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At least 199 records · Page 11

Fate of Listeria monocytogenes Serotypes on Frozen Mixed Vegetables During Consumer‐Simulated Thawing and Storage

ABSTRACT Recent outbreaks and recalls associated with frozen vegetables in the United States and Europe have been linked to Listeria monocytogenes . This study aims to understand the extent to which frozen vegetables support the growth of L. monocytogenes once thawed and held at different temperatures. Six L. monocytogenes strains, two of each from serotypes 1/2a, 1/2b, and 4b, were individually inoculated onto frozen vegetables and stored at −18°C for 7 days. After 7 days, the vegetables were thawed and stored at 5°C or 10°C for up to 14 days or at 25°C for up to 7 days. L. monocytogenes was enumerated from the thawed vegetables throughout the storage period. Population data were fitted to the primary Baranyi model to estimate growth rates and lag phase durations; the secondary Ratkowsky square root model was used to model the relationship of the growth rates with storage temperature. Five of the L. monocytogenes strains survived and grew on the thawed vegetables (population increases of > 1 log CFU/g) stored at 5°C, and all six of the strains proliferated at 10°C and 25°C (population increases of > 3 log CFU/g after 14 days and > 4 log CFU/g after 7 days, respectively). A secondary model was successfully generated based on the growth rates of the six L. monocytogenes strains on the thawed vegetables ( r 2 = 0.8888, RMSE = 0.2057). Results from this study fill a data gap associated with L. monocytogenes survival on thawed vegetables and can be used to determine safe handling and storage practices for these products to protect public health.

Salazar, Joelle K. [Division of Food Processing Sc↗

Transcriptional, metabolic, physiological and developmental responses of switchgrass to phosphorus limitation

Abstract Knowing how switchgrass ( Panicum virgatum L . ) responds and adapts to phosphorus (P)‐limitation will aid efforts to optimize P acquisition and use in this species for sustainable biomass production. This integrative study investigated the impacts of mild, moderate, and severe P‐stress on genome transcription and whole‐plant metabolism, physiology and development in switchgrass. P‐limitation reduced overall plant growth, increased root/shoot ratio, increased root branching at moderate P‐stress, and decreased root diameter with increased density and length of root hairs at severe P‐stress. RNA‐seq analysis revealed thousands of genes that were differentially expressed under moderate and severe P‐stress in roots and/or shoots compared to P‐replete plants, with many stress‐induced genes involved in transcriptional and other forms of regulation, primary and secondary metabolism, transport, and other processes involved in P‐acquisition and homeostasis. Amongst the latter were multiple miRNA399 genes and putative targets of these. Metabolite profiling showed that levels of most sugars and sugar alcohols decreased with increasing P stress, while organic and amino acids increased under mild and moderate P‐stress in shoots and roots, although this trend reversed under severe P‐stress, especially in shoots.

59 BASIC BIOLOGICAL SCIENCES↗

University of Maryland Eastern Shore Common Garden

Located at the University of Maryland Eastern Shore, an HBCU (Historically Black Colleges and Universities), this data set is composed of the geographic coordinates of four transgenic Populus tremula x alba INRA 717-IB4 lines (CHX20, XBAT35, EXO70, DIR18) as well as a wild type (WT), and Empty vector (EV)). Our site is monitored by an ATOMS 41 weather station, which collects solar radiation, air temperature, vapor and atmospheric pressure, precipitation, and other weather variables. We will assess a suite of variables reflecting growth and performance, focusing on root allocation and soil carbon sequestration.

CBI, Populus, common garden, transgenic↗

Anti-fungal bioactive terpenoids in the bioenergy crop switchgrass (Panicum virgatum) may contribute to ecotype-specific microbiome composition

Abstract Plant derived bioactive small molecules have attracted attention of scientists across fundamental and applied scientific disciplines. We seek to understand the influence of these phytochemicals on rhizosphere and root-associated fungi. We hypothesize that – consistent with accumulating evidence that switchgrass genotype impacts microbiome assembly – differential terpenoid accumulation contributes to switchgrass ecotype-specific microbiome composition. An initial in vitro Petri plate-based disc diffusion screen of 18 switchgrass root derived fungal isolates revealed differential responses to upland- and lowland-isolated metabolites. To identify specific fungal growth-modulating metabolites, we tested fractions from root extracts on three ecologically important fungal isolates – Linnemania elongata, Trichoderma sp. and Fusarium sp. Saponins and diterpenoids were identified as the most prominent antifungal metabolites. Finally, analysis of liquid chromatography-purified terpenoids revealed fungal inhibition structure – activity relationships (SAR). Saponin antifungal activity was primarily determined by the number of sugar moieties – saponins glycosylated at a single core position were inhibitory whereas saponins glycosylated at two core positions were inactive. Saponin core hydroxylation and acetylation were also associated with reduced activity. Diterpenoid activity required the presence of an intact furan ring for strong fungal growth inhibition. These results inform future breeding and biotechnology strategies for crop protection with reduced pesticide application.

09 BIOMASS FUELS↗

Root responses to elevated CO 2 , warming and irrigation in a semi-arid grassland: Integrating biomass, length and life span in a 5-year field experiment

Plant roots mediate the impacts of environmental change on ecosystems, yet knowledge of root responses to environmental change is limited because few experiments evaluate multiple environmental factors and their interactions. Inferences about root functions are also limited because root length dynamics are rarely measured. Using a 5-year experiment in a mixed-grass prairie, we report the responses of root biomass, length and life span to elevated carbon dioxide (CO 2 ), warming, elevated CO 2 and warming combined, and irrigation. Root biomass was quantified using soil cores and root length dynamics were assessed using minirhizotrons. By comparing root dynamics with published results for soil resources and above-ground productivity, we provide mechanistic insights into how climate change might impact grassland ecosystems. In the upper soil layer, 0–15 cm depth, both irrigation and elevated CO 2 alone increased total root length by twofold, but irrigation decreased root biomass and elevated CO 2 had only small positive effects on root biomass. The large positive effects of irrigation and elevated CO 2 alone on total root length were due to increases in both root length production and root life span. The increased total root length and life span under irrigation and elevated CO 2 coincided with apparent shifts from water limitation of plant growth to nitrogen limitation. Warming alone had minimal effects on root biomass, length and life span in this shallow soil layer. Warming and elevated CO 2 combined increased root biomass and total root length by c. 25%, but total root length in this treatment was lower than expected if the effects of CO 2 and warming alone were additive. Treatment effects on total root length and root life span varied with soil depth and root diameter. Synthesis. Sub-additive effects of CO 2 and warming suggest studies of elevated CO 2 alone might overestimate the future capacity of grassland root systems to acquire resources. In this mixed-grass prairie, elevated CO 2 with warming stimulated total root length and root life span in deeper soils, likely enhancing plant access to more stable pools of growth-limiting resources, including water and phosphorus. Therefore, these root responses help explain previous observations of higher, and more stable, above-ground productivity in these projected climate conditions.

59 BASIC BIOLOGICAL SCIENCES↗

A single-cell Arabidopsis root atlas reveals developmental trajectories in wild-type and cell identity mutants

In all multicellular organisms, transcriptional networks orchestrate organ development. The Arabidopsis root, with its simple structure and indeterminate growth, is an ideal model to investigate the spatiotemporal transcriptional signatures underlying developmental trajectories. To map gene expression dynamics across root cell types and developmental time, we built a comprehensive, organ-scale atlas at single cell resolution. In addition to estimating developmental progressions in pseudotime, we employed the mathematical concept of optimal transport to infer developmental trajectories and identify their underlying regulators. To demonstrate the utility of the atlas to interpret new datasets, we profiled mutants for two key transcriptional regulators at single cell resolution, shortroot and scarecrow. We report transcriptomic and in vivo evidence for tissue trans-differentiation underlying a mixed cell identity phenotype in scarecrow. Our results support the atlas as a rich community resource for unraveling the transcriptional programs that specify and maintain cell identity to regulate spatiotemporal organ development.

59 BASIC BIOLOGICAL SCIENCES↗

Uptake and speciation of zinc in edible plants grown in smelter contaminated soils

Heavy metal accumulation in edible plants grown in contaminated soils poses a major environmental risk to humans and grazing animals. This study focused on the concentration and speciation of Zn in different edible plants grown in soils contaminated with smelter wastes (Spelter, WV, USA) containing high levels of the metals Zn, Cu, Pb, Cd. Their accumulation was examined in different parts (roots, stem, and leaves) of plants and as a function of growth stage (dry seed, sprouting seed, cotyledon, and leaves) in the root vegetables radish, the leafy vegetable spinach and the legume clover. Although the accumulation of metals varied significantly with plant species, the average metal concentrations were [Zn] > [Pb] > [Cu] > [Cd]. Metal uptake studies were complemented with bulk and micro X-ray absorption spectroscopy (XAS) at Zn K-edge and micro X-ray fluorescence (μXRF) measurements to evaluate the speciation and distribution of Zn in these plant species. Dynamic interplay between the histidine and malate complexation of Zn was observed in all plant species. XRF mapping of spinach leaves at micron spatial resolution demonstrated the accumulation of Zn in vacuoles and leaf tips. Radish root showed accumulation of Zn in root hairs, likely as ZnS nanoparticles. At locations of high Zn concentration in spinach leaves, μXANES suggests Zn complexation with histidine, as opposed to malate in the bulk leaf. These findings shed new light on the dynamic nature of Zn speciation in plants.

59 BASIC BIOLOGICAL SCIENCES↗

Methods and apparatus for gnotobiotic plant growth

The disclosure relates to a plant growth apparatus and related system to grow and maintain plants under controlled biotic conditions, for example to grow axenic (microbe-free) plants, gnotobiotic (defined microbiota) plants, and holoxenic (complex, or undefined microbiota) plants. This system allows aseptic bottom irrigation with water, soluble nutrients, chemicals, and/or microbiota. Plants can be inverted for dipping and/or vacuum infiltration. The system also allows for passive (gravity) drainage, thereby allowing for gas exchange and preventing root anoxia. A variety of plant growth substrates can be used within the plant growth apparatus as a plant growth medium. The plant growth apparatus, containing the growth substrate medium, can be completely flushed via the drainage port to remove potential toxic byproducts of the sterilization processes. The entire system is suitably constructed using autoclavable material.

59 BASIC BIOLOGICAL SCIENCES↗

Contrasting coordination of non‐structural carbohydrates with leaf and root economic strategies of alpine coniferous forests

Non-structural carbohydrates (NSCs), as the labile fraction and dominant carbon currency, are essential mediators of plant adaptation to environments. However, whether and how NSC coordinates with plant economic strategy frameworks, particularly the well-recognized leaf economics spectrums (LES) and root economics space (RES), remains unclear. We examined the relationships between NSC and key plant economics traits in leaves and fine roots across 90 alpine coniferous populations on the Tibetan Plateau, China. We observed contrasting coordination of NSC with economics traits in leaves and roots. Leaf total NSC and soluble sugar aligned with the leaf economic spectrum, conveying a trade-off between growth and storage in leaves. However, NSC in roots was independent of the root economic spectrum, but highly coordinated with root foraging, with more starch and less sugar in forage-efficient, thinner roots. Further, NSC-trait coordination in leaves and roots was, respectively, driven by local temperature and precipitation. These findings highlight distinct roles of NSC in shaping the above- and belowground multidimensional economics trait space, and NSC-based carbon economics provides a mechanistic understanding of how plants adapt to heterogeneous habitats and respond to environmental changes.

54 ENVIRONMENTAL SCIENCES↗

Phosphorus limitation of early growth differs between nitrogen‐fixing and nonfixing dry tropical forest tree species

Summary Tropical forests are often characterized by low soil phosphorus (P) availability, suggesting that P limits plant performance. However, how seedlings from different functional types respond to soil P availability is poorly known but important for understanding and modeling forest dynamics under changing environmental conditions. We grew four nitrogen (N)‐fixing Fabaceae and seven diverse non‐N‐fixing tropical dry forest tree species in a shade house under three P fertilization treatments and evaluated carbon (C) allocation responses, P demand, P‐use, investment in P acquisition traits, and correlations among P acquisition traits. Nitrogen fixers grew larger with increasing P addition in contrast to non‐N fixers, which showed fewer responses in C allocation and P use. Foliar P increased with P addition for both functional types, while P acquisition strategies did not vary among treatments but differed between functional types, with N fixers showing higher root phosphatase activity (RPA) than nonfixers. Growth responses suggest that N fixers are limited by P, but nonfixers may be limited by other resources. However, regardless of limitation, P acquisition traits such as mycorrhizal colonization and RPA were nonplastic across a steep P gradient. Differential limitation among plant functional types has implications for forest succession and earth system models.

59 BASIC BIOLOGICAL SCIENCES↗

Getting to the root of the problem: Soil carbon and microbial responses to root inputs within a buried paleosol along an eroding hillslope in southwestern Nebraska, USA

Large quantities of soil carbon (C) can persist within paleosols for millennia due to burial and subsequent isolation from plant-derived inputs, atmospheric conditions, and microbial activity at the modern surface. Erosion exposes buried soils to modern root-derived C influx via root exudation and root turnover, thus stimulating microbial activity leading to SOC decomposition and accumulation through organo-mineral stabilization of modern C. With this study we aim to quantify how modern root-derived C inputs impact paleosol C decomposition and stabilization across varying degrees of isolation from modern surface conditions in southwestern Nebraska, USA, where hillslope erosion is bringing a buried Late-Pleistocene-early Holocene paleosol (the “Brady Soil”) closer to the modern surface. We collected Brady Soil samples from 0.2m, 0.4m, and 1.2m below the modern surface and conducted two lab-based incubations. Soils were amended with either (1) a lab-synthesized mixture of low molecular weight compounds (12 atom% 13 C), or (2) 13 C enriched root residues (92 atom% 13 C), in 30-day and 240-day incubation experiments, respectively. Here we determined microbial responses to synthetic root exudates and residues by partitioning the 13 C label from Brady Soil C, including measurements of total, root, and primed C respiration, microbial biomass C (MBC), microbial C use efficiency (CUE). To assess the capacity of isolated paleosols to accrue modern plant C, we used Nano-scale Secondary Ion Mass Spectrometry imaging. We found that: (1) adding root-derived C inputs primed Brady Soil C across all depths, and was mediated by depth and composition of root additions; (2) root-derived C inputs stimulated microbial biomass C (MBC) growth similarly across depths, but the magnitude of CUE and MBC varied by chemistry of root-derived additions; (3) new particulate organic matter was incorporated into mineral-associated pools over time; (4) material from the added root residues was found in association with bacterial cells and fungal hyphae as well as with soil aggregate and mineral surfaces. Our study shows that paleosols defy expectations of C content and reactivity with depth, and changes in land cover and climate will expose buried paleosols to modern surface conditions, increasing respired C. This work highlights the importance of evaluating the role resurfacing buried soils through landscape change plays in C cycle feedbacks to the climate system.

54 ENVIRONMENTAL SCIENCES↗

Metabolome and RNA-seq Analysis of Responses to Nitrogen Deprivation and Resupply in Tea Plant (Camellia sinensis) Roots

Nitrogen (N) is an important contributor in regulating plant growth and development as well as secondary metabolites synthesis, so as to promote the formation of tea quality and flavor. Theanine, polyphenols, and caffeine are important secondary metabolites in tea plant. In this study, the responses of Camellia sinensis roots to N deprivation and resupply were investigated by metabolome and RNA-seq analysis. N deficiency induced content increase for most amino acids (AAs) and reduction for the remaining AAs, polyphenols, and caffeine. After N recovery, the decreased AAs and polyphenols showed a varying degree of recovery in content, but caffeine did not. Meanwhile, theanine increased in content, but its related synthetic genes were down-regulated, probably due to coordination of the whole N starvation regulatory network. Flavonoids-related pathways were relatively active following N stress according to KEGG enrichment analysis. Gene co-expression analysis revealed TCS2 , AMT1;1 , TAT2 , TS , and GOGAT as key genes, and TFs like MYB, bHLH, and NAC were also actively involved in N stress responses in C. sinensis roots. These findings facilitate the understanding of the molecular mechanism of N regulation in tea roots and provide genetic reference for improving N use efficiency in tea plant.

Xu, Wenluan↗

Uptake of polystyrene nanospheres by wheat and Arabidopsis roots in agar, hydroponics, and soil

Plant uptake of micro- and nanoplastics can lead to contamination of food with plastic particles and subsequent human consumption of plastics. There is evidence that plant roots can take up micro and nanoplastics; however, most of this evidence stems from experiments conducted with plants grown in hydroponics or agar systems where uptake of nanoparticles by roots is more favorable than when plants were grown in soil. Here, we discern the root uptake and accumulation of polystyrene nanospheres in plants grown in different growth media: agar, hydroponics, and soil. In addition, we tested the impacts of nanospheres on plant biomass and plant stress. Wheat and Arabidopsis thaliana were grown in agar, hydroponics, and soil media and exposed to polystyrene nanospheres. Three different nanospheres were used (40 nm and 200 nm carboxylate-modified and 200 nm amino-modified polystyrene) and uniformly mixed into the growth media. Plants were grown for 7 to 10 days and the roots were then examined for the presence of nanospheres by confocal laser scanning microscopy and scanning electron microscopy. Plant stress was evaluated by measuring reactive oxygen species (ROS). We observed the 40 nm nanospheres inside the plant roots, but the 200 nm nanospheres only adhered to the root cap cells showing no uptake into the roots. Furthermore, confocal images indicated that root uptake of nanospheres was favored in hydroponic solutions as compared to agar and soil media. Plant biomass was generally not affected by the nanospheres, except for hydroponically grown Arabidopsis thaliana, where biomass was significantly reduced. Small sized (40 nm) and positively charged (200 nm amino-modified) nanospheres showed higher ROS accumulation in plants than negatively charged 200 nm carboxylate-modified nanospheres. In conclusion, this study provides evidence that polystyrene nanospheres can be taken up into the interior of plant roots and cause plant stress, but these impacts are less pronounced in media where the plastic particles are less mobile, like in agar and soil media as compared to hydroponic systems.

54 ENVIRONMENTAL SCIENCES↗

Size-related decline in dryland shrubs is related to reductions in hydraulic efficiency and carbon assimilation and not nonstructural carbohydrate depletion

Plant growth and survival are fundamentally constrained by water transport from roots to leaves, impacting carbon assimilation and associated labile carbon pools. However, physiological constraints on growth and survival vary with plant age, due to changes in metabolic sinks, and increases in hydraulic path length from rhizosphere to canopy. We investigated crown dieback, growth, hydraulics, carbon assimilation and carbohydrate storage in relation to increasing basal diameter of two dominant shrub species (Caragana korshinskii and Artemisia ordosica) at the southeastern edge of the Tengger Desert, China. The aim was to identify mechanisms of decreased performance with plant size in dryland shrubs. Clear contrasts in stomatal regulation of leaf water potentials were detected between species. Despite these contrasts, radial growth, hydraulic transport efficiency (Ks), and photosynthetic efficiency similarly declined in both species with increasing plant size, while non-structural carbohydrate (NSC) reserves remained unchanged. Xylem embolism (PLC) increased with plant size, resulting in significant reductions in carbon assimilation in both species. Results indicate that hydraulic, and potentially carbon assimilation constraints, rather than reductions in carbohydrate storage, govern growth-related dryland shrub decline. These findings improve our understanding of how population demography impacts dryland forest response to climate change.

Zhang, Hongxia↗

Review of Landscaping Equipment Vibration Risk Exposure, LLNL Site 200 (Various Locations), Whole Body Vibration Seated and Standing Mowers

WBV (Whole-Body Vibration) for operators of seated and standing lawn mowers was conducted during landscape activities from April 2019 through August 2019 at various locations around LLNL Site 200. Mowing is generally performed by each landscaper for 2-3 days every other week with a maximum duration of 3-5 hours each day. On mowing days, other tasks are performed for the remainder of the day that do not expose the worker to WBV such as irrigation, blowing, or sweeping. The mowing terrain at LLNL is variable with typical reported conditions such as gopher holes, exposed tree roots, quick turns, normal and high growth, weeds, and mushy/wet ground that can result in jolts to the mower operator. The Larson Davis HVM200 seat pad with tri-axial accelerometer was used for monitoring workers. For the seated mowers, the HVM200 seat pad with tri-axial accelerometer was placed on the seat between the user and seat pan of the mower. For standing mowers, the seat pad with tri-axial accelerometer was wrapped with Dycem (for seat pad protection) and placed directly on top of the mower deck and underneath the end-user’s right foot. Monitoring for WBV was conducted while operators used different types, makes, and models of mowers. A total of sixteen samples were collected from the operators of seated mower and 6 samples were collected from operators of standing mower. This initial data sampling was part of a discovery phase to identify if WBV exposure met the 2016 ACGIH TLVs (Threshold Limit Values). This initial data identified that workers operating the mowers do not exceed either the TLV or Action Level (AL) for Weighted Acceleration Levels (A(8)) but typically exceed the Vibration Dose Value (VDV) lower boundary for the standing mowers.

59 BASIC BIOLOGICAL SCIENCES↗

Biomolecular and Characterization Imaging Science Program: 2023 Principal Investigator Meeting Proceedings

The 2023 U.S. Department of Energy (DOE) Biological and Environmental Research (BER) program’s Biomolecular Characterization and Imaging Science (BCIS) Principal Investigator Meeting expanded in scope from previous Bioimaging Science Program (BSP) meetings to include BER Structural Biology and Imaging Resources, which are located largely at DOE Office of Science national laboratories. The BCIS meeting was part of BER’s Biological Systems Science Division (BSSD) annual PI meeting, which was held April 17–19, 2023, and featured parallel meetings of the BCIS and Genomic Science programs (GSP). The meetings were held together to encourage networking and idea exchange across technologies and biological application areas, forging new multidisciplinary collaborations among researchers from adjacent BSSD programmatic areas. Two joint BCIS-GSP sessions were held: “BCIS Technologies for Investigating the Rhizosphere” and “Joint Emerging Topics and Technologies.” The rhizosphere session focused on scientific findings from BCIS and GSP PIs, including national laboratory collaborations. The intent was to identify new opportunities to measure and understand the complex community of microbes, roots, and soils that support plant growth under challenging environmental conditions. The emerging technologies session highlighted forward-looking approaches and tools to tackle challenges within the scope of BSSD research on investigating and modifying genomic and molecular function. A final interactive discussion of the BCIS program was led by plenary session chairs.

59 BASIC BIOLOGICAL SCIENCES↗

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↗