Einfluss von Bewirtschaftungsmaßnahmen auf die Struktur und Funktion der Bodenmikroflora

Im ökologischen Landbau gilt es unter umweltschonender Bewirtschaftung trotzdem hohe Erträge zu erzielen. Bisher wurde dies durch konventionelle Pflügung realisiert, diese Maßnahme zerstört aber die Bodenstruktur und damit den Lebensraum wichtiger Organismengruppen. Daher ist es wünschenswert die im konventionellen Landbau schon übliche reduzierte Bodenbearbeitung auch im ökologischen Landbau zu etablieren. Im Rahmen dieses Projektes wurde daher untersucht wie sich die reduzierte Bodenbearbeitung auf das Mikrobiom auswirkt. Da Pflügen insbesondere die Bodenstruktur zerstört, lag der besondere Fokus auf dem bakteriellen Potential strukturbildende Substanzen wie Exo-(EPS) und Lipopolysaccharide (LPS) zu produzieren. Dazu wurden vier Standorte mit unterschiedlicher Bodentextur untersucht. Das Potential zur Bildung von EPS/LPS war generell im Pflughorizont am größten. Als Indikatorgene wurden wza für die EPS-Synthese und lptG und lptF für den LPS Transport identifiziert. Während die Abundanz der Gene nicht durch die Bodenbearbeitung beeinflusst wurde, hat sich die Zusammensetzung der Schlüsselorganismen je nach Standort, Tiefe und Bodenbearbeitung unterschieden. Da die strukturbildenden Eigenschaften der Polysaccharide bei jedem Organismus anders sind, können kleine Unterschiede in der mikrobiellen Zusammensetzung zu großen Unterschieden in der Aggregatstabilität führen. So hat sich gezeigt, dass trotz vergleichbarer relativer Genabundanzen schluffiger Boden am sensibelsten auf die Bodenbearbeitung reagiert, mit höheren Werten unter reduzierter Bodenbearbeitung. Die sowieso schon schlechte Aggregierung in sandigen Böden, konnte nicht verbessert werden. Alternativ könnte man hier aber durch reduzierte Bodenbearbeitung die Entwicklung von BBK unterstützen, die wie sich gezeigt hat ein großes Potential für die Speicherung von Nährstoffen haben, als auch die Bildung von EPS/LPS. Interessanterweise haben die Netzwerkanalysen ergeben, dass insbesondere Bakterien, die das Pflanzenwachstum unterstützen wie Micromonapsora und Actinobacteria durch die Bodenbearbeitung beeinflusst werden. Sie akkumulieren im Oberboden bei reduzierter Bearbeitung und unter dem Pflughorizont bei den Pflugvarianten und folgen somit der Wurzelpenetrationstiefe

Functional Traits Co-Occurring with Mobile Genetic Elements in the Microbiome of the Atacama Desert

Mobile genetic elements (MGEs) play an essential role in bacterial adaptation and evolution. These elements are enriched within bacterial communities from extreme environments. However, very little is known if specific genes co-occur with MGEs in extreme environments and, if so, what their function is. We used shotgun-sequencing to analyse the metagenomes of 12 soil samples and characterized the composition of MGEs and the genes co-occurring with them. The samples ranged from less arid coastal sites to the inland hyperarid core of the Atacama Desert, as well as from sediments below boulders, protected from UV-irradiation. MGEs were enriched at the hyperarid sites compared with sediments from below boulders and less arid sites. MGEs were mostly co-occurring with genes belonging to the Cluster Orthologous Group (COG) categories “replication, recombination and repair,” “transcription” and “signal transduction mechanisms.” In general, genes coding for transcriptional regulators and histidine kinases were the most abundant genes proximal to MGEs. Genes involved in energy production were significantly enriched close to MGEs at the hyperarid sites. For example, dehydrogenases, reductases, hydrolases and chlorite dismutase and other enzymes linked to nitrogen metabolism such as nitrite- and nitro-reductase. Stress response genes, including genes involved in antimicrobial and heavy metal resistance genes, were rarely found near MGEs. The present study suggests that MGEs could play an essential role in the adaptation of the soil microbiome in hyperarid desert soils by the modulation of housekeeping genes such as those involved in energy production.EC/FP7/339231/EU/Habitability of Martian Environments: Exploring the Physiological and Environmental Limits of Life/HOM

The Influence of Land Use Intensity on the Plant-Associated Microbiome of Dactylis glomerata L.

In this study, we investigated the impact of different land use intensities (LUI) on the root-associated microbiome of Dactylis glomerata (orchardgrass). For this purpose, eight sampling sites with different land use intensity levels but comparable soil properties were selected in the southwest of Germany. Experimental plots covered land use levels from natural grassland up to intensively managed meadows. We used 16S rRNA gene based barcoding to assess the plant-associated community structure in the endosphere, rhizosphere and bulk soil of D. glomerata. Samples were taken at the reproductive stage of the plant in early summer. Our data indicated that roots harbor a distinct bacterial community, which clearly differed from the microbiome of the rhizosphere and bulk soil. Our results revealed Pseudomonadaceae, Enterobacteriaceae and Comamonadaceae as the most abundant endophytes independently of land use intensity. Rhizosphere and bulk soil were dominated also by Proteobacteria, but the most abundant families differed from those obtained from root samples. In the soil, the effect of land use intensity was more pronounced compared to root endophytes leading to a clearly distinct pattern of bacterial communities under different LUI from rhizosphere and bulk soil vs. endophytes. Overall, a change of community structure on the plant–soil interface was observed, as the number of shared OTUs between all three compartments investigated increased with decreasing land use intensity. Thus, our findings suggest a stronger interaction of the plant with its surrounding soil under low land use intensity. Furthermore, the amount and quality of available nitrogen was identified as a major driver for shifts in the microbiome structure in all compartments

Relationships between soil physicochemical properties and nitrogen fixing, nitrifying and denitrifying under varying land-use practices in the northwest region of Argentina

The aim of this study was to evaluate the response pattern of diazotrophic microbes, denitrifiers and nitrifiers to different types of land use management, such as soybean monoculture (M) during 5 and 24 years (M5 and M24) and soybean-maize rotation (R) during 4 and 15 years (R4 and R15) in two subsequent years at the time point of flowering. Soil samples from a site recently introduced into agriculture (RUA) and a pristine soil under native vegetation (NV) were used as controls. Abundances of different functional groups of microbes were assessed using the direct quantification of marker genes by quantitative real-time PCR using extracted DNA from rhizosphere samples. In addition, soil chemical and physical properties were analysed and correlated with the abundance data from the functional microbial groups under investigation. Overall, the results indicate that the abundance of nifH genes was higher under R treatments compared to M treatments. The abundance of ammonium monooxygenase genes amoA (AOA) was generally higher under rotation systems and decreased under M24. RUA evidenced a negative effect on the establishment and development of AOA communities. The influence of land use on nirS abundance was inconsistent. However, R treatments showed a high abundance of nirK genes compared to M treatments. In both growing seasons, the abundance of nosZ genes was higher under NV compared with the other treatments. Furthermore, M24 treatment was related to strongly changed chemical and physical soil properties compared with the other sites. As expected, soil samples from RUA showed the strong dynamics of measured parameters indicating the high sensitivity of soils under transition to environmental parameters. Our results also indicated that the long-term crop rotation modified the abundance of the investigated microbial groups compared to the monoculture and increased soil chemical and physical quality. Therefore, our results provide evidence for a stimulatory effect of the long-term crop rotation on the abundance of microbes involved in N transformation.Fil: Perez Brandan, Carolina Gabriela. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Meyer, Annabel. Helmholtz Center Munich German Research Center For Environmental Health; AlemaniaFil: Meriles, Jose Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Huidobro, Jorgelina. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Schloter, Michael. Helmholtz Center Munich German Research Center For Environmental Health; AlemaniaFil: Vargas Gil, Silvina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Strukturelle und funktionelle Charakterisierung von mikrobiellen Gemeinschaften in ökologisch und konventionell bewirtschafteten Ackerböden

Strukturelle und funktionelle Charakterisierung von mikrobiellen Gemeinschaften in ökologisch und konventionell bewirtschafteten Ackerböden

Evaluation of denaturing gradient gel electrophoresis (DGGE) used to describe structure of bacterial communities in Istrian cheese

Denaturing gradient gel electrophoresis (DGGE) is a powerful method used to study structure of bacterial communities, without cultivation, based on the diversity of the genes coding for ribosomal RNA. However, the results are strongly dependent on the respective target region of the used primer systems. Therefore, three primer pairs that amplify different variable regions of the 16S rRNA gene (V1, V3 and V6 to V8) were tested in order to investigate the bacterial diversity existent in Istrian cheese. We found that primer set extremely influenced DGGE analysis. V3 primers were most efficient when 15 cheese associated isolates were resolved by DGGE. However, for Istrian cheese analysis, the best separation and highest number of bands in DGGE patterns were noticed for V6 to V8 primer pairs.Key words: Denaturing gradient gel electrophoresis, bacterial communities, Istrian cheese

Land use drives prokaryotic community composition of directly adjacent grasslands

Understanding the impact of agricultural land use on the soil prokaryotic communities in connected downslope sites is crucial for developing sustainable strategies to preserve ecosystem properties and mitigate agriculture’s environmental impacts. In this study, we investigated topsoil samples collected at three time points in 2022 (March, June, and November) from two adjacent catenas, reaching from hillslope to floodplain. The catenas differed in land use (extensive grassland vs. extensive cropland) at the top and middle parts, while the floodplain remained an extensive grassland due to legal restrictions. Using quantitative real-time PCRs and metabarcoding, we assessed prokaryotic abundance and prokaryotic community composition. Results show higher bacterial abundance in the cropland-influenced floodplain part across all time points compared to the grassland-influenced floodplain part. Temporal dynamics revealed a progressive decrease in the shared prokaryotic communities of the floodplain parts, peaking at the summer sampling time point, indicating a significant influence of the respective management type of the agricultural sites over the bacterial and archaeal communities of the floodplain parts. Differential abundance analyses identified several nitrifying taxa as more abundant in the cropland-influenced floodplain. Upstream land use also influenced the prokaryotic network of the cropland-floodplain, with some cropland taxa becoming keystone taxa and altering network morphology, an effect not observed in the grassland-influenced floodplain. These findings suggest that upstream agricultural land use practices have exerted a long-term influence on the floodplain prokaryotic communities over the past three decades. Moreover, there is evidence suggesting that these prokaryotic communities may undergo a potential reset during winter, which requires further investigation

Fire intensity regulates the short-term postfire response of the microbiome in Arctic tundra soil

Arctic tundra fires have been increasing in extent, frequency and intensity and are likely impacting both soil nitrogen (N) and phosphorus (P) cycling and, thus, permafrost ecosystem functioning. However, little is known on the underlying microbial mechanisms, and different fire intensities were neglected so far. To better understand immediate influences of different fire intensities on the soil microbiome involved in nutrient cycling in permafrost-affected soil, we deployed experimental fires with low and high intensity on an Arctic tundra soil on Disko Island, Greenland. Soil sampling took place three days postfire and included an unburned control. Using quantitative real-time PCR, copy numbers of 16S and ITS as well as of 17 genes coding for functional microbial groups catalyzing major steps of N and P turnover were assessed. We show that fires change the abundance of microbial groups already after three days with fire intensity as key mediating factor. Specifically, low-intensity fire significantly enhanced the abundance of chiA mineralizers and ammonia-oxidizing archaea, while other groups were not affected. On the contrary, high-intensity fire decreased the abundance of chiA mineralizers and of microbes that fix dinitrogen, indicating a dampening effect on N cycling. Only high-intensity fires enhanced ammonium concentrations (by an order of magnitude). This can be explained by burned plant material and the absence of plant uptake, together with impaired further N processing. Fire with high intensity also decreased nirK-type denitrifiers. In contrast, after fire with low intensity there was a trend for a decreased nosZ : (nirK+nirS) ratio, indicating – together with increased nitrate concentrations – an enhanced potential for nitric oxide and nitrous oxide emissions. Concerning P transformation, only gcd was affected in the short term which is important for P solubilization. Changes in gene numbers consistently showed the same contrasting pattern of elevated abundance with low fire intensity and decreased abundance with high fire intensity. Differentiating fire intensities is therefore crucial for further, longer-term studies of fire-induced changes in N and P transformations and potential nutrient-climate feedbacks of permafrost-affected soils

Site-Specific Conditions Change the Response of Bacterial Producers of Soil Structure-Stabilizing Agents Such as Exopolysaccarides and Lipopolysaccarides to Tillage Intensity

Agro-ecosystems experience huge losses of land every year due to soil erosion induced by poor agricultural practices such as intensive tillage. Erosion can be minimized by the presence of stable soil aggregates, the formation of which can be promoted by bacteria. Some of these microorganisms have the ability to produce exopolysaccharides and lipopolysaccharides that "glue" soil particles together. However, little is known about the influence of tillage intensity on the bacterial potential to produce these polysaccharides, even though more stable soil aggregates are usually observed under less intense tillage. As the effects of tillage intensity on soil aggregate stability may vary between sites, we hypothesized that the response of polysaccharide-producing bacteria to tillage intensity is also determined by site-specific conditions. To investigate this, we performed a high-throughput shotgun sequencing of DNA extracted from conventionally and reduced tilled soils from three tillage system field trials characterized by different soil parameters. While we confirmed that the impact of tillage intensity on soil aggregates is site-specific, we could connect improved aggregate stability with increased absolute abundance of genes involved in the production of exopolysaccharides and lipopolysaccharides. The potential to produce polysaccharides was generally promoted under reduced tillage due to the increased microbial biomass. We also found that the response of most potential producers of polysaccharides to tillage was site-specific, e.g., Oxalobacteraceae had higher potential to produce polysaccharides under reduced tillage at one site, and showed the opposite response at another site. However, the response of some potential producers of polysaccharides to tillage did not depend on site characteristics, but rather on their taxonomic affiliation, i.e., all members of Actinobacteria that responded to tillage intensity had higher potential for exopolysaccharide and lipopolysaccharide production specifically under reduced tillage. This could be especially crucial for aggregate stability, as polysaccharides produced by different taxa have different "gluing" efficiency. Overall, our data indicate that tillage intensity could affect aggregate stability by both influencing the absolute abundance of genes involved in the production of exopolysaccharides and lipopolysaccharides, as well as by inducing shifts in the community of potential polysaccharide producers. The effects of tillage intensity depend mostly on site-specific conditions

Endophytic root colonization of gramineous plants by Herbaspirillum frisingense

Herbaspirillum frisingense is a diazotrophic betaproteobacterium isolated from C4-energy plants, for example Miscanthus sinensis. To demonstrate endophytic colonization unequivocally, immunological labeling techniques using monospecific polyclonal antibodies against two H. frisingense strains and green fluorescent protein (GFP)-fluorescence tagging were applied. The polyclonal antibodies enabled specific in situ identification and very detailed localization of H. frisingense isolates Mb11 and GSF30T within roots of Miscanthus×giganteus seedlings. Three days after inoculation, cells were found inside root cortex cells and after 7 days they were colonizing the vascular tissue in the central cylinder. GFP-tagged H. frisingense strains could be detected and localized in uncut root material by confocal laser scanning microscopy and were found as endophytes in cortex cells, intercellular spaces and the central cylinder of barley roots. Concerning the production of potential plant effector molecules, H. frisingense strain GSF30T tested positive for the production of indole-3-acetic acid, while Mb11 was shown to produce N-acylhomoserine lactones, and both strains were able to utilize 1-aminocyclopropane-1-carboxylate (ACC), providing an indication of the activity of an ACC-deaminase. These results clearly present H. frisingense as a true plant endophyte and, although initial greenhouse experiments did not lead to clear plant growth stimulation, demonstrate the potential of this species for beneficial effects on the growth of crop plant