Reduction of Hydraulic Conductivity during Inhibition of Exudation from Excised Maize and Barley Roots

The uncoupler, carbonyl cyanide m-chlorophenyl hydrazone (CCCP) is shown to reduce the hydraulic conductivity of barley, maize, mung bean, and onion roots. In barley and maize, the reduction in exudation from excised roots is partly due to the reduction in the permeability of the root to water (I_p), but it can be inferred that the rate of salt release to the xylem, is also inhibited. The action of CCCP on L_p is suggested to be mainly in blocking the symplasmic pathway at the plasmodesmata.     

Relation between permeability to potassium and sodium ions and fusicoccin-stimulated hydrogen-ion efflux in barley roots

Summary Measurements are described of fusicoccin (FC)-stimulated H⁺ efflux in barley (Hordeum vulgare L.) roots when K⁺ and Na⁺ concentrations were varied. In low-salt roots H⁺ efflux was stimulated in both 5 mM KCl and NaCl. In salt-saturated roots H⁺ efflux was stimulated more effectively in KCl than in NaCl solution. The stimulation of H⁺ efflux thus is parallel with the selectivity of these different root preparations for K⁺ and Na⁺ and with estimates of permeability ratios (P _Na/P _K) determined from electrical measurements. It is suggested that the results support electrogenic coupling between FC-stimulated H⁺ efflux and cation uptake.     

Oxidation products of terpenes identified from Dendroctonus and Ips bark beetles

Exposure of adult males and females of Dendroctonus brevicomis and D. frontalis to camphene vapor resulted in oxidation of the terpene to a prominent product, which was identified as 6-hydroxy-camphene (camphenol). Exposure of D. brevicomis adults to myrcene vapor resulted in sex-specific oxidation of the hydrocarbon. A major product in both sexes was identified as 2-methyl-6-methylene-2,7-octadien-1-ol (myrcenol), whereas ipsdienol, a major product in males, was not detected in females. A compound detected in hindguts of feeding males of Ips pini and I. paraconfusus was attributed to the presence of 3-carene in the host (Pinus spp.) and subsequently identified as 1-methyl-5-(α-hydroxy-isopropyl)-cyclohexa-1,3-diene.     

Identification of Bacteriophages for Biocontrol of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae

Pseudomonas syringae pv. actinidiae is a reemerging pathogen which causes bacterial canker of kiwifruit (Actinidia sp.). Since 2008, a global outbreak of P. syringae pv. actinidiae has occurred, and in 2010 this pathogen was detected in New Zealand. The economic impact and the development of resistance in P. syringae pv. actinidiae and other pathovars against antibiotics and copper sprays have led to a search for alternative management strategies. We isolated 275 phages, 258 of which were active against P. syringae pv. actinidiae. Extensive host range testing on P. syringae pv. actinidiae, other pseudomonads, and bacteria isolated from kiwifruit orchards showed that most phages have a narrow host range. Twenty-four were analyzed by electron microscopy, pulse-field gel electrophoresis, and restriction digestion. Their suitability for biocontrol was tested by assessing stability and the absence of lysogeny and transduction. A detailed host range was performed, phage-resistant bacteria were isolated, and resistance to other phages was examined. The phages belonged to the Caudovirales and were analyzed based on morphology and genome size, which showed them to be representatives of Myoviridae, Podoviridae, and Siphoviridae. Twenty-one Myoviridae members have similar morphologies and genome sizes yet differ in restriction patterns, host range, and resistance, indicating a closely related group. Nine of these Myoviridae members were sequenced, and each was unique. The most closely related sequenced phages were a group infecting Pseudomonas aeruginosa and characterized by phages JG004 and PAK_P1. In summary, this study reports the isolation and characterization of P. syringae pv. actinidiae phages and provides a framework for the intelligent formulation of phage biocontrol agents against kiwifruit bacterial canker.     

Global fold of human cannabinoid type 2 receptor probed by solid‐state 13C‐, 15N‐MAS NMR and molecular dynamics simulations

The global fold of human cannabinoid type 2 (CB₂) receptor in the agonist‐bound active state in lipid bilayers was investigated by solid‐state ¹³C‐ and ¹⁵N magic‐angle spinning (MAS) NMR, in combination with chemical‐shift prediction from a structural model of the receptor obtained by microsecond‐long molecular dynamics (MD) simulations. Uniformly ¹³C‐ and ¹⁵N‐labeled CB₂ receptor was expressed in milligram quantities by bacterial fermentation, purified, and functionally reconstituted into liposomes. ¹³C MAS NMR spectra were recorded without sensitivity enhancement for direct comparison of C_α, C_β, and C?O bands of superimposed resonances with predictions from protein structures generated by MD. The experimental NMR spectra matched the calculated spectra reasonably well indicating agreement of the global fold of the protein between experiment and simulations. In particular, the ¹³C chemical shift distribution of C_α resonances was shown to be very sensitive to both the primary amino acid sequence and the secondary structure of CB₂. Thus the shape of the C_α band can be used as an indicator of CB₂ global fold. The prediction from MD simulations indicated that upon receptor activation a rather limited number of amino acid residues, mainly located in the extracellular Loop 2 and the second half of intracellular Loop 3, change their chemical shifts significantly (≥1.5 ppm for carbons and ≥5.0 ppm for nitrogens). Simulated two‐dimensional ¹³C_α(i)? ¹³C?O(i) and ¹³C?O(i)? ¹⁵NH(i + 1) dipolar‐interaction correlation spectra provide guidance for selective amino acid labeling and signal assignment schemes to study the molecular mechanism of activation of CB₂ by solid‐state MAS NMR. Proteins 2014; 82:452–465. © 2013 Wiley Periodicals, Inc.     

Concerted Interconversion between Ionic Lock Substates of the β2 Adrenergic Receptor Revealed by Microsecond Timescale Molecular Dynamics

The recently solved crystallographic structures for the A_2A adenosine receptor and the β₁ and β₂ adrenergic receptors have shown important differences between members of the class-A G-protein-coupled receptors and their archetypal model, rhodopsin, such as the apparent breaking of the ionic lock that stabilizes the inactive structure. Here, we characterize a 1.02 μs all-atom simulation of an apo-β₂ adrenergic receptor that is missing the third intracellular loop to better understand the inactive structure. Although we find that the structure is remarkably rigid, there is a rapid influx of water into the core of the protein, as well as a slight expansion of the molecule relative to the crystal structure. In contrast to the x-ray crystal structures, the ionic lock rapidly reforms, although we see an activation-precursor-like event wherein the ionic lock opens for ∼200 ns, accompanied by movements in the transmembrane helices associated with activation. When the lock reforms, we see the structure return to its inactive conformation. We also find that the ionic lock exists in three states: closed (or locked), semi-open with a bridging water molecule, and open. The interconversion of these states involves the concerted motion of the entire protein. We characterize these states and the concerted motion underlying their interconversion. These findings may help elucidate the connection between key local events and the associated global structural changes during activation.     

Identifying areas at risk of drought‐induced tree mortality across South‐Eastern Australia

South‐East Australia has recently been subjected to two of the worst droughts in the historical record (Millennium Drought, 2000–2009 and Big Dry, 2017–2019). Unfortunately, a lack of forest monitoring has made it difficult to determine whether widespread tree mortality has resulted from these droughts. Anecdotal observations suggest the Big Dry may have led to more significant tree mortality than the Millennium drought. Critically, to be able to robustly project future expected climate change effects on Australian vegetation, we need to assess the vulnerability of Australian trees to drought. Here we implemented a model of plant hydraulics into the Community Atmosphere Biosphere Land Exchange (CABLE) land surface model. We parameterized the drought response behaviour of five broad vegetation types, based on a common garden dry‐down experiment with species originating across a rainfall gradient (188–1,125 mm/year) across South‐East Australia. The new hydraulics model significantly improved (~35%–45% reduction in root mean square error) CABLE’s previous predictions of latent heat fluxes during periods of water stress at two eddy covariance sites in Australia. Landscape‐scale predictions of the greatest percentage loss of hydraulic conductivity (PLC) of about 40%–60%, were broadly consistent with satellite estimates of regions of the greatest change in both droughts. In neither drought did CABLE predict that trees would have reached critical PLC in widespread areas (i.e. it projected a low mortality risk), although the model highlighted critical levels near the desert regions of South‐East Australia where few trees live. Overall, our experimentally constrained model results imply significant resilience to drought conferred by hydraulic function, but also highlight critical data and scientific gaps. Our approach presents a promising avenue to integrate experimental data and make regional‐scale predictions of potential drought‐induced hydraulic failure.     

Using Grazing to Manage Herbaceous Structure for a Heterogeneity-Dependent Bird

Grazing management recommendations often sacrifice the intrinsic heterogeneity of grasslands by prescribing uniform grazing distributions through smaller pastures, increased stocking densities, and reduced grazing periods. The lack of patch-burn grazing in semi-arid landscapes of the western Great Plains in North America requires alternative grazing management strategies to create and maintain heterogeneity of habitat structure (e.g., animal unit distribution, pasture configuration), but knowledge of their effects on grassland fauna is limited. The lesser prairie-chicken (Tympanuchus pallidicinctus), an imperiled, grassland-obligate, native to the southern Great Plains, is an excellent candidate for investigating effects of heterogeneity-based grazing management strategies because it requires diverse microhabitats among life-history stages in a semi-arid landscape. We evaluated influences of heterogeneity-based grazing management strategies on vegetation structure, habitat selection, and nest and adult survival of lesser prairie-chickens in western Kansas, USA. We captured and monitored 116 female lesser prairie-chickens marked with very high frequency (VHF) or global positioning system (GPS) transmitters and collected landscape-scale vegetation and grazing data during 2013–2015. Vegetation structure heterogeneity increased at stocking densities ≤0.26 animal units/ha, where use by nonbreeding female lesser prairie-chickens also increased. Probability of use for nonbreeding lesser prairie-chickens peaked at values of cattle forage use values near 37% and steadily decreased with use ≥40%. Probability of use was positively affected by increasing pasture area. A quadratic relationship existed between growing season deferment and probability of use. We found that 70% of nests were located in grazing units in which grazing pressure was <0.8 animal unit months/ha. Daily nest survival was negatively correlated with grazing pressure. We found no relationship between adult survival and grazing management strategies. Conservation in grasslands expressing flora community composition appropriate for lesser prairie-chickens can maintain appropriate habitat structure heterogeneity through the use of low to moderate stocking densities (<0.26 animal units/ha), greater pasture areas, and site-appropriate deferment periods. Alternative grazing management strategies (e.g., rest-rotation, season-long rest) may be appropriate in grasslands requiring greater heterogeneity or during intensive drought. Grazing management favoring habitat heterogeneity instead of uniform grazing distributions will likely be more conducive for preserving lesser prairie-chicken populations and grassland biodiversity. © 2021 The Wildlife Society.