbial decomposition into greenhouse gases. Here, we investigated the microbial response to abrupt in situ permafrost thaw. We sequenced the total RNA of a 1 m deep soil core consisting of up to 26500-year-old permafrost material from an active abrupt erosion site. We analysed the microbial community in the active layer soil, the recently thawed, and the intact permafrost and found maximum RNA:DNA ratios indicating a microbial bloom in recently thawed permafrost. Several fast-growing prokaryotic taxa dominated thawed permafrost, including Sphingobacteriales, Burkholderiales, and Nitrosomonadales. Overall, the thaw state and soil moisture consistently explained changes in community composition, with especially the permafrost community being significantly distinct from thawed soils. Predation correlated with changes in prokaryotic composition. Bacterial grazers were dominated by Myxococcales and abundant in the active layer. In contrast, protozoa, especially Cercozoa and Ciliophora, doubled in relative abundance in thawed layers. Our findings highlight the ecological importance of a rapid development of microbial blooms as well as the successive predation as biological control mechanism in abruptly thawing permafrost. One sentence summaryUsing total RNA from an up to 26500-year-old abruptly eroding permafrost site in Greenland, we described a microbial bloom and its controls, including bacterial and microeukaryotic predators. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=146 SRC="FIGDIR/small/499897v2_ufig1.gif" ALT="Figure 1"> View larger version (51K): org.highwire.dtl.DTLVardef@10ab79eorg.highwire.dtl.DTLVardef@9d0146org.highwire.dtl.DTLVardef@1bec509org.highwire.dtl.DTLVardef@179f397_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2019

To assemble or not to resemble -- A validated Comparative Metatranscriptomics Workflow (CoMW)

BackgroundMetatranscriptomics has been used widely for investigation and quantification of microbial communities activity in response to external stimuli. By assessing the genes expressed, metatranscriptomics provide an understanding of the interactions between different major functional guilds and the environment. Here, we present de-novo assembly-based Comparative Metatranscriptomics Workflow (CoMW) implemented in a modular, reproducible structure, significantly improving the annotation and quantification of metatranscriptomes. Metatranscriptomics typically utilize short sequence reads, which can either be directly aligned to external reference databases (\"assembly-free approach\") or first assembled into contigs before alignment (\"assembly-based approach\"). We also compare CoMW (assembly-based implementation) with assembly-free alternative workflow, using simulated and real-world metatranscriptomes from Arctic and Temperate terrestrial environments. We evaluate their accuracy in precision and recall using generic and specialized hierarchical protein databases.\n\nResultsCoMW provided significantly fewer false positives resulting in more precise identification and quantification of functional genes in metatranscriptomes. Using the comprehensive database M5nr, the assembly-based approach identified genes with only 0.6% false positives at thresholds ranging from inclusive to stringent compared to the assembly-free approach yielding up to 15% false positives. Using specialized databases (Carbohydrate Active-enzyme and Nitrogen Cycle), the assembly-based approach identified and quantified genes with 3-5x less false positives. We also evaluated the impact of both approaches on real-world datasets.\n\nConclusionsWe present an open source de-novo assembly-based Comparative Metatranscriptomics Workflow (CoMW). Our benchmarking findings support the argument of assembling short reads into contigs before alignment to a reference database, since this provides higher precision and minimizes false positives.

bioinformatics↗

bioRxiv2019

Transcriptomic responses to warming and cooling of an Arctic tundra soil microbiome

BackgroundArctic surface soils experience pronounced seasonal changes in temperature and chemistry. However, it is unclear how these changes affect microbial degradation of organic matter, nitrogen cycling and microbial stress responses. We combined measurements of microbiome transcriptional activity, CO2 production, and pools of carbon and nitrogen to investigate the microbial response to warming in the laboratory, from -10 {degrees}C to 2 {degrees}C, and subsequent cooling, from 2 {degrees}C to -10 {degrees}C, of a high Arctic tundra soil from Svalbard, Norway.\n\nResultsGene expression was unaffected by warming from -10 {degrees}C to -2 {degrees}C and by cooling from -2 {degrees}C to -10 {degrees}C, while upon freezing (2 {degrees}C to -2 {degrees}C) a defense response against oxidative stress was observed. Following modest transcriptional changes one day after soil thaw, a more pronounced response was observed after 17 days, involving numerous functions dominated by an upregulation of genes involved in transcription, translation and chaperone activity. Transcripts related to carbohydrate metabolism and degradation of complex polymers (e.g. cellulose, hemicellulose and chitin) were also enhanced following 17 days of soil thaw, which was accompanied by a four-fold increase in CO2 production. In addition, anaerobic ammonium oxidation and turnover of organic nitrogen were upregulated. In contrast, nitrification, denitrification and assimilatory nitrate reduction were downregulated leading to an increase in the concentration of soil inorganic nitrogen.\n\nConclusionthe microorganisms showed negligible response to changes in sub-zero temperatures and a delayed response to thaw, which after 17 days led to upregulation of soil organic matter degradation and enhanced CO2 production, as well as downregulation of key pathways in nitrogen cycling and a concomitant accumulation of inorganic nitrogen available for plants.

microbiology↗

bioRxiv2018

Quantitative and qualitative evaluation of the impact of the G2 enhancer, bead sizes and lysing tubes on the bacterial community composition during DNA extraction from recalcitrant soil core samples based on community sequencing and qPCR

Soil DNA extraction encounters numerous challenges that can affect both yield and purity of the recovered DNA. Clay particles lead to reduced DNA extraction efficiency, and PCR inhibitors from the soil matrix can negatively affect downstream analyses when applying DNA sequencing. Further, these effects impede molecular analysis of bacterial community compositions in lower biomass samples, as often observed in deeper soil layers. Many studies avoid these complications by using indirect DNA extraction with prior separation of the cells from the matrix, but such methods introduce other biases that influence the resulting microbial community composition.\n\nTo address these issues, a direct DNA extraction method was applied in combination with the use of a commercial product, the G2 DNA/RNA Enhancer(R), marketed as being capable of improving the amount of DNA recovered after the lysis step. The results showed that application of G2 increased DNA yields from the studied clayey soils from layers between 1.00 and 2.20 m below ground level.\n\nImportantly, the use of G2 did not introduce bias, as it did not result in any significant differences in the biodiversity of the bacterial community measured in terms of alpha and beta diversity and taxonomical composition.\n\nFinally, this study considered a set of customised lysing tubes for evaluating possible influences on the DNA yield. Tubes customization included different bead sizes and amounts, along with lysing tubes coming from two suppliers. Results showed that the lysing tubes with mixed beads allowed greater DNA recovery compared to the use of either 0.1 or 1.4 mm beads, irrespective of the tube supplier.\n\nThese outcomes may help to improve commercial products in DNA/RNA extraction kits, besides raising awareness about the optimal choice of additives, offering opportunities for acquiring a better understanding of topics such as vertical microbial characterisation and environmental DNA recovery in low biomass samples.

microbiology↗