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DOE OSTI2021

A critical review of molybdenum sequestration mechanisms under euxinic conditions: Implications for the precision of molybdenum paleoredox proxies

The observed difference in molybdenum (Mo) mobility and isotopic fractionation under oxic versus euxinic (i.e., anoxic and sulfidic) aqueous conditions provides a sound operational basis for the use of Mo geochemical signatures in ancient sedimentary records to infer palaeoceanographic redox conditions. While Mo is known to exist predominantly as molybdate (MoO 4 2– ) in oxic waters and convert into thiomolybdate species (MoO x S 4-x 2– ) under euxinic conditions, the pathways that lead to Mo sequestration are highly debated. As mechanistic understanding of Mo sequestration is crucial for accurately reconstructing the chemistry of ancient oceans and constraining the timing and intensity of oxygenation events through Earth's history using Mo paleoproxies, we have closely examined the current proposed mechanisms for Mo sequestration across a wide range of euxinic conditions. Through compilation and comparison of such information, we aim to provide an integrated view of Mo sequestration processes, to identify the current controversies as well as the roots of such controversies, and importantly, to propose avenues for future research. Here, sequestration of Mo may occur through complexation with organic matter (OM), reactions with oxyhydroxide and sulfide mineral surfaces, incorporation into iron sulfide crystal structures, formation of Mo-sulfide, and cellular assimilation. However, several major questions remains unresolved, including whether OM complexation plays a significant role in overall sedimentary Mo accumulation, whether the reduction of Mo(VI) occurs during interaction with Fe-S clusters/precipitates and/or with OM, whether microbial enzymatic reduction of Mo(VI) is important in Mo sequestration, whether synergistic effects exist between the biological and abiotic processes in Mo sequestration, and whether differences in reactivity among the various thiomolybdate intermediate species and tetrathiomolybdate influence the Mo sequestration process. At the end of the review, the reliability of Mo paleoproxies is discussed and reevaluated.

58 GEOSCIENCES↗

DOE OSTI2019

High spatial resolution imaging of biological tissues using nanospray desorption electrospray ionization mass spectrometry

Mass spectrometry imaging (MSI) enables label-free spatial mapping of hundreds of biomolecules in tissue sections. This capability provides valuable information on tissue heterogeneity that is difficult to obtain using population-averaged assays. Despite substantial developments in both instrumentation and methodology, MSI of tissue samples at single-cell resolution remains challenging. In our research, we describe a protocol for robust imaging of tissue sections with a high (better than 10-μm) spatial resolution using nanospray desorption electrospray ionization (nano-DESI) mass spectrometry, an ambient ionization technique that does not require sample pretreatment before analysis. In this protocol, mouse uterine tissue is used as a model system to illustrate both the workflow and data obtained in these experiments. We provide a detailed description of the nano-DESI MSI platform, fabrication of the nano-DESI and shear force probes, shear force microscopy experiments, spectral acquisition, and data processing. A properly trained researcher (e.g., technician, graduate student, or postdoc) can complete all the steps from probe fabrication to data acquisition and processing within a single day. We further describe a new strategy for acquiring both positive- and negative-mode imaging data in the same experiment. This is achieved by alternating between positive and negative data acquisition modes during consecutive line scans. Using our imaging approach, hundreds of high-quality ion images were obtained from a single uterine section. This protocol enables sensitive and quantitative imaging of lipids and metabolites in heterogeneous tissue sections with high spatial resolution, which is critical to understanding biochemical processes occurring in biological tissues.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2019

What do we know about soil carbon destabilization?

Abstract Most empirical and modeling research on soil carbon (C) dynamics has focused on those processes that control and promote C stabilization. However, we lack a strong, generalizable understanding of the mechanisms through which soil organic carbon (SOC) is destabilized in soils. Yet a clear understanding of C destabilization processes in soil is needed to quantify the feedbacks of the soil C cycle to the Earth system. Destabilization includes processes that occur along a spectrum through which SOC shifts from a ‘protected’ state to an ‘available’ state to microbial cells where it can be mineralized to gaseous forms or to soluble forms that are then lost from the soil system. These processes fall into three general categories: (1) release from physical occlusion through processes such as tillage, bioturbation, or freeze-thaw and wetting-drying cycles; (2) C desorption from soil solids and colloids; and (3) increased C metabolism. Many processes that stabilize soil C can also destabilize C, and C gain or loss depends on the balance between competing reactions. For example, earthworms may both destabilize C through aggregate destruction, but may also create new aggregates and redistribute C into mineral horizon. Similarly, mycorrhizae and roots form new soil C but may also destabilize old soil C through priming and promoting microbial mining; labile C inputs cause C stabilization through increased carbon use efficiency or may fuel priming. Changes to the soil environment that affect the solubility of minerals or change the relative surfaces charges of minerals can destabilize SOC, including increased pH or in the reductive dissolution of Fe-bearing minerals. By considering these different physical, chemical, and biological controls as processes that contribute to soil C destabilization, we can develop thoughtful new hypotheses about the persistence and vulnerability of C in soils and make more accurate and robust predictions of soil C cycling in a changing environment.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2019

Integrated Omic Analyses Provide Evidence that a “ Candidatus Accumulibacter phosphatis” Strain Performs Denitrification under Microaerobic Conditions

“ Candidatus Accumulibacter phosphatis” is widely found in full-scale wastewater treatment plants, where it has been identified as the key organism for biological removal of phosphorus. Since aeration can account for 50% of the energy use during wastewater treatment, microaerobic conditions for wastewater treatment have emerged as a cost-effective alternative to conventional biological nutrient removal processes. Our report provides strong genomics-based evidence not only that “ Ca . Accumulibacter phosphatis” is the main organism contributing to phosphorus removal under microaerobic conditions but also that this organism simultaneously respires nitrate and oxygen in this environment, consequently removing nitrogen and phosphorus from the wastewater. Such activity could be harnessed in innovative designs for cost-effective and energy-efficient optimization of wastewater treatment systems.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2025

Algae Biofilm Produced from Anaerobic Digester Wastewater Demonstrated as Biofertilizer for Dwarf Wheat

Aims: To correlate yield of dwarf wheat fertilized with algae biofilm produced using anaerobic digester effluent (digestate) from a municipal water resource recovery facility as nutrient source and compare with yield using non-algae biofilm based fertilizers and a negative control. Study Design: Greenhouse testing for measurement of biomass yields and statistical analysis of wheat yield data results. Place and Duration of Study: Department of Biological Engineering Algae Processing and Products Facility greenhouse between March and June, 2024. Methodology: Yield of a dwarf cultivar of red spring wheat was measured as a function of four treatments including three fertilizers and a negative control: (1) algae biofilm cultivated on anaerobic digester effluent, (2) positive control of Osmocote commercial slow release fertilizer, (3) anaerobic digester biosolids, and (4) negative control with no fertilizer addition. Results: Average yield values (gm) for the four treatments were: (1) 1.25, (2) 0.99, (3) 0.72, and (4) 0.52, respectively. Algae biofilm fertilizer performed significantly better than Osmocote with P = .002, anaerobic digester biosolids with P = .05, and no fertilizer addition with P = .05. Struvite, a slow release fertilizer, was observed to be associated with the algae biofilm as a precipitate of the high concentrations of nutrients within the anaerobic digester water. Struvite (MgNH4PO46H2O) releases plant available nitrogen and phosphorus over time and is biologically precipitated due to the increase in solution pH as a result of photosynthesis by the algae biofilm. Conclusion: Algae biofilm cultivated on anaerobic digester wastewater can serve as a biofertilizer for recycling nutrients within the biosphere for sustainable nutrient management.

09 BIOMASS FUELS↗

DOE OSTI2019

Anisohydric behavior linked to persistent hydraulic damage and delayed drought recovery across seven North American tree species

Summary The isohydry‐anisohydry spectrum has become a popular way to characterize plant drought responses and recovery processes. Despite the proven utility of this framework for understanding the interconnected physiological changes plants undergo in response to water stress, new challenges have arisen pertaining to the traits and tradeoffs that underlie this concept. To test the utility of this framework for understanding hydraulic traits, drought physiology and recovery, we applied a 6 wk experimental soil moisture reduction to seven tree species followed by a 6 wk recovery period. Throughout, we measured hydraulic traits and monitored changes in gas exchange, leaf water potential, and hydraulic conductivity. Species’ hydraulic traits were not coordinated, as some anisohydric species had surprisingly low resistance to embolism (P 50 ) and negative safety margins. In addition to widespread hydraulic damage, these species also experienced reductions in photosynthesis and stem water potential during water stress, and delayed recovery time. Given that we observed no benefit of being anisohydric either during or after drought, our results indicate the need to reconsider the traits and tradeoffs that underlie anisohydric behavior, and to consider the environmental, biological and edaphic processes that could allow this strategy to flourish in forests.

Kannenberg, Steven A.↗

NASA NTRS1994

From neural-based object recognition toward microelectronic eyes

Engineering neural network systems are best known for their abilities to adapt to the changing characteristics of the surrounding environment by adjusting system parameter values during the learning process. Rapid advances in analog current-mode design techniques have made possible the implementation of major neural network functions in custom VLSI chips. An electrically programmable analog synapse cell with large dynamic range can be realized in a compact silicon area. New designs of the synapse cells, neurons, and analog processor are presented. A synapse cell based on Gilbert multiplier structure can perform the linear multiplication for back-propagation networks. A double differential-pair synapse cell can perform the Gaussian function for radial-basis network. The synapse cells can be biased in the strong inversion region for high-speed operation or biased in the subthreshold region for low-power operation. The voltage gain of the sigmoid-function neurons is externally adjustable which greatly facilitates the search of optimal solutions in certain networks. Various building blocks can be intelligently connected to form useful industrial applications. Efficient data communication is a key system-level design issue for large-scale networks. We also present analog neural processors based on perceptron architecture and Hopfield network for communication applications. Biologically inspired neural networks have played an important role towards the creation of powerful intelligent machines. Accuracy, limitations, and prospects of analog current-mode design of the biologically inspired vision processing chips and cellular neural network chips are key design issues.

Sheu, Bing J.↗

DOE OSTI2021

Spray and combustion characteristics of pure hydrothermal liquefaction biofuel and mixture blends with diesel

Hydrothermal liquefaction (HTL) is the process in which biological compounds can be converted into biocrude diesel; various feedstocks, such as algae and wet biowaste, can be utilized. Almost 20 million dry tons of wet biowaste is deposited in U.S. municipal wastewaters annually. If recovered and converted into biodiesel, it can act as a massive source of renewable energy. Preliminary experiments on a diesel engine show that HTL fuel can achieve similar performance and emission levels to diesel. In order to further understand the influence of the HTL fuel properties, the HTL fuel and blends with diesel are observed in a constant volume chamber. Previously, 10% and 20% of the HTL fuel were blended with diesel by volume and demonstrated properties that roughly matched diesel in terms of combustion. In this study, 50% and 100% HTL fuel blends are tested on the same apparatus. It was discovered that the workability of HTL is almost the same as that of diesel and is less sensitive to ambient temperatures. When the ambient temperature is reduced from 1200 K to 800 K, the change ratio of ignition delay is only 62% of diesel. By analyzing the combustion process, it can be seen that there is no need to make major modifications to the engine control strategy if HTL fuel is applied to diesel engines. Though, under low temperature and low oxygen conditions, the soot luminosity peak of HTL100 is 3.12 times than that of diesel.

09 BIOMASS FUELS↗

NASA NTRS2005

Frontiers of Life Sciences: The Human Exploration of the Moon and Mars

The rapid development of the productive processes after World War II extended human settlements into new ecological niches. Advances in Life Sciences played a decisive role supporting the establishment of human presence in areas of the planet where human life could have not existed otherwise. The evolution of life support systems, and the fabrication of new materials and technologies has enabled humans to inhabit Polar Regions, ocean surfaces and depths; and to leave Earth and occupy Low Earth Orbit. By the end of the 20 th Century, stations in the Antarctic and Arctic, off shore oil platforms, submarines, and space stations had become the ultimate demonstration of human ability to engineer habitats at Earth extreme environments and outer space. As we enter the 21st Century, the next development of human settlements will occur through the exploration of the Moon, Mars, and beyond. The major risks of space exploration derive from long exposure of humans and other life systems to radiation, microgravity, isolation and confinement, dependence on artificial life support systems, and unknown effects (e.g., altered magnetic fields, ultrahigh vacuum on bacteria, fungi, etc.). Countermeasures will require a complete characterization of human and other biological systems adaptation processes. To sustain life in transit and on the surface of the Moon and Mars will require a balance of spacecraft, cargo, astronaut crews, and the use of in situ resources. Limitations on the number of crewmembers, payloads, and the barrenness of the terrain require a novel design for the capabilities needed in transit and at exploration outpost sites. The planned destinations have resources that may be accessed to produce materials, food, shelter, power, and to provide an environment compatible with successful occupation of longterm exploration sites. Once more, the advancements of Life Sciences will be essential for the design of interplanetary voyages and planetary surface operations. This presentation delineates the role of Life Sciences and its frontiers, especially Cell Science, in the context of human exploration. Life support systems, food production, and medical equipment encompass many of vital aspects related to the new vision for NASA.

North, Regina M.↗

DOE OSTI2024

Cell-Free Gene Expression: Methods and Applications

Cell-free gene expression (CFE) systems empower synthetic biologists to build biological molecules and processes outside of living intact cells. The foundational principle is that precise, complex biomolecular transformations can be conducted in purified enzyme or crude cell lysate systems. This concept circumvents mechanisms that have evolved to facilitate species survival, bypasses limitations on molecular transport across the cell wall, and provides a significant departure from traditional, cell-based processes that rely on microscopic cellular “reactors.” In addition, cell-free systems are inherently distributable through freeze-drying, which allows simple distribution before rehydration at the point-of-use. Furthermore, as cell-free systems are nonliving, they provide built-in safeguards for biocontainment without the constraints attendant on genetically modified organisms. These features have led to a significant increase in the development and use of CFE systems over the past two decades. Here, we discuss recent advances in CFE systems and highlight how they are transforming efforts to build cells, control genetic networks, and manufacture biobased products.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Insights into the unique carboxylation reactions in the metabolism of propylene and acetone

Alkenes and ketones are two classes of ubiquitous, toxic organic compounds in natural environments produced in several biological and anthropogenic processes. In spite of their toxicity, these compounds are utilized as primary carbon and energy sources or are generated as intermediate metabolites in the metabolism of other compounds by many diverse bacteria. The aerobic metabolism of some of the smallest and most volatile of these compounds (propylene, acetone, isopropanol) involves novel carboxylation reactions resulting in a common product acetoacetate. Propylene is metabolized in a four-step pathway involving five enzymes where the penultimate step is a carboxylation reaction catalyzed by a unique disulfide oxidoreductase that couples reductive cleavage of a thioether linkage with carboxylation to produce acetoacetate. The carboxylation of isopropanol begins with conversion to acetone via an alcohol dehydrogenase. Acetone is converted to acetoacetate in a single step by an acetone carboxylase which couples the hydrolysis of MgATP to the activation of both acetone and bicarbonate, generating highly reactive intermediates that are condensed into acetoacetate at a Mn2+ containing the active site. Acetoacetate is then utilized in central metabolism where it is readily converted to acetyl-coenzyme A and subsequently converted into biomass or utilized in energy metabolism via the tricarboxylic acid cycle. This review summarizes recent structural and biochemical findings that have contributed significant insights into the mechanism of these two unique carboxylating enzymes.

Biochemistry & Molecular Biology↗

NASA NTRS2010

Chiral Biomarkers in Meteorites

The chirality of organic molecules with the asymmetric location of group radicals was discovered in 1848 by Louis Pasteur during his investigations of the rotation of the plane of polarization of light by crystals of sodium ammonium paratartrate. It is well established that the amino acids in proteins are exclusively Levorotary (L-aminos) and the sugars in DNA and RNA are Dextrorotary (D-sugars). This phenomenon of homochirality of biological polymers is a fundamental property of all life known on Earth. Furthermore, abiotic production mechanisms typically yield recemic mixtures (i.e. equal amounts of the two enantiomers). When amino acids were first detected in carbonaceous meteorites, it was concluded that they were racemates. This conclusion was taken as evidence that they were extraterrestrial and produced by abiologically. Subsequent studies by numerous researchers have revealed that many of the amino acids in carbonaceous meteorites exhibit a significant L-excess. The observed chirality is much greater than that produced by any currently known abiotic processes (e.g. Linearly polarized light from neutron stars; Circularly polarized ultraviolet light from faint stars; optically active quartz powders; inclusion polymerization in clay minerals; Vester-Ulbricht hypothesis of parity violations, etc.). This paper compares the measured chirality detected in the amino acids of carbonaceous meteorites with the effect of these diverse abiotic processes. IT is concluded that the levels observed are inconsistent with post-arrival biological contamination or with any of the currently known abiotic production mechanisms. However, they are consistent with ancient biological processes on the meteorite parent body. This paper will consider these chiral biomarkers in view of the detection of possible microfossils found in the Orgueil and Murchison carbonaceous meteorites. Energy dispersive x-ray spectroscopy (EDS) data obtained on these morphological biomarkers will be presented to show that the elemental compositions of these morphological biomarkers are not consistent with the compositions expected for modern (i.e. post-arrival) biological contaminants.

Hoover, Richard B.↗

DOE OSTI2024

Fingerprinting Interactions between Proteins and Ligands for Facilitating Machine Learning in Drug Discovery

Molecular recognition is fundamental in biology, underpinning intricate processes through specific protein–ligand interactions. This understanding is pivotal in drug discovery, yet traditional experimental methods face limitations in exploring the vast chemical space. Computational approaches, notably quantitative structure–activity/property relationship analysis, have gained prominence. Molecular fingerprints encode molecular structures and serve as property profiles, which are essential in drug discovery. While two-dimensional (2D) fingerprints are commonly used, three-dimensional (3D) structural interaction fingerprints offer enhanced structural features specific to target proteins. Machine learning models trained on interaction fingerprints enable precise binding prediction. Recent focus has shifted to structure-based predictive modeling, with machine-learning scoring functions excelling due to feature engineering guided by key interactions. Notably, 3D interaction fingerprints are gaining ground due to their robustness. Various structural interaction fingerprints have been developed and used in drug discovery, each with unique capabilities. This review recapitulates the developed structural interaction fingerprints and provides two case studies to illustrate the power of interaction fingerprint-driven machine learning. The first elucidates structure–activity relationships in β2 adrenoceptor ligands, demonstrating the ability to differentiate agonists and antagonists. The second employs a retrosynthesis-based pre-trained molecular representation to predict protein–ligand dissociation rates, offering insights into binding kinetics. Despite remarkable progress, challenges persist in interpreting complex machine learning models built on 3D fingerprints, emphasizing the need for strategies to make predictions interpretable. Binding site plasticity and induced fit effects pose additional complexities. Interaction fingerprints are promising but require continued research to harness their full potential.

3D structural interaction fingerprints↗

DOE OSTI2022

Similarity of the turbulent kinetic energy dissipation rate distribution in the upper mixed layer of the tropical Indian Ocean

Turbulence within the upper ocean mixed layer plays a key role in various physical, biological, and chemical processes. Between September and November 2011, a dataset of 570 vertical profiles of the turbulent kinetic energy (TKE) dissipation rate, as well as conventional hydrological and meteorological data, were collected in the upper layer of the tropical Indian Ocean. These data were used to statistically analyze the vertical distribution of the TKE dissipation rate in the mixed layer. The arithmetic-mean method made the statistical TKE dissipation rate profile more scattered than the median and geometric-mean methods. The statistical TKE dissipation rate were respectively scaled by the surface buoyancy flux and the TKE dissipation rate at the mixed-layer base. It was found that the TKE dissipation rate scaled by that at the mixed-layer base exhibited better similarity characteristics than that scaled by the surface buoyancy flux, whether the stability parameter D/|L MO | was greater or less than 10, indicating that the TKE dissipation rate at the mixed-layer base is a better characteristic scaling parameter for reflecting the intrinsic structure of the TKE dissipation rate in the mixed layer, where D and L MO are respectively the mixed-layer thickness and the Monin-Obukhov length scale. The parameterization of the TKE dissipation rate at the mixed-layer base on the shear-driven dissipation rate and the surface buoyancy flux was further explored. It was found that the TKE dissipation rate at the mixed-layer base could be well fitted by a linear combination of three terms: the wind-shear-driven dissipation rate, the surface buoyancy flux, and a simple nonlinear coupling term of these two.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2017

Mass Spectrometry-based Assay for High Throughput and High Sensitivity Biomarker Verification

Searching for disease specific biomarkers has become a major undertaking in the biomedical research field as the effective diagnosis, prognosis and treatment of many complex human diseases are largely determined by the availability and the quality of the biomarkers. A successful biomarker as an indicator to a specific biological or pathological process is usually selected from a large group of candidates by a strict verification and validation process. To be clinically useful, the validated biomarkers must be detectable and quantifiable by the selected testing techniques in their related tissues or body fluids. Due to its easy accessibility, protein biomarkers would ideally be identified in blood plasma or serum. However, most disease related protein biomarkers in blood exist at very low concentrations (<1ng/mL) and are “masked” by many none significant species at orders of magnitude higher concentrations. The extreme requirements of measurement sensitivity, dynamic range and specificity make the method development extremely challenging. The current clinical protein biomarker measurement primarily relies on antibody based immunoassays, such as ELISA. Although the technique is sensitive and highly specific, the development of high quality protein antibody is both expensive and time consuming. The limited capability of assay multiplexing also makes the measurement an extremely low throughput one rendering it impractical when hundreds to thousands potential biomarkers need to be quantitatively measured across multiple samples. Mass spectrometry (MS)-based assays have recently shown to be a viable alternative for high throughput and quantitative candidate protein biomarker verification. Among them, the triple quadrupole MS based assay is the most promising one. When it is coupled with liquid chromatography (LC) separation and electrospray ionization (ESI) source, a triple quadrupole mass spectrometer operating in a special selected reaction monitoring (SRM) mode, also known as multiple reaction monitoring (MRM), is capable of quantitatively measuring hundreds of candidate protein biomarkers from a relevant clinical sample in a single analysis. The specificity, reproducibility and sensitivity could be as good as ELISA. Furthermore, SRM MS can also quantify protein isoforms and post-translational modifications, for which traditional antibody-based immunoassays often don’t exist.

Environmental Molecular Sciences Laboratory↗

DOE OSTI2021

Universal Approach to Rapid Amplified Plasmonic Sensing Using Helix Defect Phase Transition Polymers

Abstract Permittivity sensing is a critically important analytical tool for bioscience, environmental, and industrial applications. The response time for commonly used plasmonic permittivity sensors is fundamentally set by the reaction kinetics of chemically adsorbed analytes. In this work, the proposal is to overcome this limit by combining plasmonic sensors with phase transition materials possessing a rapid amplified electrostatic response such as quadrupole moment induced molecular helix reversal. As a proof‐of‐concept, rapid sensing of CO 2 on a phase transition polytetrafluoroethylene substrate and amplification of permittivity response in plasmonic Fabry–Perot sensor is shown. The demonstrated universal approach holds promise for a wide range of applications in fast, real time sensing and monitoring of biological and environmental processes.

Shugayev, Roman↗

DOE OSTI2013

Production facilities for second generation biofuels in the USA and the EU – current status and future perspectives

Second generation biofuel production facilities have been slower to reach large-scale production than was anticipated a few years ago even though in Europe, the Renewable Energy Directive has incentives for, and in the United States there are also financial incentives and, the Energy Independence and Security Act of 2007 has targets for second generation biofuels. There have been a number of facilities that have been cancelled, but starting in 2013 it appears that significant quantities of second generation biofuels will be produced. A variety of conversion processes, thermal and biological, as well as hybrids of the two is being utilized. There will be a variety of fuels ethanol, drop-in fuels (e.g. gasoline, diesel), biodiesel, steam, electricity, bio-oil, sugars, and chemicals; and a variety of feedstocks crop residues, wood, wood wastes, energy crops, waste oils and municipal solid waste (MSW). One approach to reducing the risk of moving from first to second generation biofuel production has been to take incremental steps such as converting the cellulosic part of grains into ethanol in addition to the starch portion. Many of the second generation biofuel facilities are co-located with first generation biofuel production facilities to share infrastructure as well as trade by-products (e.g. excess steam). One of the challenges has been financing, but both private and government sources are being utilized. Private sources include internal corporate funds and debt offerings, and venture capital. Furthermore, government sources include the U.S. federal government, the European Union, European national governments, and state and local governments.

09 BIOMASS FUELS↗

DOE OSTI2019

Bxy‐fuca encoding α‐L‐fucosidase plays crucial roles in development and reproduction of the pathogenic pinewood nematode, Bursaphelenchus xylophilus

Abstract BACKGROUND The pine wood nematode (PWN) Bursaphelenchus xylophilus is the causal agent of pine wilt disease (PWD). This disease is a serious threat to pine forests globally. The fuca gene encodes α‐L‐fucosidase, which plays crucial roles in numerous biological and pathological processes in bacteria, fungi, plants and animals. To find promising control strategies against PWD, we investigated the expression and functions of Bxy‐fuca in B. xylophilus . RESULTS Bxy‐fuca encoding α‐L‐fucosidase is highly conserved within the deduced functional domains and key residues. It is expressed continuously across all developmental stages of B. xylophilus . mRNA in situ hybridization demonstrated that Bxy‐fuca was mainly localized in the body wall muscles and intestine. RNA interference indicated that the zygotic expression of Bxy‐fuca was indispensable for embryogenesis. The rate of B. xylophilus egg hatch was significantly decreased after Bxy‐fuca was interfered. Postembryonic silence of Bxy‐fuca resulted in a dramatic decrease in the longevity of and the total number of eggs produced by B. xylophilus . In addition, the motility of the nematode was greatly hampered with a significant drop in head thrashing frequency. CONCLUSION Bxy‐fuca plays crucial roles in development, lifespan and reproduction of B. xylophilus . Our results provide promising hints for control of PWD by blocking embryogenesis and ontogenesis, decreasing nematode fecundity, and disrupting the connection between B. xylophilus and its vector beetle by preventing nematode movement into the tracheal system. © 2019 Society of Chemical Industry

Tang, Jia↗