Isotria medeoloides, a North American Threatened Orchid: Fungal Abundance May Be as Important as Light in Species Management.

The management of endangered or threatened plant species is difficult if protocols are not developed to propagate species for the purpose of restoration or the enhancement of existing populations. The management of endangered and threatened orchids is especially difficult because of the obligate interactions between orchids and orchid mycorrhizal fungi. Isotria medeoloides is a federally threatened forest-dwelling orchid species with a wide distribution in eastern North America. Seeds have not been successfully germinated and current management is based primarily on using subcanopy thinning to increase light in areas where monitoring demonstrates that populations are declining. We report the results of long-term monitoring efforts, canopy thinning, and orchid mycorrhizal fungus abundance studies at two locations in Virginia. The declining populations responded positively to the experimental and natural thinning of the canopy. At one site, the response was the result of understory canopy thinning. At the second site, the response was due to the natural death of a canopy tree. In light of the dramatic increase in fungal abundance following death of the canopy tree, we propose the Fungal Abundance Hypothesis as an additional approach to the management of endangered plant species. The removal of canopy trees in or adjacent to Isotria populations results in an increase in dead belowground biomass (i.e., roots of the dead canopy tree) that provides substrates for microbial growth, including orchid mycorrhizal fungi, that benefit Isotria.

Association of Model Neurotransmitters with Lipid Bilayer Membranes.

Aimed at reproducing the results of electrophysiological studies of synaptic signal transduction, conventional models of neurotransmission are based on the specific binding of neurotransmitters to ligand-gated receptor ion channels. However, the complex kinetic behavior observed in synaptic transmission cannot be reproduced in a standard kinetic model without the ad hoc postulation of additional conformational channel states. On the other hand, if one invokes unspecific neurotransmitter adsorption to the bilayer-a process not considered in the established models-the electrophysiological data can be rationalized with only the standard set of three conformational receptor states that also depend on this indirect coupling of neurotransmitters via their membrane interaction. Experimental verification has been difficult because binding affinities of neurotransmitters to the lipid bilayer are low. We quantify this interaction with surface plasmon resonance to measure equilibrium dissociation constants in neurotransmitter membrane association. Neutron reflection measurements on artificial membranes, so-called sparsely tethered bilayer lipid membranes, reveal the structural aspects of neurotransmitters' association with zwitterionic and anionic bilayers. We thus establish that serotonin interacts nonspecifically with the membrane at physiologically relevant concentrations, whereas γ-aminobutyric acid does not. Surface plasmon resonance shows that serotonin adsorbs with millimolar affinity, and neutron reflectometry shows that it penetrates the membrane deeply, whereas γ-aminobutyric is excluded from the bilayer.

In Situ Nanoscale Characterization of Water Penetration through Plasma Polymerized Coatings.

The search continues for means of making quick determinations of the efficacy of a coating for protecting a metal surface against corrosion. One means of reducing the time scale needed to differentiate the performance of different coatings is to draw from nanoscale measurements inferences about macroscopic behavior. Here we connect observations of the penetration of water into plasma polymerized (PP) protective coatings and the character of the interface between the coating and an oxide-coated aluminum substrate or model oxide-coated silicon substrate to the macroscopically observable corrosion for those systems. A plasma polymerized film from hexamethyldisiloxane (HMDSO) monomer is taken as illustrative of a hydrophobic coating, while a PP film from maleic anhydride (MA) is used as a characteristically hydrophilic coating. The neutron reflectivity (NR) of films on silicon oxide coated substrates shows that water moves more readily through the hydrophilic PP-MA film. Off-specular X-ray scattering indicates the PP-MA film on aluminum is less conformal with the substrate than is the PP-HMDSO film. Measurements with infrared-visible sum frequency generation spectroscopy (SFG), which probes the chemical nature of the interface, make clear that the chemical interactions between coating and aluminum oxide are disrupted by interfacial water. With this water penetration and interface disruption, macroscopic corrosion can occur much more rapidly. An Al panel coated with PP-MA corrodes after 1 day in salt spray, while a similarly thin (∼30 nm) PP-HMDSO coating protects an Al panel for a period on the order of one month.

The cytosolic domain of T-cell receptor ζ associates with membranes in a dynamic equilibrium and deeply penetrates the bilayer.

Interactions between lipid bilayers and the membrane-proximal regions of membrane-associated proteins play important roles in regulating membrane protein structure and function. The T-cell antigen receptor is an assembly of eight single-pass membrane-spanning subunits on the surface of T lymphocytes that initiates cytosolic signaling cascades upon binding antigens presented by MHC-family proteins on antigen-presenting cells. Its ζ-subunit contains multiple cytosolic immunoreceptor tyrosine-based activation motifs involved in signal transduction, and this subunit by itself is sufficient to couple extracellular stimuli to intracellular signaling events. Interactions of the cytosolic domain of ζ (ζcyt) with acidic lipids have been implicated in the initiation and regulation of transmembrane signaling. ζcyt is unstructured in solution. Interaction with acidic phospholipids induces structure, but its disposition when bound to lipid bilayers is controversial. Here, using surface plasmon resonance and neutron reflection, we characterized the interaction of ζcyt with planar lipid bilayers containing mixtures of acidic and neutral lipids. We observed two binding modes of ζcyt to the bilayers in dynamic equilibrium: one in which ζcyt is peripherally associated with lipid headgroups and one in which it penetrates deeply into the bilayer. Such an equilibrium between the peripherally bound and embedded forms of ζcyt apparently controls accessibility of the immunoreceptor tyrosine-based activation signal transduction pathway. Our results reconcile conflicting findings of the ζ structure reported in previous studies and provide a framework for understanding how lipid interactions regulate motifs to tyrosine kinases and may regulate the T-cell antigen receptor biological activities for this cell-surface receptor system.