Mechanisms of Surviving Burial: Dune Grass Interspecific Differences Drive Resource Allocation After Sand Deposition

Sand dunes are important geomorphic formations of coastal ecosystems that are critical in protecting human populations that live in coastal areas. Dune formation is driven by ecomorphodynamic interactions between vegetation and sediment deposition. While there has been extensive research on responses of dune grasses to sand burial, there is a knowledge gap in understanding mechanisms of acclimation between similar, coexistent, dune-building grasses such as Ammophila breviligulata (C3), Spartina patens (C4), and Uniola paniculata (C4). Our goal was to determine how physiological mechanisms of acclimation to sand burial vary between species. We hypothesize that (1) in the presence of burial, resource allocation will be predicated on photosynthetic pathway and that we will be able to characterize the C3 species as a root allocator and the C4 species as leaf allocators. We also hypothesize that (2) despite similarities between these species in habitat, growth form, and life history, leaf, root, and whole plant traits will vary between species when burial is not present. Furthermore, when burial is present, the existing variability in physiological strategy will drive species-specific mechanisms of survival. In a greenhouse experiment, we exposed three dune grass species to different burial treatments: 0 cm (control) and a one-time 25-cm burial to mimic sediment deposition during a storm. At the conclusion of our study, we collected a suite of physiological and morphological functional traits. Results showed that Ammophila decreased allocation to aboveground biomass to maintain root biomass, preserving photosynthesis by allocating nitrogen (N) into light-exposed leaves. Conversely, Uniola and Spartina decreased allocation to belowground production to increase elongation and maintain aboveground biomass. Interestingly, we found that species were functionally distinct when burial was absent; however, all species became more similar when treated with burial. In the presence of burial, species utilized functional traits of rapid growth strategy, although mechanisms of change were interspecifically variable

Lipid Modifications of Sonic Hedgehog Ligand Dictate Cellular Reception and Signal Response

Sonic hedgehog (Shh) signaling regulates cell growth during embryonic development, tissue homeostasis and tumorigenesis. Concentration-dependent cellular responses to secreted Shh protein are essential for tissue patterning. Shh ligand is covalently modified by two lipid moieties, cholesterol and palmitate, and their hydrophobic properties are known to govern the cellular release and formation of soluble multimeric Shh complexes. However, the influences of the lipid moieties on cellular reception and signal response are not well understood.We analyzed fully lipidated Shh and mutant forms to eliminate one or both adducts in NIH3T3 mouse embryonic fibroblasts. Quantitative measurements of recombinant Shh protein concentration, cellular localization, and signaling potency were integrated to determine the contributions of each lipid adduct on ligand cellular localization and signaling potency. We demonstrate that lipid modification is required for cell reception, that either adduct is sufficient to confer cellular association, that the cholesterol adduct anchors ligand to the plasma membrane and that the palmitate adduct augments ligand internalization. We further show that signaling potency correlates directly with cellular concentration of Shh ligand.The findings of this study demonstrate that lipid modification of Shh determines cell concentration and potency, revealing complementary functions of hydrophobic modification in morphogen signaling by attenuating cellular release and augmenting reception of Shh protein in target tissues

Improving Cry8Ka toxin activity towards the cotton boll weevil (Anthonomus grandis)

<p>Abstract</p> <p>Background</p> <p>The cotton boll weevil (<it>Anthonomus grandis</it>) is a serious insect-pest in the Americas, particularly in Brazil. The use of chemical or biological insect control is not effective against the cotton boll weevil because of its endophytic life style. Therefore, the use of biotechnological tools to produce insect-resistant transgenic plants represents an important strategy to reduce the damage to cotton plants caused by the boll weevil. The present study focuses on the identification of novel molecules that show improved toxicity against the cotton boll weevil. <it>In vitro </it>directed molecular evolution through DNA shuffling and phage display screening was applied to enhance the insecticidal activity of variants of the Cry8Ka1 protein of <it>Bacillus thuringiensis</it>.</p> <p>Results</p> <p>Bioassays carried out with <it>A. grandis </it>larvae revealed that the LC₅₀of the screened mutant Cry8Ka5 toxin was 3.15-fold higher than the wild-type Cry8Ka1 toxin. Homology modelling of Cry8Ka1 and the Cry8Ka5 mutant suggested that both proteins retained the typical three-domain Cry family structure. The mutated residues were located mostly in loops and appeared unlikely to interfere with molecular stability.</p> <p>Conclusions</p> <p>The improved toxicity of the Cry8Ka5 mutant obtained in this study will allow the generation of a transgenic cotton event with improved potential to control <it>A. grandis</it>.</p

Investigating dune-building feedback at the plant level: Insights from a multispecies field experiment

Coastal foredunes provide the first line of defense against rising sea levels and storm surge and for this reason there is increasing interest in understanding and modeling foredune formation and post-storm recovery. However, there is limited observational data available to provide empirical guidance for the development of model parameterizations. To provide guidance for improved representation of dune grass growth in models, we conducted a two-year multi-species transplant experiment on Hog Island, VA, U.S.A. and measured the dependence of plant growth on elevation and distance from the shoreline, as well as the relationship between plant growth and sand accumulation. We tracked total leaf growth (length) and aboveground leaf length and found that Ammophila breviligulata (American beachgrass) and Uniola paniculata (sea oats) grew more than Spartina patens (saltmeadow cordgrass) by a factor of 15% (though not statistically significant) and 45%, respectively. Our results also suggest a range of basal/frontal area ratios (an important model parameter) from 0.5-1 and a strong correlation between transplant growth and total sand deposition for all species at the scale of two years, but not over shorter temporal scales. Distance from the shoreline and elevation had no effect on transplant growth rate but did have an effect on survival. Based on transplant survival, the seaward limit of vegetation at the end of the experiment was approximately 30 m from the MHWL and at an elevation of 1.43 m, corresponding to inundation less than 7.5% of the time according to total water level calculations. Results from this experiment provide evidence for the dune-building capacity of all three species, suggesting S. patens is not a maintainer species, as previously thought, but rather a moderate dune builder even though its growth is less stimulated by sand deposition than A. breviligulata and U. paniculata

Molecular basis of caspase-1 polymerization and its inhibition by a new capping mechanism

Inflammasomes are cytosolic caspase-1-activation complexes that sense intrinsic and extrinsic danger signals, and trigger inflammatory responses and pyroptotic cell death. Homotypic interactions among Pyrin domains and caspase recruitment domains (CARDs) in inflammasome-complex components mediate oligomerization into filamentous assemblies. Several cytosolic proteins consisting of only interaction domains exert inhibitory effects on inflammasome assembly. In this study, we determined the structure of the human caspase-1 CARD domain (caspase-1[superscript CARD]) filament by cryo-electron microscopy and investigated the biophysical properties of two caspase-1-like CARD-only proteins: human inhibitor of CARD (INCA or CARD17) and ICEBERG (CARD18). Our results reveal that INCA caps caspase-1 filaments, thereby exerting potent inhibition with low-nanomolar K[subscript i] on caspase-1[superscript CARD] polymerization in vitro and inflammasome activation in cells. Whereas caspase-1[superscript CARD] uses six complementary surfaces of three types for filament assembly, INCA is defective in two of the six interfaces and thus terminates the caspase-1 filament

Crystal structure analysis of free and substrate-bound 6-hydroxy-L-nicotine oxidase from Arthrobacter nicotinovorans

The pathway for oxidative degradation of nicotine in Arthrobacter nicotinovorans includes two genetically and structurally unrelated flavoenzymes, 6-hydroxy-L-nicotine oxidase (6HLNO) and 6-hydroxy-D-nicotine oxidase, which act with absolute stereospecificity on the L- and D-forms, respectively, of 6-hydroxy-nicotine. We solved the crystal structure of 6HLNO at 1.95 A resolution by combined isomorphous/multiple-wavelength anomalous dispersion phasing. The overall structure of each subunit of the 6HLNO homodimer and the folds of the individual domains are closely similar as in eukaryotic monoamine oxidases. Unexpectedly, a diacylglycerophospholipid molecule was found to be non-covalently bound to each protomer of 6HLNO. The fatty acid chains occupy hydrophobic channels that penetrate deep into the interior of the substrate-binding domain of each subunit. The solvent-exposed glycerophosphate moiety is located at the subunit-subunit interface. We further solved the crystal structure of a complex of dithionite-reduced 6HLNO with the natural substrate 6-hydroxy-L-nicotine at 2.05 A resolution. The location of the substrate in a tight cavity suggests that the binding geometry of this unproductive complex may be closely similar as under oxidizing conditions. The observed orientation of the bound substrate relative to the isoalloxazine ring of the flavin adenine dinucleotide cofactor is suitable for hydride-transfer dehydrogenation at the carbon atom that forms the chiral center of the substrate molecule. A comparison of the substrate-binding modes of 6HLNO and 6-hydroxy-D-nicotine oxidase, based on models of complexes with the D-substrate, suggests an explanation for the stereospecificity of both enzymes. The two enzymes are proposed to orient the enantiomeric substrates in mirror symmetry with respect to the plane of the flavin