Comparative analysis of plant immune receptor architectures uncovers host proteins likely targeted by pathogens.

BACKGROUND: Plants deploy immune receptors to detect pathogen-derived molecules and initiate defense responses. Intracellular plant immune receptors called nucleotide-binding leucine-rich repeat (NLR) proteins contain a central nucleotide-binding (NB) domain followed by a series of leucine-rich repeats (LRRs), and are key initiators of plant defense responses. However, recent studies demonstrated that NLRs with non-canonical domain architectures play an important role in plant immunity. These composite immune receptors are thought to arise from fusions between NLRs and additional domains that serve as "baits" for the pathogen-derived effector proteins, thus enabling pathogen recognition. Several names have been proposed to describe these proteins, including "integrated decoys" and "integrated sensors". We adopt and argue for "integrated domains" or NLR-IDs, which describes the product of the fusion without assigning a universal mode of action. RESULTS: We have scanned available plant genome sequences for the full spectrum of NLR-IDs to evaluate the diversity of integrations of potential sensor/decoy domains across flowering plants, including 19 crop species. We manually curated wheat and brassicas and experimentally validated a subset of NLR-IDs in wild and cultivated wheat varieties. We have examined NLR fusions that occur in multiple plant families and identified that some domains show re-occurring integration across lineages. Domains fused to NLRs overlap with previously identified pathogen targets confirming that they act as baits for the pathogen. While some of the integrated domains have been previously implicated in disease resistance, others provide new targets for engineering durable resistance to plant pathogens. CONCLUSIONS: We have built a robust reproducible pipeline for detecting variable domain architectures in plant immune receptors across species. We hypothesize that NLR-IDs that we revealed provide clues to the host proteins targeted by pathogens, and that this information can be deployed to discover new sources of disease resistance

Assessment of air flow distribution and hazardous release dispersion around a single obstacle using Reynolds-averaged Navier-Stokes equations

The flow around a cubical building, with a pollution source at the central point of the top of the cube, is studied. The Reynolds-averaged Navier-Stokes and species concentration equations are solved for Reynolds number, Re = 40,000, is based on the height of the cube. The predictions obtained with the standard, the Kato-Launder, and the low-Reynolds number k-epsilon models are examined with various wall functions for the near wall treatment of the flow. Results are compared against Martinuzzi and Tropea measurements (J. of Fluids Eng., 115, 85–92, 1993) for the flow field and against Li and Meroney (J. of Wind Eng. and Industrial Aerodynamics, 81, 333–345, 1983) experiments and Gaussian models for the concentration distribution. It is found that the present unstructured mesh model performs similarly to the structured mesh models. Results from the Kato-Launder model are closer to the experimental data for the flow patterns and contaminant distribution on the cube's roof. However, the Kato-Launder model has an over-prediction for the recirculation zone and the contaminant distribution windward of the cube. The standard k-epsilon and the low-Reynolds number k-epsilon models predict similar flow patterns and are closer to the experimental data of the cube's windward and side face

Computational assessment of the hazardous release dispersion from a diesel pool fire in a complex building’s area

A hazardous release accident taking place within the complex morphology of an urban setting could cause grave damage both to the population’s safety and to the environment. An unpredicted accident constitutes a complicated physical phenomenon with unanticipated outcomes. This is because, in the event of an unforeseen accident, the dispersion of the hazardous materials exhausted in the environment is determined by unstable parameters such as the wind flow and the complex turbulent diffusion around urban blocks of buildings. Our case study focused on a diesel pool fire accident that occured between an array of nine cubical buildings. The accident was studied with a Large eddy Simulation model based on the Fire Dynamics Simulation method. This model was successfully compared against the nine cubes of the Silsoe experiment. The model’s results were used for the determination of the immediately dangerous to life or health smoke zones of the accident. It was found that the urban geometry defined the hazardous gasses dispersion, thus increasing the toxic mass concentration around the buildings

A Thioredoxin Domain-Containing Protein Interacts with Pepino mosaic virus Triple Gene Block Protein 1

Pepino mosaic virus (PepMV) is a mechanically-transmitted tomato pathogen of importance worldwide. Interactions between the PepMV coat protein and triple gene block protein (TGBp1) with the host heat shock cognate protein 70 and catalase 1 (CAT1), respectively, have been previously reported by our lab. In this study, a novel tomato interactor (SlTXND9) was shown to bind the PepMV TGBp1 in yeast-two-hybrid screening, in vitro pull-down and bimolecular fluorescent complementation (BiFC) assays. SlTXND9 possesses part of the conserved thioredoxin (TRX) active site sequence (W__PC vs. WCXPC), and TXND9 orthologues cluster within the TRX phylogenetic superfamilyclosesttophosducin-likeprotein-3. InPepMV-infectedandhealthyNicotianabenthamiana plants,NbTXND9mRNAlevelswerecomparable,andexpressionlevelsremainedstableinbothlocal and systemic leaves for 10 days post inoculation (dpi), as was also the case for catalase 1 (CAT1). To localize the TXND9 in plant cells, a polyclonal antiserum was produced. Purified α-SlTXND9 immunoglobulin (IgG) consistently detected a set of three protein bands in the range of 27–35 kDa, in the 1000 and 30,000 g pellets, and the soluble fraction of extracts of healthy and PepMV-infected N. benthamiana leaves, but not in the cell wall. These bands likely consist of the homologous protein NbTXND9 and its post-translationally modified derivatives. On electron microscopy, immuno-gold labellingofultrathinsectionsofPepMV-infectedN.benthamianaleavesusingα-SlTXND9IgGrevealed particle accumulation close to plasmodesmata, suggesting a role in virus movement. Taken together, this study highlights a novel tomato-PepMV protein interaction and provides data on its localization in planta. Currently, studies focusing on the biological function of this interaction during PepMV infection are in progress

Large eddy simulation of dispersion of hazardous materials released from a fire accident around a cubical building

The turbulent smoke dispersion from a pool fire around a cubical building is studied using large eddy simulation at a high Reynolds number, corresponding to existing experimental measurements both in laboratory and field test scales. Emphasis of this work is on the smoke dispersion due to two different fuel pool fire accident scenarios, initiated behind the building. For the setup of fire in the first case, crude oil was used with a heat release rate of 7.8 MW, and in the second, diesel oil with a heat release rate of 13.5 MW. It is found that in both fire scenarios, the downstream extent of the toxic zone is approximately the same. This is explained in terms of the fact that the smoke concentration and dispersion are influenced mainly by the convective buoyant forces and the strong turbulence mixing processes within the wake zone of the building. It is suggested that wind is the dominating factor in these accident scenarios, which represent the conditions resulting in the highest toxicity levels

Expression of putative effectors of different Xylella fastidiosa strains triggers cell death‐like responses in various Nicotiana model plants

The wide host range of Xylella fastidiosa (Xf) indicates the existence of yet uncharacterized virulence mechanisms that help pathogens to overcome host defences. Various bioinformatics tools combined with prediction of the functions of putative virulence proteins are valuable approaches to study microbial pathogenicity. We collected a number of putative effectors from three Xf strains belonging to different subspecies: Temecula-1 (subsp. fastidiosa), CoDiRO (subsp. pauca), and Ann-1 (subsp. sandyi). We designed an in planta Agrobacterium-based expression system that drives the expressed proteins to the cell apoplast, in order to investigate their ability to activate defence in Nicotiana model plants. Multiple Xf proteins differentially elicited cell death-like phenotypes in different Nicotiana species. These proteins are members of different enzymatic groups: (a) hydrolases/hydrolase inhibitors, (b) serine proteases, and (c) metal transferases. We also classified the Xf proteins according to their sequential and structural similarities via the I-TASSER online tool. Interestingly, we identified similar proteins that were able to differentially elicit cell death in different cultivars of the same species. Our findings provide a basis for further studies on the mechanisms that underlie both defence activation in Xf resistant hosts and pathogen adaptation in susceptible hosts

Tumuli exploration using surface 3D Electrical Resistivity Tomography

Introduction The direct current resistivity method is nowadays a well established geophysical technique, used routinely and successfully in the detection and mapping of concealed subsurface structures, like walls, ditches and anthropogenic or natural cavities (Dahlin and Zhou, 2004). In archaeological geophysics, tombs constitute the most common subterranean manmade cavities of the greatest archaeological and historical importance. Several successful case studies in the detection of tombs hav..