A model suite of green algae within the Scenedesmaceae for investigating contrasting desiccation tolerance and morphology

Author Posting. © The Company of Biologists, 2018. This article is posted here by permission of The Company of Biologists for personal use, not for redistribution. The definitive version was published in Journal of Cell Science 131 (2018): jcs212233, doi:10.1242/jcs.212233.Microscopic green algae inhabiting desert microbiotic crusts are remarkably diverse phylogenetically, and many desert lineages have independently evolved from aquatic ancestors. Here we worked with five desert and aquatic species within the family Scenedesmaceae to examine mechanisms that underlie desiccation tolerance and release of unicellular versus multicellular progeny. Live cell staining and time-lapse confocal imaging coupled with transmission electron microscopy established that the desert and aquatic species all divide by multiple (rather than binary) fission, although progeny were unicellular in three species and multicellular (joined in a sheet-like coenobium) in two. During division, Golgi complexes were localized near nuclei, and all species exhibited dynamic rotation of the daughter cell mass within the mother cell wall at cytokinesis. Differential desiccation tolerance across the five species, assessed from photosynthetic efficiency during desiccation/rehydration cycles, was accompanied by differential accumulation of intracellular reactive oxygen species (ROS) detected using a dye sensitive to intracellular ROS. Further comparative investigation will aim to understand the genetic, ultrastructural and physiological characteristics supporting unicellular versus multicellular coenobial morphology, and the ability of representatives in the Scenedesmaceae to colonize ecologically diverse, even extreme, habitats.This work was supported by the National Science Foundation, Division of Integrative Organismal Systems [1355085 to Z.G.C.], an anonymous donor [to Z.G.C.], the Marine Biological Laboratory [to M.B.] and the Environmental and Molecular Sciences Laboratory (EMSL) [48938 to Z.G.C.], a Department of Energy, Office of Science User Facility sponsored by the Office of Biological and Environmental Research, located at Pacific Northwest National Laboratory.2019-04-1

Electron donor-dependent radionuclide reduction and nanoparticle formation by \u3ci\u3eAnaeromyxobacter dehalogenans\u3c/i\u3e strain 2CP-C

Anaeromyxobacter dehalogenans strain 2CP-C reduces U(VI) and Tc(VII) to U(IV)O2(s) (uraninite) and Tc(IV)O2(S) respectively. Kinetic studies with resting cells revealed that U(VI) or Tc(VII) reduction rates using H2 as electron donor exceeded those observed in acetate-amended incubations. The reduction of U(VI) by A. dehalogenans 2CP-C resulted in extracellular accumulation of ~5 nm uraninite nanoparticles in association with a lectin-binding extracellular polymeric substance (EPS). The electron donor did not affect UO2(S) nanoparticle size or association with EPS, but the utilization of acetate as the source of reducing equivalents resulted in distinct UO2(S) nanoparticle aggregates that were ~50 nm in diameter. In contrast, reduction of Tc(VII) by A. dehalogenans 2CP-C cell suspensions produced dense clusters of TcO2 particles, which were localized within the cell periplasm and on the outside of the outer membrane. In addition to direct reduction, A. dehalogenans 2CP-C cell suspensions reduced Tc(VII) indirectly via an Fe(II)-mediated mechanism. Fe(II) produced by strain 2CP-C from either ferrihydrite or Hanford Site sediment rapidly removed 99Tc(VII)O4– from solution

Electron donor-dependent radionuclide reduction and nanoparticle formation by \u3ci\u3eAnaeromyxobacter dehalogenans\u3c/i\u3e strain 2CP-C

Anaeromyxobacter dehalogenans strain 2CP-C reduces U(VI) and Tc(VII) to U(IV)O2(s) (uraninite) and Tc(IV)O2(S) respectively. Kinetic studies with resting cells revealed that U(VI) or Tc(VII) reduction rates using H2 as electron donor exceeded those observed in acetate-amended incubations. The reduction of U(VI) by A. dehalogenans 2CP-C resulted in extracellular accumulation of ~5 nm uraninite nanoparticles in association with a lectin-binding extracellular polymeric substance (EPS). The electron donor did not affect UO2(S) nanoparticle size or association with EPS, but the utilization of acetate as the source of reducing equivalents resulted in distinct UO2(S) nanoparticle aggregates that were ~50 nm in diameter. In contrast, reduction of Tc(VII) by A. dehalogenans 2CP-C cell suspensions produced dense clusters of TcO2 particles, which were localized within the cell periplasm and on the outside of the outer membrane. In addition to direct reduction, A. dehalogenans 2CP-C cell suspensions reduced Tc(VII) indirectly via an Fe(II)-mediated mechanism. Fe(II) produced by strain 2CP-C from either ferrihydrite or Hanford Site sediment rapidly removed 99Tc(VII)O4– from solution

Dichomitus squalens partially tailors its molecular responses to the composition of solid wood

White-rot fungi, such as Dichomitus squalens, degrade all wood components and inhabit mixed-wood forests containing both soft- and hardwood species. In this study, we evaluated how D. squalens responded to the compositional differences in softwood [guaiacyl (G) lignin and higher mannan content] and hardwood [syringyl/guaiacyl (S/G) lignin and higher xylan content] using semi-natural solid cultures. Spruce (softwood) and birch (hardwood) sticks were degraded by D. squalens as measured by oxidation of the lignins using 2D-NMR. The fungal response as measured by transcriptomics, proteomics and enzyme activities showed a partial tailoring to wood composition. Mannanolytic transcripts and proteins were more abundant in spruce cultures, while a proportionally higher xylanolytic activity was detected in birch cultures. Both wood types induced manganese peroxidases to a much higher level than laccases, but higher transcript and protein levels of the manganese peroxidases were observed on the G-lignin rich spruce. Overall, the molecular responses demonstrated a stronger adaptation to the spruce rather than birch composition, possibly because D. squalens is mainly found degrading softwoods in nature, which supports the ability of the solid wood cultures to reflect the natural environment.Peer reviewe

c-Type Cytochrome-Dependent Formation of U(IV) Nanoparticles by Shewanella oneidensis

Modern approaches for bioremediation of radionuclide contaminated environments are based on the ability of microorganisms to effectively catalyze changes in the oxidation states of metals that in turn influence their solubility. Although microbial metal reduction has been identified as an effective means for immobilizing highly-soluble uranium(VI) complexes in situ, the biomolecular mechanisms of U(VI) reduction are not well understood. Here, we show that c-type cytochromes of a dissimilatory metal-reducing bacterium, Shewanella oneidensis MR-1, are essential for the reduction of U(VI) and formation of extracelluar UO (2) nanoparticles. In particular, the outer membrane (OM) decaheme cytochrome MtrC (metal reduction), previously implicated in Mn(IV) and Fe(III) reduction, directly transferred electrons to U(VI). Additionally, deletions of mtrC and/or omcA significantly affected the in vivo U(VI) reduction rate relative to wild-type MR-1. Similar to the wild-type, the mutants accumulated UO (2) nanoparticles extracellularly to high densities in association with an extracellular polymeric substance (EPS). In wild-type cells, this UO (2)-EPS matrix exhibited glycocalyx-like properties and contained multiple elements of the OM, polysaccharide, and heme-containing proteins. Using a novel combination of methods including synchrotron-based X-ray fluorescence microscopy and high-resolution immune-electron microscopy, we demonstrate a close association of the extracellular UO (2) nanoparticles with MtrC and OmcA (outer membrane cytochrome). This is the first study to our knowledge to directly localize the OM-associated cytochromes with EPS, which contains biogenic UO (2) nanoparticles. In the environment, such association of UO (2) nanoparticles with biopolymers may exert a strong influence on subsequent behavior including susceptibility to oxidation by O (2) or transport in soils and sediments

Diversity of Mn oxides produced by Mn(II)-oxidizing fungi

Manganese (Mn) oxides are environmentally abundant, highly reactive mineral phases that mediate the biogeochemical cycling of nutrients, contaminants, carbon, and numerous other elements. Despite the belief that microorganisms (specifically bacteria and fungi) are responsible for the majority of Mn oxide formation in the environment, the impact of microbial species, physiology, and growth stage on Mn oxide formation is largely unresolved. Here, we couple microscopic and spectroscopic techniques to characterize the Mn oxides produced by four different species of Mn(II)-oxidizing Ascomycete fungi (Plectosphaerella cucumerina strain DS2psM2a2, Pyrenochaeta sp. DS3sAY3a, Stagonospora sp. SRC1lsM3a, and Acremonium strictum strain DS1bioAY4a) isolated from acid mine drainage treatment systems in central Pennsylvania. The site of Mn oxide formation varies greatly among the fungi, including deposition on hyphal surfaces, at the base of reproductive structures (e.g., fruiting bodies), and on envisaged extracellular polymers adjacent to the cell. The primary product of Mn(II) oxidation for all species growing under the same chemical and physical conditions is a nanoparticulate, poorly-crystalline hexagonal birnessite-like phase resembling synthetic d-MnO2. The phylogeny and growth conditions (planktonic versus surface-attached) of the fungi, however, impact the conversion of the initial phyllomanganate to more ordered phases, such as todorokite (A. strictum strain DS1bioAY4a) and triclinic birnessite (Stagonospora sp. SRC1lsM3a). Our findings reveal that the species of Mn(II)-oxidizing fungi impacts the size, morphology, and structure of Mn biooxides, which will likely translate to large differences in the reactivity of the Mn oxide phases.Engineering and Applied Science

Transformation of 2-line ferrihydrite to 6-line ferrihydrite under oxic and anoxic conditions

Mineralogical transformations of 2-line ferrihydrite were studied under oxic and Fe3+-reducing conditions to establish the role, if any, of 6-line ferrihydrite (“well” organized ferrihydrite) in the reaction pathway and as a final product. In oxic experiments, concentrated suspensions (0.42 mol/L Fe3+ in 0.1 mol/L NaClO4) of freshly synthesized 2-line ferrihydrite, with and without 3% Ni2+, were aged at an initial pH = 7.2 (unbuffered and unadjusted) and 25°C for more than three years. X-ray diffraction, transmission electron microscopy, and Mössbauer spectroscopy measurements were performed on the solids after different aging periods. The primary mineralogical products observed were 6-line ferrihydrite and goethite, with minor hematite. Aggregation and crystallization of the 2- line ferrihydrite liberated protons and depressed suspension pH, but coprecipitated Ni2+ retarded this process. The joint, interrelated effects of Ni and pH influenced both the extent of conversion of 2- line ferrihydrite and the identity of the major transformation products. Six-line ferrihydrite dominated in the Ni ferrihydrite suspension, whereas goethite dominated in the absence of Ni. Aggregation-induced crystallization of 2-line ferrihydrite particles seemed responsible for 6-line ferrihydrite formation. Mineralogical changes to Ni ferrihydrite under anaerobic conditions were investigated at circumneutral pH using the Fe3+-reducing bacterium Shewanella putrefaciens. Residual 6-line ferrihydrite dominated bioreduced samples that also contained goethite and magnetite. The conversion of 2-line ferrihydrite to 6-line ferrihydrite was considerably more rapid under anaerobic conditions. The sorption of biogenic Fe2+ apparently induced intra-aggregate transformation of 2-line ferrihydrite to 6-line ferrihydrite. Collectively, abiotic and biotic studies indicated that 6-line ferrihydrite can be a transformation product of 2-line ferrihydrite, especially when 2-line ferrihydrite is undergoing transformation to more stable hematite or magnetite