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Climate change necessitates increased stress resilience of food crops. We describe four potential solutions, with emphasis on a relatively novel approach aiming at true tolerance of drought rather than improved water-holding capacity of crops, which is a common approach in current breeding and genome editing efforts. Some Angiosperms are known to tolerate loss of 95% of their cellular water, without dying, not dissimilar to seeds. The molecular mechanisms and their regulation underlying this remarkable ability are potentially useful to design tolerant crops. Since most crops produce desiccation tolerant seeds, genomic information for this attribute is present but inactive in vegetative parts of the plant. Based on recent evidence from both seeds and desiccation tolerant Angiosperms we address possible routes to 'flipping the switch' to vegetative desiccation tolerance in major crops.

AMPA receptors are tetrameric glutamate‐gated ion channels that mediate a majority of fast excitatory neurotransmission in the brain. They exist as calcium‐impermeable (CI‐) and calcium‐permeable (CP‐) subtypes, the latter of which lacks the GluA2 subunit. CP‐AMPARs display an array of distinctive biophysical and pharmacological properties that allow them to be functionally identified. This has revealed that they play crucial roles in diverse forms of central synaptic plasticity. Here we summarise the functional hallmarks of CP‐AMPARs and describe how these are modified by the presence of auxiliary subunits that have emerged as pivotal regulators of AMPARs. A lasting change in the prevalence of GluA2‐containing AMPARs, and hence in the fraction of CP‐AMPARs, is a feature in many maladaptive forms of synaptic plasticity and neurological disorders. These include modifications of glutamatergic transmission induced by inflammatory pain, fear conditioning, cocaine exposure, and anoxia‐induced damage in neurons and glia. Furthermore, defective RNA editing of GluA2 can cause altered expression of CP‐AMPARs and is implicated in motor neuron damage (amyotrophic lateral sclerosis) and the proliferation of cells in malignant gliomas. A number of the players involved in CP‐AMPAR regulation have been identified, providing useful insight into interventions that may prevent the aberrant CP‐AMPAR expression. Furthermore, recent molecular and pharmacological developments, particularly the discovery of TARP subtype‐selective drugs, offer the exciting potential to modify some of the harmful effects of increased CP‐AMPAR prevalence in a brain region‐specific manner., figure legend AMPARs containing GluA2 (red subunits) are Ca2+‐impermeable (CI‐AMPARs). Those that lack GluA2 are Ca2+‐permeable (CP‐AMPARs) and are implicated in diverse forms of synaptic plasticity and disease. Both native CP‐ and CI‐AMPARs contain various auxiliary subunits (shown as yellow, green or turquoise) that affect AMPAR function and play a role in the regulation of relative CP‐/CI‐AMPAR prevalence. Image based on PDB model 6NJM.

Plant secondary metabolites are significant mediators of the interactions of plants with their environment. The desiccation tolerant “resurrection plant” Myrothamnus flabellifolia Welw. is distributed throughout southern Africa and is vicariantly separated between arid Namibia and the comparatively mesic South Africa where it is subjected to contrasting climate regimes. Our earlier study indicated that these populations were metabolomically and phylogenetically distinct, leading us to hypothesise that the metabolic signature of the arid-adapted plants from Namibia was environmentally determined. To test this, we profiled the metabolomes of plants from both regions that had been sampled directly in the field (non-acclimated) with those of plants from the same regions that had been long-term acclimated in a greenhouse (acclimated). Long-term acclimation resulted in a convergence of the metabolic profiles of the plants collected from the different regions. However, while the South African plants demonstrated little difference between their acclimated and non-acclimated metabolomes, the metabolomes of the acclimated Namibian samples were distinct from those of the plants sampled in the field in Namibia. Further scrutiny of the metabolite differences between the acclimated and non-acclimated Namibian samples revealed several metabolites that were significantly more abundant in the field samples, and we propose that these might contribute to survival in this region. To further test the metabolomic response from a temporal perspective, we subjected the acclimated plants to short-term high-temperature treatment in climate chambers. However, this treatment had no significant impact on the metabolomes of the plants. Our findings suggest that long-term, rather than short-term, environmental conditions warrant large-scale reprogramming of the metabolome in M. flabellifolia, and that the climate conditions in the more arid region of the species’ range necessitate the production of particular phenolic metabolites.

Fisheries enforcement relies on visual catch identification and quantification at sea or when landed. Silage (fish dissolved in acid) and fish blocks (block frozen fish) are promising methods for on-board processing and storage of low-value catches. We examined the use of non-destructive sampling and two DNA-based methods, quantitative PCR (qPCR) and metabarcoding, to assess species composition and relative abundance in industrial grade experimental silage and fish blocks. We demonstrate the ability to identify and quantify DNA from fish species in both products. qPCR analysis of small silage samples collected over 21 days detected all target control species. DNA from one species (Atlantic wolffish) was consistently overrepresented while, for three species of gadoids (Atlantic cod, haddock and whiting), the DNA content matched input tissue proportions with high accuracy. qPCR and metabarcoding of fish blocks, sampled as run-off water and exterior swabs, provided consistent species detection, with the highest variance observed in quantification from swab samples. Our analysis shows that DNA-based methods have significant potential as a tool for species identification and quantification of complex on-board-processed seafood products and are readily applicable to taxonomically and morphologically similar fish. There is, however, a need for establishing DNA/weight calibration factors for primary fisheries species.

Vegetative desiccation tolerance, or the ability to survive the loss of ~95% relative water content (RWC), is rare in angiosperms, with these being commonly called resurrection plants. It is a complex multigenic and multi-factorial trait, with its understanding requiring a comprehensive systems biology approach. The aim of the current study was to conduct a label-free proteomic analysis of leaves of the resurrection plant Xerophyta schlechteri in response to desiccation. A targeted metabolomics approach was validated and correlated to the proteomics, contributing the missing link in studies on this species. Three physiological stages were identified: an early response to drying, during which the leaf tissues declined from full turgor to a RWC of ~80–70%, a mid-response in which the RWC declined to 40% and a late response where the tissues declined to 10% RWC. We identified 517 distinct proteins that were differentially expressed, of which 253 proteins were upregulated and 264 were downregulated in response to the three drying stages. Metabolomics analyses, which included monitoring the levels of a selection of phytohormones, amino acids, sugars, sugar alcohols, fatty acids and organic acids in response to dehydration, correlated with some of the proteomic differences, giving insight into the biological processes apparently involved in desiccation tolerance in this species.

Resilience to abiotic stress is associated with a suite of functional traits related to defense and longevity. Stress tolerant plants are generally slow growing with extended leave lifespans and reduced allocation to reproduction. Resurrection plants are ideal systems to test for trade-offs associated with stress tolerance due to their extreme resiliency. While, growth defense trade-offs are well-characterized, few studies have tested for natural variation associated with tolerating the harshest environments. Here, we surveyed a suite of functional traits related to stress tolerance, leaf economics, and reproductive allocation in natural populations of the South African resurrection plant Myrothamnus flabellifolia. We selected three distinct field sites in South Africa ranging from mesic to xeric. Despite considerable environmental variation across the study area, M. flabellifolia plants were extremely and similarly stress tolerant at all sites. However, we detected notable variation in other life history and morphological traits. Plants in more mesic sites were larger, faster growing, and had more inflorescences. In contrast, plants from the most xeric sites appeared to invest more in persistence and defense, with lower growth rates and less reproductive allocation. Together, this suggests that desiccation tolerance is a binary trait in M. flabellifolia with little natural variation, but that other phenotypes are more labile. The trait syndromes exhibited by plants at the different study sites align with general expectations about growth defense tradeoffs associated with the colonization of extreme environments. We show that plants from the least stressful sites are more reproductive and faster growing, whereas plants from the most stressful sites were slower growing and less reproductive. These findings suggest that M. flabellifolia plants are finely tuned to their environment.

Synechococcus picocyanobacteria are ubiquitous and abundant photosynthetic organisms in the marine environment and contribute for an estimated 16% of the ocean net primary productivity. Their light-harvesting complexes, called phycobilisomes (PBS), are composed of a conserved allophycocyanin core from which radiates six to eight rods with variable phycobiliprotein and chromophore content. This variability allows Synechococcus to optimally exploit the wide variety of spectral niches existing in marine ecosystems. Seven distinct pigment types or subtypes have been identified so far in this taxon, based on the phycobiliprotein composition and/or the proportion of the different chromophores in PBS rods. Most genes involved in their biosynthesis and regulation are located in a dedicated genomic region called the PBS rod region. Here, we examined the variability of gene sequences and organization of this genomic region in a large set of sequenced isolates and natural populations of Synechococcus representative of all known pigment types. All regions start with a tRNA-PheGAA and some possess mobile elements including tyrosine recombinases, suggesting that their genomic plasticity relies on a tycheposon-like mechanism. Comparison of the phylogenies obtained for PBS and core genes revealed that the evolutionary history of PBS rod genes differs from the rest of the genome and is characterized by the co-existence of different alleles and frequent allelic exchange. We propose a scenario for the evolution of the different pigment types and highlight the importance of population-scale mechanisms in maintaining a wide diversity of pigment types in different Synechococcus lineages despite multiple speciation events.

SummaryVegetative desiccation tolerance, the ability to survive loss of over 90% of cellular water, is an extremely rare trait in Angiosperms. Xerophyta schlechteri survives such extreme water deficit by entering prolonged quiescence and suppressing drought-induced senescence in most of the leaf area, except the apical tip. Information on the molecular regulation of senescence in such plants is scarce and this is the first study to investigate such regulation in senescing and non-senescing tissues of the same leaf.Genome-wide RNA sequencing enabled comparison of senescent and non-senescent tissues during desiccation and early rehydration, establishment of the water content range in which senescence is initiated and identification of molecular mechanisms employed to bring about cellular death.Senescence-associated genes (XsSAG) specific to this species were identified and two potential regulatory sites were enriched in regions upstream to these XsSAGs, allowing us to create a model of senescence regulation in X. schlechteri based on homology with known Arabidopsis senescence regulators.We hypothesise that desiccation-driven senescence occurs as a result of a convergence of signals around MAPK6 to trigger WRKY-mediated ethylene synthesis and XsSAG expression, not unlike aging and stress-related senescence in Arabidopsis, but at remarkably lower water contents (

The desiccation-tolerant shrub Myrothamnus flabellifolia has colonised a unique and harsh niche that provides little protection from the elements. It has a wide distribution range in southern Africa, occurring across an environmental gradient that is exceptionally arid in the southwest and highly mesic in the northeast. It is also harvested for use in medicinal preparations, both traditionally and commercially. However, the phytochemical variability of plants from different rainfall regions has not been assessed, nor have the intraspecific relationships been evaluated by means of a rigorously tested phylogeny. The aims of the present study were thus (1) to test a phylogenetic hypothesis for intraspecific relationships in M. flabellifolia; (2) to assess, based on the global metabolomic profiles, whether accessions collected from the three different geographic locations in southern Africa across a rainfall gradient can be differentiated, and if this corroborates the phylogenetic signature; and (3) with the aid of multivariate statistical analysis, identify and evaluate the most significant discriminatory metabolites between the three sampled regions that could act as potential barcodes. The results show that the phylogenetic and metabolomic signatures were congruent, and the metabolomic data were better able to discriminate the different populations collected from the three regions. Several potential barcodes for discriminating the material from the three regions are proposed. Quercetin-rhamnoside and 3-O-methylquercetin, both significant antioxidants, were present at significantly higher quantities in the material from the driest region in the west than from the more mesic regions in the south and east, whereas quercetin-3-O-glucuronide was significantly higher in the latter. A naringenin-like compound or arbutin derivative could discriminate the southern samples from the eastern samples, whereas digalloylglucose differentiated the eastern samples from the southern samples. In summary, the findings of this study imply that the origin of the material should be considered when used in medicinal and cosmetic preparations.

Frontal fibrosing alopecia (FFA) is a recently described inflammatory and scarring type of hair loss affecting almost exclusively women. Despite a dramatic recent increase in incidence the aetiopathogenesis of FFA remains unknown. We undertake genome-wide association studies in females from a UK cohort, comprising 844 cases and 3,760 controls, a Spanish cohort of 172 cases and 385 controls, and perform statistical meta-analysis. We observe genome-wide significant association with FFA at four genomic loci: 2p22.2, 6p21.1, 8q24.22 and 15q2.1. Within the 6p21.1 locus, fine-mapping indicates that the association is driven by the HLA-B*07:02 allele. At 2p22.1, we implicate a putative causal missense variant in CYP1B1, encoding the homonymous xenobiotic- and hormone-processing enzyme. Transcriptomic analysis of affected scalp tissue highlights overrepresentation of transcripts encoding components of innate and adaptive immune response pathways. These findings provide insight into disease pathogenesis and characterise FFA as a genetically predisposed immuno-inflammatory disorder driven by HLA-B*07:02., Frontal fibrosing alopecia (FFA) features lichenoid cutaneous inflammation and scarring hair loss. Here, Tziotzios et al. identify four genetic loci associated with FFA by GWAS followed by Bayesian fine-mapping, co-localisation and HLA imputation which highlights HLA-B*07:02 as a risk factor.

Desiccation tolerance is defined as ‘the ability of tissues to survive loss of 95% of cellular water or dehydration to tissue water contents of ≤0.1 g H2O g–1 dry mass (DM)’. This trait is common in reproductive structures such as seeds, but relatively rare in vegetative tissues, occurring in only 330 (0.86%) species of vascular plants (Box 1) (Proctor and Pence, 2002). Among non-tracheophytes, it is present in virtually all lichens (Kranner et al., 2009) and in 210 (1.04%) bryophytes (Wood, 2007). The ability to survive such extreme water loss has invoked much scientific interest which, among other things, has facilitated applications in germplasm conservation via seed storage (Bewley et al., 2013) and, in the case of vegetative desiccation tolerance, towards production of extremely drought-tolerant crops for food security in a hotter dryer future (Hilhorst and Farrant, 2018).

Soil-transmitted helminths (STH) are endemic and widespread across Sub-Saharan Africa. A community wide soil-transmitted helminth (STH) prevalence survey was performed on the island of Bubaque in Guinea-Bissau using both Kato-katz microscopy and qPCR methodology. Predictors of infection and morbidity indicators were identified using multivariable logistic regression, and diagnostic methods were compared using k statistics. Among 396 participants, prevalence of STH by microscopy was 23.2%, hookworm was the only species identified by this method and the mean infection intensity was 312 eggs per gram. qPCR analysis revealed an overall prevalence of any STH infection of 47.3%, with the majority A. duodenale (32.3%), followed by N. americanus (15.01%) and S. stercoralis (13.2%). A. lumbricoides, and T. trichiura infections were negligible, with a prevalence of 0.25% each. Agreement between diagnostic tests was k = 0.22, interpreted as fair agreement, and infection intensity measured by both methods was only minimally correlated (Rs = -0.03). STH infection overall was more common in females and adults aged 31–40. STH infection was associated with open defaecation, low socio-economic status and further distance to a water-source. The prevalence of anaemia (defined as a binary outcome by the WHO standards for age and sex) was 69.1%, and 44.2% of children were malnourished according to WHO child growth standards. Hookworm infection intensity by faecal egg count showed no statistically significant association with age (Rs 0.06) but S. Stercoralis infection intensity by qPCR cycle threshold was higher in pre-school aged children (Rs = 0.30, p-value 0.03) There was no statistically significant association between STH infection and anaemia (OR 1.0 p = 0.8), stunting (OR 1.9, p-value 0.5) and wasting (OR 2.0, p-value 0.2) in children. This study reveals a persistent reservoir of STH infection across the community, with high rates of anaemia and malnutrition, despite high-coverage of mebendazole mass-drug administration in pre-school children. This reflects the need for a new strategy to soil-transmitted helminth control, to reduce infections and ultimately eliminate transmission., Author summary To date, the overall aim of the WHO policy of STH control through periodic mass-drug administration (MDA) has been morbidity control through a reduction in infection intensity in school-aged children. The main limitation of this approach is the rapid re-infection rate, both from the environment in areas with poor sanitation, and the persistent reservoir in adults who are not included in the MDA programme. As a consequence of this limitation, there have been calls to change the overall aim from morbidity control to the elimination of STH infection altogether by breaking the transmission cycle through community-wide deworming. However, there is a dearth of epidemiological data pertaining to adults in relation to STH and in addition, most surveys exclude the helminth Strongyloides stercoralis, due to limitations of the diagnostic methods used. Our study aims to provide up-to-date epidemiological data of STH prevalence, individual and household risk-factors for infection and the association with anaemia and malnutrition in children under 5 years. From 396 participants who provided a stool sample, we found a high prevalence of STH infection, predominantly the hookworm Ancylostoma duodenale, across all age-groups and both sexes. There was a clear association between STH infection and open defaecation in this study, with use of a protected pit latrine protective against infection. There were high rates of anaemia and malnutrition which had no statistically significant association with STH infection, and are likely multifactorial. This study uncovered an ongoing reservoir of STH infection despite high coverage with mebendazole MDA, high rates of morbidity alongside the STH infection and a clear association between STH infection and inadequate sanitation. This reflects the need for further interventional research to assess the optimum strategy to alleviate morbidity and break the STH transmission cycle in this population.

Summary Cortical GABAergic interneurons are generated in large numbers in the ganglionic eminences and migrate into the cerebral cortex during embryogenesis. At early postnatal stages, during neuronal circuit maturation, autonomous and activity-dependent mechanisms operate within the cortex to adjust cell numbers by eliminating naturally occurring neuron excess. Here, we show that when cortical interneurons are generated in aberrantly high numbers—due to a defect in precursor cell proliferation during embryogenesis—extra parvalbumin interneurons persist in the postnatal mouse cortex during critical periods of cortical network maturation. Even though cell numbers are subsequently normalized, behavioral abnormalities remain in adulthood. This suggests that timely clearance of excess cortical interneurons is critical for correct functional maturation of circuits that drive adult behavior., Graphical abstract, Highlights • CUX2 regulates cortical parvalbumin interneuron number generation in the MGE • Excess parvalbumin (PV) interneuron integrates into the early postnatal cortex • Excess PV interneuron is eliminated through PTEN-dependent mechanisms • Behavioral abnormalities persist long after abnormal interneuron excess is cleared, Cortical interneurons are generated in excess during embryogenesis, and numbers are trimmed soon after birth for optimal inhibition. Magno et al. demonstrate that aberrant parvalbumin interneuron excess persists in the postnatal cortex, spanning critical periods of cortical network maturation. Cell numbers are eventually normalized, but behavioral abnormalities remain in adulthood.

Resurrection plants are vascular species able to sustain extreme desiccation in their vegetative tissues. Despite its potential interest, the role of leaf anatomy in CO2 diffusion and photosynthesis under non-stressed conditions has not been explored in these species. Net CO2 assimilation (An) and its underlying diffusive, biochemical, and anatomical determinants were assessed in 10 resurrection species from diverse locations, including ferns, and homoiochlorophyllous and poikilochlorophyllous angiosperms. Data obtained were compared with previously published results in desiccation-sensitive ferns and angiosperms. An in resurrection plants was mostly driven by mesophyll conductance to CO2 (gm) and limited by CO2 diffusion. Resurrection species had a greater cell wall thickness (Tcw) than desiccation-sensitive plants, a feature associated with limited CO2 diffusion in the mesophyll, but also greater chloroplast exposure to intercellular spaces (Sc), which usually leads to higher gm. This combination enabled a higher An per Tcw compared with desiccation-sensitive species. Resurrection species possess unusual anatomical features that could confer stress tolerance (thick cell walls) without compromising the photosynthetic capacity (high chloroplast exposure). This mechanism is particularly successful in resurrection ferns, which display higher photosynthesis than their desiccation-sensitive counterparts.

We have used reverse transcription-PCR coupled with 52- and 32-RACE to isolate a full length INO1 cDNA (1692�bp with an ORF of 1530) from the resurrection plant Xerophyta viscosa Baker. XvINO1 encodes 510 amino acids, with a predicted MW of 56.7kD and contains four sequence motifs that are highly conserved in plant myo-inositol-1-phosphate synthases (MIPS, EC5.5.1.4), the enzyme that catalyses the first step in the formation of myo-inositol (Ino). Northern and western analyses show that the transcript and protein are constitutively present in leaves but their expression increases, temporarily, in response to both accumulative salt stress (~300�mM NaCl) and desiccation (to 5% relative water content). Leaf Ino concentration increases 40-fold during the first 6�h of salt stress, and levels of this and other carbohydrates (galactinol, sucrose, raffinose, stachyose and hexoses) remain elevated relative to control leaves for the duration of salt stress treatment. The timing and pattern of accumulation of these carbohydrates differ under desiccation stress and we propose that they perform different functions in the respective stresses. These are elaborated in discussion of our data.

Desiccation tolerance has evolved recurrently across diverse land plant lineages as an adaptation for survival in regions where seasonal rainfall drives periodic drying of vegetative tissues. Growing interest in this phenomenon has fueled recent physiological, biochemical, and genomic insights into the mechanistic basis of desiccation tolerance. Although, desiccation tolerance is often viewed as binary and monolithic, substantial variation exists in the phenotype and underlying mechanisms across diverse lineages, heterogeneous populations, and throughout the development of individual plants. Most studies have focused on conserved responses in a subset desiccation-tolerant plants under laboratory conditions. Consequently, the variability and natural diversity of desiccation-tolerant phenotypes remains largely uncharacterized. Here, we discuss the natural variation in desiccation tolerance and argue that leveraging this diversity can improve our mechanistic understanding of desiccation tolerance. We summarize information collected from ~600 desiccation-tolerant land plants and discuss the taxonomic distribution and physiology of desiccation responses. We point out the need to quantify natural diversity of desiccation tolerance on three scales: variation across divergent lineages, intraspecific variation across populations, and variation across tissues and life stages of an individual plant. We conclude that this variability should be accounted for in experimental designs and can be leveraged for deeper insights into the intricacies of desiccation tolerance.

Desiccation of plants is often lethal but is tolerated by the majority of seeds and by vegetative tissues of only a small number of land plants. Desiccation tolerance is an ancient trait, lost from vegetative tissues following the appearance of tracheids but reappearing in several lineages when selection pressures favored its evolution. Cells of all desiccation-tolerant plants and seeds must possess a core set of mechanisms to protect them from desiccation- and rehydration-induced damage. This review explores how desiccation generates cell damage and how tolerant cells assuage the complex array of mechanical, structural, metabolic, and chemical stresses and survive.Likewise, the stress of rehydration requires appropriate mitigating cellular responses. We also explore what comparative genomics, both structural and responsive, have added to our understanding of cellular protection mechanisms induced by desiccation, and how vegetative desiccation tolerance circumvents destructive, stress-induced cell senescence.

The bacterial pathogen Pseudomonas aeruginosa causes devastating infections in immunocompromised hosts, including chronic lung infections in cystic fibrosis patients. To combat infection, the host’s immune system produces the antimicrobial oxidants hypochlorous acid (HOCl) and hypothiocyanous acid (HOSCN). Little is known about how P. aeruginosa responds to and survives attack from these oxidants. To address this, we carried out two approaches: a mutant screen and transcriptional study. We identified the P. aeruginosa transcriptional regulator, RclR, which responds specifically to HOCl and HOSCN stress and is essential for protection against both oxidants. We uncovered a link between the P. aeruginosa transcriptional response to these oxidants and physiological processes associated with pathogenicity, including antibiotic resistance and the type 3 secretion system., Pseudomonas aeruginosa is a significant nosocomial pathogen and is associated with lung infections in cystic fibrosis (CF). Once established, P. aeruginosa infections persist and are rarely eradicated despite host immune cells producing antimicrobial oxidants, including hypochlorous acid (HOCl) and hypothiocyanous acid (HOSCN). There is limited knowledge as to how P. aeruginosa senses, responds to, and protects itself against HOCl and HOSCN and the contribution of such responses to its success as a CF pathogen. To investigate the P. aeruginosa response to these oxidants, we screened 707 transposon mutants, with mutations in regulatory genes, for altered growth following HOCl exposure. We identified regulators of antibiotic resistance, methionine biosynthesis, catabolite repression, and PA14_07340, the homologue of the Escherichia coli HOCl-sensor RclR (30% identical), which are required for protection against HOCl. We have shown that RclR (PA14_07340) protects specifically against HOCl and HOSCN stress and responds to both oxidants by upregulating the expression of a putative peroxiredoxin, rclX (PA14_07355). Transcriptional analysis revealed that while there was specificity in the response to HOCl (231 genes upregulated) and HOSCN (105 genes upregulated), there was considerable overlap, with 74 genes upregulated by both oxidants. These included genes encoding the type 3 secretion system, sulfur and taurine transport, and the MexEF-OprN efflux pump. RclR coordinates part of the response to both oxidants, including upregulation of pyocyanin biosynthesis genes, and, in the presence of HOSCN, downregulation of chaperone genes. These data indicate that the P. aeruginosa response to HOCl and HOSCN is multifaceted, with RclR playing an essential role. IMPORTANCE The bacterial pathogen Pseudomonas aeruginosa causes devastating infections in immunocompromised hosts, including chronic lung infections in cystic fibrosis patients. To combat infection, the host’s immune system produces the antimicrobial oxidants hypochlorous acid (HOCl) and hypothiocyanous acid (HOSCN). Little is known about how P. aeruginosa responds to and survives attack from these oxidants. To address this, we carried out two approaches: a mutant screen and transcriptional study. We identified the P. aeruginosa transcriptional regulator, RclR, which responds specifically to HOCl and HOSCN stress and is essential for protection against both oxidants. We uncovered a link between the P. aeruginosa transcriptional response to these oxidants and physiological processes associated with pathogenicity, including antibiotic resistance and the type 3 secretion system.

Resurrection plants are a polyphyletic group of angiosperms which display true desiccation tolerance (DT) - the ability to survive near complete loss of cellular water for extended periods while recovering metabolic competence upon watering. This is achieved by employing tailored protection behaviours depending on the relative state of (de)hydration. Recent work has raised interest in desiccation-associated changes related to tissue destiny in desiccation tolerant vegetative tissues. Homoiochlorophyllous species, such as those in the Craterostigma genus, maintain their photosynthetic apparatus during the dehydration process, employing considerable protection thereof. In the current study, detailed phenotypic characterisation and pulse-amplitude modulated chlorophyll fluorometry were used to identify the critical water contents at which key physiological changes occur in leaves of Craterostigma pumilum and how this relates to desiccation-associated differentiation between Tip and Base tissues of leaf blades. This differentiation appeared to be realised only during early rehydration and after initial water movement through the leaf tissues. This work provides strong evidence for the existence of desiccation-associated tissue differentiation in C.pumilum and, potentially, other homoiochlorophyllous resurrection plants.

Although staple crops do not survive extended periods of drought, their seeds possess desiccation tolerance (DT), as they survive almost complete dehydration (desiccation) during the late maturation phase of development. Resurrection plants are plant species whose seeds and vegetative tissues are desiccation tolerant. Vegetative DT first arose with the transition from aquatic to terrestrial life forms, but it was lost as plants acquired mechanisms for drought resistance. DT was then confined to seeds, spores, and pollen grains. We review evidence suggesting that angiosperm resurrection plants have reactivated the seed DT program in vegetative tissues. Novel omics technologies are providing a better understanding of the changes necessary for this reactivation and may aid in the development of crop varieties that are better able to survive extreme drought conditions.

Late embryogenesis abundant (LEA) proteins are essential to the ability of resurrection plants and orthodox seeds to protect the subcellular milieu against irreversible damage associated with desiccation. In this work, we investigated the structure and function of six LEA proteins expressed during desiccation in the monocot resurrection species Xerophyta schlechteri (XsLEAs). In silico analyses suggested that XsLEAs are hydrophilic proteins with variable intrinsically disordered protein (IDP) properties. Circular dichroism (CD) analysis indicated that these proteins are mostly unstructured in water but acquire secondary structure in hydrophobic solution, suggesting that structural dynamics may play a role in their function in the subcellular environment. The protective property of XsLEAs was demonstrated by their ability to preserve the activity of the enzyme lactate dehydrogenase (LDH) against desiccation, heat and oxidative stress, as well as growth of Escherichia coli upon exposure to osmotic and salt stress. Subcellular localization analysis indicated that XsLEA recombinant proteins are differentially distributed in the cytoplasm, membranes and nucleus of Nicotiana benthamiana leaves. Interestingly, a LEA_1 family protein (XsLEA1-8), showing the highest disorder-to-order propensity and protective ability in vitro and in vivo, was also able to enhance salt and drought stress tolerance in Arabidopsis thaliana. Together, our results suggest that the structural plasticity of XsLEAs is essential for their protective activity to avoid damage of various subcellular components caused by water deficit stress. XsLEA1-8 constitutes a potential model protein for engineering structural stability in vitro and improvement of water-deficit stress tolerance in plants.

Drought-induced senescence is a degenerative process that involves the degradation of cellular metabolites and photosynthetic pigments and uncontrolled dismantling of cellular membranes and organelles. Angiosperm resurrection plants display vegetative desiccation tolerance and avoid drought-induced senescence in most of their tissues. Developmentally older tissues, however, fail to recover during rehydration and ultimately senesce. Comparison of the desiccation-associated responses of older senescent tissues (ST) with non-ST (NST) will allow for understanding of mechanisms promoting senescence in the former and prevention of senescence in the latter. In the monocotyledonous resurrection plant Xerophyta schlechteri (Baker) N.L. Menezes*, leaf tips senesce following desiccation, whereas the rest of the leaf blade survives. We characterized structural and metabolic changes in ST and NST at varying water contents during desiccation and rehydration. Light and transmission electron microscopy was used to follow anatomical and subcellular responses, and metabolic differences were studied using gas chromatography-mass spectrometry and colorimetric metabolite assays. The results show that drying below 35% relative water content (0.7 gH2O/g dry mass) in ST resulted in the initiation of age-related senescence hallmarks and that these tissues continue this process after rehydration. We propose that an age-related desiccation sensitivity occurs in older tissues, in a process metabolically similar to that observed during age-related senescence in Arabidopsis thaliana.

Significance Desiccation tolerance likely evolved independently several times in grasses, providing an ideal comparative system to identify genetic elements controlling this trait. Using comparative genomics, we identified genomic and expression changes distinguishing the desiccation-tolerant grass Eragrostis nindensis from its desiccation-sensitive crop relative Eragrostis tef. We expanded these analyses to include several cereals to identify broadly conserved and divergent patterns during water stress. We found that the distinction between drought and desiccation in grasses is subtle, where genes with essential roles in seed development are broadly expressed under water stress. Thus we propose that seeds and leaves share common sets of coregulated genes of likely ancient origin, with only a few genes uniquely expressed for desiccation tolerance., Grasses are among the most resilient plants, and some can survive prolonged desiccation in semiarid regions with seasonal rainfall. However, the genetic elements that distinguish grasses that are sensitive versus tolerant to extreme drying are largely unknown. Here, we leveraged comparative genomic approaches with the desiccation-tolerant grass Eragrostis nindensis and the related desiccation-sensitive cereal Eragrostis tef to identify changes underlying desiccation tolerance. These analyses were extended across C4 grasses and cereals to identify broader evolutionary conservation and divergence. Across diverse genomic datasets, we identified changes in chromatin architecture, methylation, gene duplications, and expression dynamics related to desiccation in E. nindensis. It was previously hypothesized that transcriptional rewiring of seed desiccation pathways confers vegetative desiccation tolerance. Here, we demonstrate that the majority of seed-dehydration–related genes showed similar expression patterns in leaves of both desiccation-tolerant and -sensitive species. However, we identified a small set of seed-related orthologs with expression specific to desiccation-tolerant species. This supports a broad role for seed-related genes, where many are involved in typical drought responses, with only a small subset of crucial genes specifically induced in desiccation-tolerant plants.

The leaves and twigs of the desiccation-tolerant medicinal shrub Myrothamnus flabellifolia are harvested for use in traditional and commercial teas and cosmetics due to their phenolic properties. The antioxidant and pharmacological value of this plant has been widely confirmed, however, previous studies typically based their findings on material collected from a single region. The existence of phenolic variability between plants from different geographical regions experiencing different rainfall regimes has thus not been sufficiently evaluated. Furthermore, the anthocyanins present in this plant have not been assessed. The present study thus used an untargeted liquid chromatography-tandem-mass spectrometry approach to profile phenolics in M. flabellifolia material collected from three climatically distinct (high, moderate, and low rainfall) regions representing the western, southern, and eastern extent of the species range in southern Africa. Forty-one putative phenolic compounds, primarily flavonoids, were detected, nine of which are anthocyanins. Several of these compounds are previously unknown from M. flabellifolia. Using multivariate statistics, samples from different regions could be distinguished by their phenolic profiles, supporting the existence of regional phenolic variability. This study indicates that significant phenolic variability exists across the range of M. flabellifolia, which should inform both commercial and traditional cultivation and harvesting strategies.

Plants being sessile organisms are well equipped genomically to respond to environmental stressors peculiar to their habitat. Evolution of plants onto land was enabled by the ability to tolerate extreme water loss (desiccation), a feature that has been retained within genomes but not universally expressed in most land plants today. In the majority of higher plants, desiccation tolerance (DT) is expressed only in reproductive tissues (seeds and pollen), but some 135 angiosperms display vegetative DT. Here, we review genome-level responses associated with DT, pointing out common and yet sometimes discrepant features, the latter relating to evolutionary adaptations to particular niches. Understanding DT can lead to the ultimate production of crops with greater tolerance of drought than is currently realized.

The complement-like pathway of the African malaria mosquito Anopheles gambiae provides protection against infection by diverse pathogens. A functional requirement for a core set of proteins during infections by rodent and human malaria parasites, bacteria, and fungi suggests a similar mechanism operates against different pathogens. However, the extent to which the molecular mechanisms are conserved is unknown. In this study we probed the biochemical responses of complement-like pathway to challenge by the Gram-positive bacterium Staphyloccocus aureus. Western blot analysis of the hemolymph revealed that S. aureus challenge activates a TEP1 convertase-like activity and promotes the depletion of the protein SPCLIP1. S. aureus challenge did not lead to an apparent change in the abundance of the LRIM1/APL1C complex compared to challenge by the Gram-negative bacterium, Escherichia coli. Following up on this observation using a panel of LRIM1 and APL1C antibodies, we found that E. coli challenge, but not S. aureus, specifically activates a protease that cleaves the C-terminus of APL1C. Inhibitor studies in vivo and in vitro protease assays suggest that a serine protease is responsible for APL1C cleavage. This study reveals that despite different challenges converging on activation of a TEP1 convertase-like activity, the mosquito complement-like pathway also includes pathogen-specific reactions.

Summary The group of homoiochlorophyllous resurrection plants evolved the unique capability to survive severe drought stress without dismantling the photosynthetic machinery. This implies that they developed efficient strategies to protect the leaves from reactive oxygen species (ROS) generated by photosynthetic side reactions. These strategies, however, are poorly understood. Here, we performed a detailed study of the photosynthetic machinery in the homoiochlorophyllous resurrection plant Craterostigma pumilum during dehydration and upon recovery from desiccation. During dehydration and rehydration, C. pumilum deactivates and activates partial components of the photosynthetic machinery in a specific order, allowing for coordinated shutdown and subsequent reinstatement of photosynthesis. Early responses to dehydration are the closure of stomata and activation of electron transfer to oxygen accompanied by inactivation of the cytochrome b6f complex leading to attenuation of the photosynthetic linear electron flux (LEF). The decline in LEF is paralleled by a gradual increase in cyclic electron transport to maintain ATP production. At low water contents, inactivation and supramolecular reorganization of photosystem II becomes apparent, accompanied by functional detachment of light-harvesting complexes and interrupted access to plastoquinone. This well-ordered sequence of alterations in the photosynthetic thylakoid membranes helps prepare the plant for the desiccated state and minimize ROS production.

Fungal sensitisation is an important factor in severe asthma, not only for allergic bronchopulmonary aspergillosis (ABPA) but also the more recently described severe asthma with fungal sensitisation (SAFS). It is not known whether these diseases are driven by the presence of airway fungal colonisation. We aimed to determine if both SAFS and ABPA were associated with airway isolation of Aspergillus fumigatus and whether the frequency of isolation changed following anti-fungal treatment.Sputum samples were collected from patients with SAFS, ABPA and a control group without fungal sensitisation. We recorded details of antifungal treatment, serum IgE and A. fumigatus specific IgE levels. In a subgroup (n = 9) we recorded serial sputum PCR measurements before and during itraconazole therapy.244 sputum samples were provided by 135 patients, 41(17%) ABPA, 168(69%) SAFS, and 35(14%) controls. Sputum Aspergillus fumigatus PCR was positive in 61 SAFS patients (70%) and 6 ABPA patients (50%) not on anti-fungal treatment at the time of the test, compared to 3 (9%) in controls (χ(2) = 37.90, p0.001). Consequently, 19 patients with SAFS who were taking antifungal treatment (23%) were significantly less likely to be PCR positive than the 61 patients not on treatment (70%) (χ(2) = 36.66, p0.001). All 9 patients assessed serially during therapy had positive sputum PCR pre-treatment and all became negative during itraconazole treatment.We have shown that isolation of fungus from the airway of severe asthma patients with fungal sensitisation is very common, supporting the hypothesis of a mechanistic link between fungal colonisation and sensitisation.

Land salinization is a major constraint for the practice of agriculture in the world. Considering the extent of this phenomenon, the rehabilitation of ecosystems degraded by salinization has become a priority to guarantee food security in semi-arid environments. The mechanical and chemical approaches for rehabilitating salt-affected soils being expensive, an alternative approach is to develop and utilize biological systems utilizing salt-tolerant plant species. Casuarina species are naturally halotolerant, but this tolerance has been shown to be improved when they are inoculated with arbuscular mycorrhizal fungi (AMF) and/or nitrogen-fixing bacteria (Frankia). Furthermore, Casuarina plantations have been proposed to promote the development of plant diversity. Thus, the aim of the current study was to evaluate the impact of a plantation comprising the species Casuarina inoculated with AMF and Frankia on the diversity of the sub-canopy and adjacent vegetation. Work was conducted on a plantation comprising Casurina equisetifolia and C. glauca variously inoculated with Frankia and Rhizophagus fasciculatus prior to field planting. The experimental area of 2500 m2 was divided into randomized blocks and vegetation sampling was conducted below and outside of the Casuarina canopy in 32 m2 plots. A total of 48 samples were taken annually over 3 years, with 24 taken from below the Casuarina canopy and 24 from outside the canopy. The results obtained show that co-inoculation with Frankia and Rhizophagus fasciculatus improves the height and survival rate of both species. After 4&ndash, 5 years, there was greater species diversity and plant biomass in the sub-canopy environment compared with that of the adjacent environments. Our results suggest that inoculation of beneficial microbes can improve growth of Casuarina species and that planting of such species can improve the diversity of herbaceous vegetation in saline environments.

Grébert, Théophile ... et al.-- This article is contribution number 69 of Tara Oceans.-- 10 pages, 4 figures, supporting information http://www.pnas.org/content/suppl/2018/02/09/1717069115.DCSupplemental, Marine Synechococcus cyanobacteria are major contributors to global oceanic primary production and exhibit a unique diversity of photosynthetic pigments, allowing them to exploit a wide range of light niches. However, the relationship between pigment content and niche partitioning has remained largely undetermined due to the lack of a single-genetic marker resolving all pigment types (PTs). Here, we developed and employed a robust method based on three distinct marker genes (cpcBA, mpeBA, and mpeW) to estimate the relative abundance of all known Synechococcus PTs from metagenomes. Analysis of the Tara Oceans dataset allowed us to reveal the global distribution of Synechococcus PTs and to define their environmental niches. Green-light specialists (PT 3a) dominated in warm, green equatorial waters, whereas blue-light specialists (PT 3c) were particularly abundant in oligotrophic areas. Type IV chromatic acclimaters (CA4-A/B), which are able to dynamically modify their light absorption properties to maximally absorb green or blue light, were unexpectedly the most abundant PT in our dataset and predominated at depth and high latitudes. We also identified populations in which CA4 might be nonfunctional due to the lack of specific CA4 genes, notably in warm high-nutrient low-chlorophyll areas. Major ecotypes within clades I–IV and CRD1 were preferentially associated with a particular PT, while others exhibited a wide range of PTs. Altogether, this study provides important insights into the ecology of Synechococcus and highlights the complex interactions between vertical phylogeny, pigmentation, and environmental parameters that shape Synechococcus community structure and evolution, This work was supported by the French “Agence Nationale de la Recherche” Programs SAMOSA (Synechococcus as a model genus for studying adaptation of marine phytoplankton to environmental changes) (Grant ANR-13-ADAP-0010) and France Génomique (Grant ANR-10-INBS-09), the French Government “Investissements d’Avenir” programs World Ocean Bioressources, Biotechnologies and Earth-System Services (OCEANOMICS) (Grant ANR-11-BTBR-0008), the European Union’s Seventh Framework Programs FP7 MicroB3 (Grant Agreement 287589), and Marine Microorganisms: Cultivation Methods for Improving Their Biotechnological Applications (Macumba; Grant Agreement 311975), UK Natural Environment Research Council Grant NE/I00985X/1, and the Spanish Ministry of Science and Innovation Grant MicroOcean PANGENOMICS (GL2011-26848/BOS)

Drought is one of the main threats to agriculture worldwide because most of our staple crops are not very tolerant to dehydration stress. Some 135 species of angiosperms, however, are extremely tolerant of severe drought. These so‐called resurrection plants are desiccation tolerant because they can withstand drying to water contents below 0.1 gram water per gram dry weight and remain viable for extended periods of time, similar to most seeds. Indeed, recent research has shown that mechanisms of desiccation tolerance of different species, be it vegetative tissues or seeds, utilise the same basic ingredients, including anti‐oxidants, chaperone proteins, and specific carbohydrates. However, there is species‐specific behaviour, related to habitat, e.g. in cell wall properties and the manner in which photo‐oxidative damage is prevented upon drying. Crop plants produce desiccation‐tolerant seeds and, hence, possess the genomic information required for desiccation tolerance, but this is exclusively expressed in the seeds. Elucidation of the mechanisms of desiccation tolerance may enable the ‘release’ of this characteristic in the leaves, stems, and roots.

Presynaptic ionotropic glutamate receptors (iGluRs) play important roles in the control of synaptogenesis and neurotransmitter release, yet their regulation is poorly understood. In particular, the contribution of transmembrane auxiliary proteins, which profoundly shape the trafficking and gating of somatodendritic iGluRs, is unknown. Here we examined the influence of transmembrane AMPAR regulatory proteins (TARPs) on presynaptic AMPARs in cerebellar molecular layer interneurons (MLIs). 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a partial agonist at TARP-associated AMPARs, enhanced spontaneous GABA release in wild-type mice but not instargazermice that lack the prototypical TARP stargazin (γ-2). These findings were replicated in mechanically dissociated Purkinje cells with functional adherent synaptic boutons, demonstrating the presynaptic locus of modulation. In dissociated Purkinje cells fromstargazermice, AMPA was able to enhance mIPSC frequency, but only in the presence of the positive allosteric modulator cyclothiazide. Thus, ordinarily, presynaptic AMPARs are unable to enhance spontaneous release without γ-2, which is required predominantly for its effects on channel gating. Presynaptic AMPARs are known to reduce action potential-driven GABA release from MLIs. Although a G-protein-dependent non-ionotropic mechanism has been suggested to underlie this inhibition, paradoxically we found that γ-2, and thus AMPAR gating, was required. Following glutamate spillover from climbing fibers or application of CNQX, evoked GABA release was reduced; instargazermice such effects were markedly attenuated in acute slices and abolished in the dissociated Purkinje cell-nerve bouton preparation. We suggest that γ-2 association, by increasing charge transfer, allows presynaptic AMPARs to depolarize the bouton membrane sufficiently to modulate both phasic and spontaneous release.

Diffuse white matter injury (DWMI), a leading cause of neurodevelopmental disabilities in preterm infants, is characterized by reduced oligodendrocyte formation. NG2-expressing oligodendrocyte precursor cells (NG2 cells) are exposed to various extrinsic regulatory signals, including the neurotransmitter GABA. We investigated GABAergic signaling to cerebellar white matter NG2 cells in a mouse model of DWMI (chronic neonatal hypoxia). We found that hypoxia caused a loss of GABAA receptor-mediated synaptic input to NG2 cells, extensive proliferation of these cells and delayed oligodendrocyte maturation, leading to dysmyelination. Treatment of control mice with a GABAA receptor antagonist or deletion of the chloride-accumulating transporter NKCC1 mimicked the effects of hypoxia. Conversely, blockade of GABA catabolism or GABA uptake reduced NG2 cell numbers and increased the formation of mature oligodendrocytes both in control and hypoxic mice. Our results indicate that GABAergic signaling regulates NG2 cell differentiation and proliferation in vivo, and suggest that its perturbation is a key factor in DWMI.

Aim: The aim of the study was to screen the market classes of common beans popularly grown in Zambia for their polyphenolic phytochemical profiles. Methodology: Three market classes of common bean (red, grey mottled and brown) were investigated for their polyphenolic phytochemical profiles using the High Performance Liquid Chromatography-Photo Diode Array-Electrospray Ionization-Mass Spectrometry (HPLC-PDA-ESIMS). Results: Various phenolic compounds were identified in the Zambian market classes of common beans. Quinic acid, a syringic acid derivative, ferulic acid derivatives, medioresinol, p-coumaric acid, catechin, gallic acid and ferulic acid were identified in all the market classes investigated.

This chapter reviews the possible mechanisms of desiccation tolerance in seeds. It discusses some of the anatomical, biochemical, and biophysical events associated with the onset and loss of desiccation tolerance in orthodox seeds and compare these with developmental aspects of recalcitrant seeds. If cells can survive the immediate stress imposed on the structure of organelles and macromolecules when water is removed, they are often termed desiccation tolerant. For both recalcitrant and orthodox seeds, the relative level of desiccation tolerance changes throughout development so that embryos become more tolerant as they mature and less tolerant as they germinate. For many seeds, the timing of maximum dry matter accumulation and the acquisition of maximum desiccation tolerance of the embryonic axis are within days. Changes in carbohydrates also occur during seed and pollen development and germination. The plant growth regulator abscisic acid appears to play an important role in the development of desiccation tolerance.

Desiccation tolerance is common in seeds and various other organisms, but only a few angiosperm species possess vegetative desiccation tolerance. These 'resurrection species' may serve as ideal models for the ultimate design of crops with enhanced drought tolerance. To understand the molecular and genetic mechanisms enabling vegetative desiccation tolerance, we produced a high-quality whole-genome sequence for the resurrection plant Xerophyta viscosa and assessed transcriptome changes during its dehydration. Data revealed induction of transcripts typically associated with desiccation tolerance in seeds and involvement of orthologues of ABI3 and ABI5, both key regulators of seed maturation. Dehydration resulted in both increased, but predominantly reduced, transcript abundance of genomic 'clusters of desiccation-associated genes' (CoDAGs), reflecting the cessation of growth that allows for the expression of desiccation tolerance. Vegetative desiccation tolerance in X. viscosa was found to be uncoupled from drought-induced senescence. We provide strong support for the hypothesis that vegetative desiccation tolerance arose by redirection of genetic information from desiccation-tolerant seeds.

The orphan crop, Eragrostis tef, was subjected to controlled drought conditions to observe the physiological parameters and proteins changing in response to dehydration stress. Physiological measurements involving electrolyte leakage, chlorophyll fluorescence and ultra-structural analysis showed tef plants tolerated water loss to 50% relative water content (RWC) before adverse effects in leaf tissues were observed. Proteomic analysis using isobaric tag for relative and absolute quantification (iTRAQ) mass spectrometry and appropriate database searching enabled the detection of 5727 proteins, of which 211 proteins, including a number of spliced variants, were found to be differentially regulated with the imposed stress conditions. Validation of the iTRAQ dataset was done with selected stress-related proteins, fructose-bisphosphate aldolase (FBA) and the protective antioxidant proteins, monodehydroascorbate reductase (MDHAR) and peroxidase (POX). Western blot analyses confirmed protein presence and showed increased protein abundance levels during water deficit while enzymatic activity for FBA, MDHAR and POX increased at selected RWC points. Gene ontology (GO)-term enrichment and analysis revealed terms involved in biotic and abiotic stress response, signaling, transport, cellular homeostasis and pentose metabolic processes, to be enriched in tef upregulated proteins, while terms linked to reactive oxygen species (ROS)-producing processes under water-deficit, such as photosynthesis and associated light harvesting reactions, manganese transport and homeostasis, the synthesis of sugars and cell wall catabolism and modification, to be enriched in tef downregulated proteins.

Barrett's metaplasia is the only known morphological precursor to oesophageal adenocarcinoma and is characterized by replacement of stratified squamous epithelium by columnar epithelium. The cell of origin is uncertain and the molecular mechanisms responsible for the change in cellular phenotype are poorly understood. We therefore explored the role of two transcription factors, Cdx2 and HNF4α in the conversion using primary organ cultures. Biopsy samples from cases of human Barrett's metaplasia were analysed for the presence of CDX2 and HNF4α. A new organ culture system for adult murine oesophagus is described. Using this, Cdx2 and HNF4α were ectopically expressed by adenoviral infection. The phenotype following infection was determined by a combination of PCR, immunohistochemical and morphological analyses. We demonstrate the expression of CDX2 and HNF4α in human biopsy samples. Our oesophageal organ culture system expressed markers characteristic of the normal SSQE: p63, K14, K4 and loricrin. Ectopic expression of HNF4α, but not of Cdx2 induced expression of Tff3, villin, K8 and E-cadherin. HNF4α is sufficient to induce a columnar-like phenotype in adult mouse oesophageal epithelium and is present in the human condition. These data suggest that induction of HNF4α is a key early step in the formation of Barrett's metaplasia and are consistent with an origin of Barrett's metaplasia from the oesophageal epithelium.

RATIONALE The arabinoxylans are one of the main components of plant cell walls and are known to play major roles in plant tissues properties depending in particular on their structural features. It has been recently shown that one of the strategies developed by resurrection plants to overcome dehydration is based on cell wall composition. For this purpose, the structural characterization of arabinoxylans from desiccation-tolerant grass Eragrostis nindensis (E. nindensis) was compared with its close relative, the desiccation-sensitive Eragrostis tef (E. tef) in order to further understand mechansism of desiccation tolerance in resurrection plants. METHODS Ion mobility spectrometry coupled to mass spectrometry (IM-MS) in combination with the conventional mass spectrometric approaches, including matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), electrospray ionization multistage tandem mass spectrometry (ESI-MS(n)) and gas chromatography/mass spectrometry (GC/MS), were used to characterize arabinoxylan fragments obtained after endo-xylanase digestion of leave extracts from E. nindensis and E. tef. RESULTS Whole fingerprinting by MALDI-MS analysis showed the presence of various arabinoxylan fragments within leaves of E. nindensis and E. tef. The monosaccharide composition and some linkage information were determined by GC/MS experiments. Information regarding the branching and sequence details was obtained by ESI-MS(n) experiments after sample permethylation. The presence of structural isomeric ions with different collision cross sections was evidenced by IM-MS which could be differentiated using ESI-MS(n). CONCLUSIONS We have shown that an orthogonal approach, and especially IM-MS associated to ESI-MS(n) (n = 2 to 4) and GC/MS allowed characterization of arabinoxylan fragments of E. nindensis and E. tef and revealed the presence of isomeric structures. The same arabinoxylan structures were identified for both species but in different relative abundance. Moreover, this work illustrated that IM-MS can efficiently separate isomeric structures and advantageously complements the conventional mass spectrometric methodologies used for arabinoxylan structural characterization.

Both lung disease and elevation of blood glucose are associated with increased glucose concentration (from 0.4 to ~4.0 mM) in the airway surface liquid (ASL). This perturbation of ASL glucose makes the airway more susceptible to infection by respiratory pathogens. ASL is minute (~1 μl/cm2) and the measurement of glucose concentration in the small volume ASL is extremely difficult. Therefore, we sought to develop a fluorescent biosensor with sufficient sensitivity to determine glucose concentrations in ASL in situ. We coupled a range of environmentally sensitive fluorophores to mutated forms of a glucose/galactose-binding protein (GBP) including H152C and H152C/A213R and determined their equilibrium binding properties. Of these, GBP H152C/A213R–BADAN (Kd 0.86±0.01 mM, Fmax/F0 3.6) was optimal for glucose sensing and in ASL increased fluorescence when basolateral glucose concentration was raised from 1 to 20 mM. Moreover, interpolation of the data showed that the glucose concentration in ASL was increased, with results similar to that using glucose oxidase analysis. The fluorescence of GBP H152C/A213R–BADAN in native ASL from human airway epithelial cultures in situ was significantly increased over time when basolateral glucose was increased from 5 to 20 mM. Overall our data indicate that this GBP is a useful tool to monitor glucose homoeostasis in the lung.

The possible role of photorespiration as a general stress protection mechanism, and in C(4) plant metabolism, is controversial. In particular, the potential involvement of photorespiration in the acquisition of desiccation tolerance in 'resurrection' plants is unknown. An investigation was carried out into whether photorespiration is present in leaves of the C(4) resurrection plant Sporobolus stapfianus Gandoger (Poaceae) and whether it functions as a mechanism of stress resistance in the desiccation-tolerant younger leaves (YL) of this plant. It is shown that the enzymes involved in the photorespiratory pathway maintain their activity until 88% relative water content (RWC) in both YL and desiccation-sensitive older leaves (OL). In subsequent stages of dehydration stress, the enzymatic activity declined similarly in both YL and OL. The content of the phorespiratory metabolite, serine, and ethanolamine, a direct product of serine decarboxylation, is higher in the early stages of dehydration (88% RWC) in OL, suggesting a transiently enhanced photorespiratory activity in these leaves. This was confirmed by simultaneous gas exchange and fluorescence measurements, showing suppression of the electron transport rate in OL exposed to non-photorespiratory conditions (2% O(2)) at 85% RWC. It is concluded that a higher photorespiratory electron transport occurs in desiccation-sensitive OL, and it is therefore proposed that the capacity to scavenge excess electrons through photorespiration does not contribute to protect leaves of the desiccation-tolerant YL of S. stapfianus during the stress.

Development of the embryo of Encephalartos natalensis from a rudimentary meristematic structure approximately 700 μm in length extends over 6 months after the seed is shed from the strobilus. Throughout its development, the embryo remains attached to a long suspensor. Differentiation of the shoot meristem flanked by two cotyledonary protuberances occurs over the first 2 months, during which peripheral tannin channels become apparent. Tannins, apparently elaborated by the endoplasmic reticulum, first accumulate in the large central vacuole and ultimately fill the channel. By the fourth month of development, the root meristem is apparent and procambial tissue forming discrete vascular bundles can be discerned in the elongating cotyledons. Between 4 and 6 months, mucilage ducts differentiate; after 6 months, when the seed becomes germinable, the embryo is characterised by cotyledons far longer than the axis. Shoot and root meristem cells remain ultrastructurally similar throughout embryo ontogeny, containing small vacuoles, many well-differentiated mitochondria and endoplasmic reticulum (ER) profiles, abundant polysomes, plastids containing small starch deposits and Golgi bodies. Unusually, however, Golgi bodies are infrequent in other cells including those elaborating mucilage which is accumulated in distended ER and apparently secreted into the duct lumen directly by ER-derived vesicles. The non-meristematic cells accumulate massive starch deposits to the exclusion of any protein bodies and only very sparse lipid, features which are considered in terms of the prolonged period of embryo development and the high atmospheric oxygen content of the Carboniferous Period, when cycads are suggested to have originated.

Summary Bardet–Biedl syndrome (BBS) is a human genetic disorder with a spectrum of symptoms caused by primary cilium dysfunction. The disease is caused by mutations in one of at least 17 identified genes, of which seven encode subunits of the BBSome, a protein complex required for specific trafficking events to and from the primary cilium. The molecular mechanisms associated with BBSome function remain to be fully elucidated. Here, we generated null and complemented mutants of the BBSome subunit BBS1 in the protozoan parasite, Leishmania. In the absence of BBS1, extracellular parasites have no apparent defects in growth, flagellum assembly, motility or differentiation in vitro but there is accumulation of vacuole-like structures close to the flagellar pocket. Infectivity of these parasites for macrophages in vitro is reduced compared with wild-type controls but the null parasites retain the ability to differentiate to the intracellular amastigote stage. However, infectivity of BBS1 null parasites is severely compromised in a BALB/c mouse footpad model. We hypothesize that the absence of BBS1 in Leishmania leads to defects in specific trafficking events that affect parasite persistence in the host. This is the first report of an association between the BBSome complex and pathogen infectivity.

Xerophyta humilis is a poikilochlorophyllous monocot resurrection plant used as a model to study vegetative desiccation tolerance. Dehydration imposes tension and ultimate loss of integrity of membranes in desiccation sensitive species. We investigated the predominant molecular species of glycerolipids present in root and leaf tissues, using multiple reaction monitoring mass spectrometry, and then analysed changes therein during dehydration and subsequent rehydration of whole plants. The presence of fatty acids with long carbon chains and with odd numbers of carbons were detected and confirmed by gas chromatography. Dehydration of both leaves and roots resulted in an increase in species containing polyunsaturated fatty acids and a decrease in disaturated species. Upon rehydration, lipid saturation was reversed, with this being initiated immediately upon watering in roots but only 12-24 hr later in leaves. Relative levels of species with short-chained odd-numbered saturated fatty acids decreased during dehydration and increased during rehydration, whereas the reverse trend was observed for long-chained fatty acids. X. humilis has a unique lipid composition, this report being one of the few to demonstrate the presence of odd-numbered fatty acids in plant phosphoglycerolipids.

Desiccation tolerance (DT, the ability of certain organisms to survive severe dehydration) was a key trait in the evolution of life in terrestrial environments. Likely, the development of desiccation-tolerant life forms was accompanied by the acquisition of dormancy or a dormancy-like stage as a second powerful adaptation to cope with variations in the terrestrial environment. These naturally stress tolerant life forms may be a good source of genetic information to generate stress tolerant crops to face a future with predicted higher occurrence of drought. By mining for key genes and mechanisms related to DT and dormancy conserved across different species and life forms, unique candidate key genes may be identified. Here we identify several of these putative key genes, shared among multiple organisms, encoding for proteins involved in protection, growth and energy metabolism. Mutating a selection of these genes in the model plant Arabidopsis thaliana resulted in clear DT-, dormancy- and other seed-associated phenotypes, showing the efficiency and power of our approach and paves the way for the development of drought-stress tolerant crops. Our analysis supports a co-evolution of DT and dormancy by shared mechanisms that favour survival and adaptation to ever-changing environments with strong seasonal fluctuations.