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At least 415 records · Page 23

Anaerobic Psychrophiles from Lake Zub and Lake Untersee, Antarctica

The study of samples from Antarctica 2008 and 2009 expeditions organized and successfully conducted by Richard Hoover led to the isolation of diverse anaerobic strains with psychrotolerant and psychrophilic physiology. Due to the fact that Lake Untersee has never been subject to microbiological study, this work with the samples has significant and pioneering impact to the knowledge about the biology of this unique ecosystem. Also, the astrobiological significance for the study of these ecosystems is based on new findings of ice covered water systems on other bodies of our solar system. Anaerobic psychrotolerant strain LZ-22 was isolated from a frozen sample of green moss with soils around the rhizosphere collected near Lake Zub in Antarctica. Morphology of strain LZ-22 was observed to be motile, rod shaped and spore-forming cells with sizes 1 x 5-10 micron. This new isolate is a mesophile with the maximum temperature of growth at 40C. Strain LZ-22 is able to live on media without NaCl and in media with up to 7% (w/v) NaCl. It is catalase negative and grows only on sugars with the best growth rate being on lactose. The strain is a neutrophile and grows between pH 5 and 9.0 with the optimum at 7.8. Another two strains UL7-96mG and LU-96m7P were isolated from deep water samples of Lake Untersee. Proteolytic strain LU-96m7P had a truly psychrophilic nature and refused to grow at room temperature. Sugarlytic strain UL7-96mG was found to be psychrotolerant, but its rate of growth at 3C was very high compared with other mesophiles. Two homoacetogenic psychrophilic strains A7AC-96m and AC-DS7 were isolated and purified from samples of Lake Untersee; both of them are able to grow chemolithotrophically on H2+CO2. In the presence of lactate, these strains are able to grow only at 0-18C, and growth at 22C was observed only with yeast extract stimulation. In this paper, physiological and morphological characteristics of novel psychrophilic and psychrotolerant isolates from Antarctica 2008 and 2009 expeditions will be discussed.

Townsend, Alisa↗

Plant Microbiomes May Provide Vital Information to Plant Success

Plant associated microbiomes, the rhizosphere and phyllosphere, are composed of communities of bacteria and fungi that may be mutualistic or pathogenic. These communities have the potential to influence plant health and development and can affect plant growth. Crop plants are being investigated as a fresh and safe supplement to astronauts’ diet and it is critical to understand and characterize these microbial communities. Multi-species crops, Mizuna mustard (Brassica rapa var japonica), ‘Outredgeous’ red romaine lettuce (Lactuca sativa), and Waldman’s Green lettuce (Lactuca sativa) were grown in two Veggie units on the International Space Station (ISS) for three grow outs in various combinations of plant types. Upon harvest, plant and pillow samples were frozen and returned to Earth for analysis. Bacterial and fungal community analyses for plant leaf and root, as well as pillow components, wick and media, were completed using next generation sequencing with the goal of surveying the composition of the entire community and identifying any potential pathogens. Bacteria were identified using the 16S rRNA gene whereas, fungi were identified with the internal transcribed spacer (ITS). The community composition for these three crops was compared between crop types and between plant tissue types. It is vital to mission success for the short term and long term to add nutritious, safe to eat vegetables providing a supplement to the crew members’ dietary requirements as well as to develop planning for deep space missions as we reach for the moon and on to Mars. Veggie technology validation tests were supported by NASA’s Space Biology Program.

Khodadad, Christina L.↗

Microbial sensor system for monitoring and imaging of an environment

A microbial sensor, microbial sensing system, and method that can be used to determine the chemical environment of unsaturated soils, rhizosphere, and/or plants are disclosed. The microbial sensing system can be used for monitoring the health of plants including nutrients, salinity, contaminants, chemicals (pesticides, herbicides) and diseases. A microbial sensing system can include one or more indicator electrodes and a reference electrode. The microbial sensing system can include a signal acquisition and/or communication module to allow the real-time collection of data from field deployments and laboratory investigations.

Burge, Scott R.↗

Microbial sensor system for monitoring the environment and surfaces

A microbial sensor, system and method that can be used to determine the biochemical conditions of waters, saturated sediments, unsaturated soils, rhizosphere and other environments is disclosed. The flexibility of the microbial sensor design allows for the monitoring of surfaces (skin, ventilation conduits, etc.). An electrochemical microbial sensor system is composed of an indicator electrode(s) and a reference electrode. The reference electrode employs a hydrogen permeable membrane. The electrochemical system is interfaced into a signal/communication module allowing the manual or automated collection of data from field deployments and laboratory investigations. The data is transmitted using various communication technologies including Bluetooth™, cellular, satellite and radio telemetry to cloud-based data management systems. The stored data may be downloaded by users using open-source dashboard and visualization software to image the various environments and/or surfaces investigated.

Burge, Scott R.↗

Capillary absorption spectroscopy for high temporal resolution measurements of stable carbon isotopes in soil and plant-based systems

Capillary Absorption Spectroscopy (CAS) is a relatively new analytical technique for performing stable isotope analysis. Here, we demonstrate the utility of CAS by recording and quantifying variation in 13C in controlled and biologically relevant applications. We calibrated CAS system response to increased 13CO2, with an observed ~4‰ increase in measured Δ13C for each 0.03 ppm shift in 13CO2 concentration. We leveraged this calibration to quantify rates of biogeochemical processes using a 13C tracer. For example, we monitored microbial respiration of 13C-glucose within an agricultural soil at 10 s quantification intervals and results demonstrated 8.6% ± 0.4 of added glucose was converted to 13CO2 within 1.5 h of incubation. We expanded the demonstration by adapting a rhizobox to permit continuous monitoring of 13CO2 in a soil (as distinct from plant) headspace to track the timing and quantify respiration rates of fresh plant photosynthate and observed a 3.5 h delay between plant exposure to a13CO2 tracer and the first signs of respiration by soil biota. These experiments highlight CAS is effective in producing high temporal resolution quantification of 13CO2 and demonstrate potential applications.

13CO2↗

The nitrogen gap in soil health concepts and fertility measurements

Soil nitrogen (N) often limits productivity in agroecosystems, prompting fertilizer applications that increase crop yields but can degrade the environment. Nitrogen's dual role in both productivity and environmental quality should center it in soil health frameworks. We use recent evidence to argue that N availability is an emergent property of the integrated soil biogeochemical system and is strongly influenced by plant traits and their interactions with microbes and minerals. Building upon this, we theorize that the sources of plant and microbial N shift across soil health gradients, from inorganic N dependence in ecologically simple systems with poor soil health to a highly networked supply of organic N in healthy soils; ergo, investments in soil health should increase ecological complexity and the pathways by which plants can access N, leading to more resilient nutrient supplies and yields in a variable climate. However, current N assessment methods derive from a historical emphasis on inorganic N pool sizes and are unable to capture the shifting drivers of N availability across soil health gradients. We highlight the need to better understand the plant-microbial-mineral interactions that regulate bioavailable N as a first step to improving our ability to measure it. We conclude it will be necessary to harness agroecosystem complexity, account for plant and microbial drivers, and carefully integrate external N inputs into soils' internal N network to expand the routes by which N from organic pools can be made bioavailable. Finally, by emphasizing N in soil health concepts, we argue that researchers can accelerate advances in N use efficiency and resiliency.

60 APPLIED LIFE SCIENCES↗

Coastal Forest Seawater Exposure Increases Stem Methane Concentration

Methane (CH 4 ) exchange between trees and the atmosphere has recently emerged as an important, but poorly quantified process regulating global climate. The sources (soil and/or tree) and mechanisms driving the increase of CH 4 in trees and degassing to the atmosphere are inadequately understood, particularly for coastal forests facing increased exposure to seawater. Here, we investigated the eco-physiological relationship between tree stem wood density, soil and stem oxygen saturation (an indicator of redox state), soil and stem CH 4 concentrations, soil and stem carbon dioxide (CO 2 ) concentrations, and soil salinity in five forests along the United States coastline. We aim to evaluate the mechanisms underlying greenhouse gas (GHG) increase in trees and the influence of seawater exposure on stem CH 4 accumulation. Seawater exposure corresponded with decreased tree survival and increased tree stem methane. Tree stem wood density was significantly correlated with increased stem CH4 in seawater exposed gymnosperms, indicating that dying gymnosperm trees may accumulate higher levels of CH 4 in association with seawater flooding. Further, we found that significant differences in seawater exposed and unexposed gymnosperm tree populations are associated with increased soil and stem CH 4 and CO 2 , indicating that seawater exposure significantly impacts soil and stem greenhouse gas abundance. Our results provide new insight into the potential mechanisms driving tree CH 4 accumulation within gymnosperm coastal forests.

54 ENVIRONMENTAL SCIENCES↗

Accounting for linkages between wildfire-driven shifts in plant-microbial interactions and soil carbon dynamics in Arctic tundra

Increasing wildfire regimes in the rapidly changing Arctic tundra are altering the soil carbon budget through increased permafrost degradation, shrubs expansion, and shifts in microbial activities. Whether future arctic wildfires will result in net C losses or gains in the future will depend on complex biotic and abiotic interactions that regulate belowground C biogeochemical processes, including linkages among biotic communities. One important linkage is plant-microbe interactions. While these interactions are likely shaped or altered by wildfires, they remain little explored in the context of successional trajectories. Yet, incorporating plant-microbe interactions in frameworks for defining and understanding post-fire soil C trajectories is critical to predict belowground C responses to future tundra wildfires. Here, we provide examples of and discuss how fire-mediated changes in plant-soil-microbe (PSM) interactions can impact soil C dynamics in the Arctic tundra. We consider different impacts of wildfires on PSM interactions and their implications to soil C dynamics, as well as the nuances associated with particular wildfire regimes (severity and intensity) and successional timescales. We suggest that accounting for plant-microbial linkages in future wildfire-succession interactions frameworks can inform future experimental designs and reduce uncertainties in our ability to predict the net effect of Arctic wildfires on ecosystem C.

fungi, bacteria↗

RhizoVision Explorer: open-source software for root image analysis and measurement standardization

Abstract Roots are central to the function of natural and agricultural ecosystems by driving plant acquisition of soil resources and influencing the carbon cycle. Root characteristics like length, diameter and volume are critical to measure to understand plant and soil functions. RhizoVision Explorer is an open-source software designed to enable researchers interested in roots by providing an easy-to-use interface, fast image processing and reliable measurements. The default broken roots mode is intended for roots sampled from pots and soil cores, washed and typically scanned on a flatbed scanner, and provides measurements like length, diameter and volume. The optional whole root mode for complete root systems or root crowns provides additional measurements such as angles, root depth and convex hull. Both modes support providing measurements grouped by defined diameter ranges, the inclusion of multiple regions of interest and batch analysis. RhizoVision Explorer was successfully validated against ground truth data using a new copper wire image set. In comparison, the current reference software, the commercial WinRhizo™, drastically underestimated volume when wires of different diameters were in the same image. Additionally, measurements were compared with WinRhizo™ and IJ_Rhizo using a simulated root image set, showing general agreement in software measurements, except for root volume. Finally, scanned root image sets acquired in different labs for the crop, herbaceous and tree species were used to compare results from RhizoVision Explorer with WinRhizo™. The two software showed general agreement, except that WinRhizo™ substantially underestimated root volume relative to RhizoVision Explorer. In the current context of rapidly growing interest in root science, RhizoVision Explorer intends to become a reference software, improve the overall accuracy and replicability of root trait measurements and provide a foundation for collaborative improvement and reliable access to all.

59 BASIC BIOLOGICAL SCIENCES↗

Microbial community dynamics in the soil-root continuum are linked with plant species turnover during secondary succession

Grazing exclusion and land abandonment are commonly adopted to restore degraded ecosystems in semiarid and arid regions worldwide. However, the temporal variation in the soil- versus root-associated microbiome over plant species turnover during secondary succession has rarely been quantified. Using the chronosequence restored from fenced grassland and abandoned farmlands on the Loess Plateau of China, we characterized the dynamics of the soil- and root-associated microbiome of host plant with different dominance statuses during secondary succession from 0 to 40 years. Our results revealed that the root microhabitat, the host plant and their interactions were the main contributors to the bacterial community shift (R 2 = 15.5%, 8.1%, and 22.3%, respectively), and plant interspecies replacement had a greater effect on the shift in the root-associated microbial community than intraspecies replacement did during succession. The root-associated bacterial community of pioneer plants was particularly responsive to succession, especially the endosphere community. Endosphere microbial diversity was positively correlated with host plant coverage change, and the diversity and abundance of taxon recruitment into the endosphere of pioneer plants from the surrounding environment decreased as succession progressed. The community assembly processes also indicated that the endosphere microbiota are strongly selected in younger host plants, whereas stochastic processes dominate in aged host plants. Our study provides evidence of the unique response of the root-associated microbiome to the replacement of plant species during secondary succession, and the function of endosphere microbes should be considered when studying plant–microbe feedback.

59 BASIC BIOLOGICAL SCIENCES↗

Phenylpropanoid methyl esterase unlocks catabolism of aromatic biological nitrification inhibitors

Microbial nitrification of fertilizers represents is a significant global source of greenhouse gas emissions. This process increases emissions, fosters toxic algal blooms, and raises crop production costs. Some plants naturally release biological nitrification inhibitors to suppress ammonium-oxidizing microbes and reduce nitrification. Engineering nitrification inhibitor production into food and bioenergy crops via synthetic biology offers a promising mitigation strategy, but its success depends on addressing gaps in our understanding of inhibitor degradation in soil. This study begins to fill this gap by identifying a previously unknown microbial pathway for degrading phenylpropanoid methyl esters, a key class of aromatic nitrification inhibitors. Using transcriptomics and high-throughput functional genomics, we discovered genes essential for phenylpropanoid methyl ester degradation. Genetic and biochemical analyses revealed two novel enzymes, including a newly identified phenylpropanoid methyl esterase, that direct phenylpropanoid methyl esters into known metabolic pathways. Importantly, transferring these genes into bacteria capable of metabolizing other phenylpropanoids enabled them to use the methyl esters as a carbon source. This work provides critical insights into microbial nitrification inhibitor degradation, a poorly understood element of the nitrification cycle.

Genetic Engineering↗

Development of modular expression across phylogenetically distinct diazotrophs

Diazotrophic bacteria can reduce atmospheric nitrogen into ammonia enabling bioavailability of the essential element. Many diazotrophs closely associate with plant roots increasing nitrogen availability, acting as plant growth promoters. These associations have the potential to reduce the need for costly synthetic fertilizers if they could be engineered for agricultural applications. However, despite the importance of diazotrophic bacteria, genetic tools are poorly developed in a limited number of species, in turn narrowing the crops and root microbiomes that can be targeted. Here, we report optimized protocols and plasmids to manipulate phylogenetically diverse diazotrophs with the goal of enabling synthetic biology and genetic engineering. Three broad-host-range plasmids can be used across multiple diazotrophs, with the identification of one specific plasmid (containing origin of replication RK2 and a kanamycin resistance marker) showing the highest degree of compatibility across bacteria tested. We then demonstrated modular expression by testing seven promoters and eleven ribosomal binding sites using proxy fluorescent proteins. Finally, we tested four small molecule inducible systems to report expression in three diazotrophs and demonstrated genome editing in Klebsiella michiganensis M5al.

59 BASIC BIOLOGICAL SCIENCES↗

The Switchgrass Microbiome: A Review of Structure, Function, and Taxonomic Distribution

Switchgrass (Panicum virgatum L.) has been championed as a promising bioenergy crop due to its high productivity across a wide environmental range. The switchgrass microbiome—including bacteria, archaea, fungi, and other microbiota inhabiting soil and plant tissues—can influence plant function substantially. We conducted a review of the literature investigating switchgrass microbiome structure, key functional roles, and taxa isolated from field-grown plants. Although site conditions and plant compartment (i.e., location within shoots, roots, or root-influenced soil) appear to be the strongest drivers of switchgrass microbiome structure, the microbiome is also shaped by climate, season, and host genotype. Studies comparing across plant species show that the switchgrass microbiome is more similar to the microbiomes of other perennial plants than to the microbiomes of annual plants. Members of the switchgrass microbiome confer several benefits to their host. Most notably, mycorrhizal fungi can increase plant biomass many-fold, associative nitrogen-fixing bacteria can provide a substantial portion of the plant’s nitrogen demand, and fungal endophytes can improve plant tolerance to drought. Although the fungi and bacteria cultured from switchgrass represent only a portion of the microbiome, these serve as a valuable resource for researchers interested in investigating functional outcomes of the switchgrass microbiome. We highlight areas where additional research is necessary for a more comprehensive understanding of switchgrass microbiome structure, function, and potential to enhance sustainable bioenergy production. Key gaps include the role of understudied organisms (e.g., viruses, microeukaryotes, and nonmycorrhizal fungi), multitrophic relationships, mechanisms underpinning switchgrass–microbiome interactions, and field-scale validation of experimental findings.

54 ENVIRONMENTAL SCIENCES↗

Novel and Emerging Capabilities that Can Provide a Holistic Understanding of the Plant Root Microbiome

In recent years, the root microbiome (i.e., microorganisms growing inside, on, or in close proximity to plant roots) has been shown to play an important role in plant health and productivity. Despite its importance, the root microbiome is challenging to study because of its complexity, heterogeneity, and subterranean location. Fortunately, root microbiome research has seen a tremendous influx of novel technologies (e.g., imaging tools, robotics, and molecular analyses), experimental platforms (e.g., micro- and mesocosms), and data integration, modeling, and prediction tools in the past decade that have greatly increased our ability to dissect the complex network of interactions between above- and belowground environmental parameters, plants, bacteria, and fungi that dictate soil and broader ecosystem health. Herein, we discuss methods that are currently used in root microbiome research and that can be expanded to phytobiome research in general ranging from laboratory studies to mesocosm-scale studies and, finally, to field studies; evaluate their relevance to ecosystem studies; and discuss future root microbiome research directions.

59 BASIC BIOLOGICAL SCIENCES↗