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Salt strengthened the bond formed and quickened the bonding process of 3–5 wt. % hectorite gels during the structural rejuvenation process. This even occurred at 0.002M KCl. Microstructure showed exfoliated, flexible platelet bonding in (+)edge–(−)face configurations. The display of prominent aging time-dependent behavior is due to the structural rejuvenation process being controlled by the electric double layer (EDL) repulsive force. Salt increased the lower energy paths to bonding in the (+)edge–(−)face configurations and weakened the EDL force to form stronger bonds. The Leong model time constant data supported the faster bonding process. In shear, the gels with a weakened EDL repulsive force caused by 0.01 and 0.1M KCl treatment were unable to display EDL force-control time-dependent behavior in the stepdown shear stress response. This situation was remedied by increasing the negative charge density of platelets with adsorbed P2O74−. The amount of P2O74− needed was higher at 0.1M KCl. [ABSTRACT FROM AUTHOR]

Here, we report the frequency-dependent spectrum of ice Ih in the range of 0.2–2 THz. We confirm the presence of a feature that blue-shifts from around 1.55–1.65 THz with a decreasing temperature from 260 to 160 K. There is also a change in the trend of the refractive index of ice corresponding to a dispersion, which is also around 1.6 THz. The features are reproduced in data acquired with three commercial terahertz time-domain spectrometers. Computer-simulated spectra assign the feature to lattice translations perpendicular to the 110 and 1 ̄ 10 planes of the ice Ih crystal. The feature's existence should be recognized in the terahertz measurements of frozen aqueous solution samples to avoid false interpretations. [ABSTRACT FROM AUTHOR]

Background and aims: An overlooked fraction of the terrestrial carbon (C) pool is that associated with biogenic silica deposited in plants (phytoliths), so-called PhytOC. This fraction is small compared with the main C pools, but is of interest because it could be a long-term C sink as phytoliths may protect organic C from mineralization. However, the topic is hotly contested and unclear due to both methodological and theoretical limitations. Scope: We aim to review this topic, with specific emphasis on: (i) the range of C concentrations associated with phytoliths; (ii) soil phytolith preservation and subsequent organic C mineralization; and (iii) global estimates of C sequestration within PhytOC. Conclusions: Recent work has suggested that [PhytOC] could be much greater than currently acknowledged, but also highly variable and dependent on cell silicification types. A short case study using cryo‐Scanning Electron Microscopy (cryo-SEM), X‐ray microanalysis (EDX), plus Focused Ion Beam (FIB) and Scanning Transmission Electron Microscopy (STEM) on the culms of a sedge (Schoenus caespititius) confirmed this thinking. Understanding of both phytolith and PhytOC fates in soil is poor. We suggest that phytolith residence time should be seen as a gradient. Such a continuum is explained by different phytolith sizes, types and chemistry, which will also have contrasting PhytOC. Our estimation of C sequestration as PhytOC each year (11–190 Tg C yr−1) represents between < 1% and 13% of the C that could be sequestered globally in soils (estimated at 1400 Tg C yr−1). We conclude that (1) more research is needed to improve our understanding of the formation and fate of PhytOC in terrestrial ecosystems and (2) it would be unwise to put our faith in PhytOC sequestration or other related methodologies to "solve" the climate crisis. [ABSTRACT FROM AUTHOR]

Salinity tolerance requires coordinated responses encompassing salt exclusion in roots and tissue/cellular compartmentation of salt in leaves. We investigated the possible control points for salt ions transport in roots and tissue tolerance to Na+ and Cl– in leaves of two contrasting mungbean genotypes, salt‐tolerant Jade AU and salt‐sensitive BARI Mung‐6, grown in nonsaline and saline (75 mM NaCl) soil. Cryo‐SEM X‐ray microanalysis was used to determine concentrations of Na, Cl, K, Ca, Mg, P, and S in various cell types in roots related to the development of apoplastic barriers, and in leaves related to photosynthetic performance. Jade AU exhibited superior salt exclusion by accumulating higher [Na] in the inner cortex, endodermis, and pericycle with reduced [Na] in xylem vessels and accumulating [Cl] in cortical cell vacuoles compared to BARI Mung‐6. Jade AU maintained higher [K] in root cells than BARI Mung‐6. In leaves, Jade AU maintained lower [Na] and [Cl] in chloroplasts and preferentially accumulated [K] in mesophyll cells than BARI Mung‐6, resulting in higher photosynthetic efficiency. Salinity tolerance in Jade AU was associated with shoot Na and Cl exclusion, effective regulation of Na and Cl accumulation in chloroplasts, and maintenance of high K in root and leaf mesophyll cells. Summary statement: Salinity tolerance in mungbean is related to its ability to intercept Na and Cl in roots, thereby reducing Na and Cl transport to shoot, effectively regulate Na and Cl accumulation in chloroplasts, and maintain high K in root and leaf mesophyll cells. [ABSTRACT FROM AUTHOR]

Background and aims: Proteaceae are a prominent plant family in south-western Australia. Most Proteaceae are 'calcifuge', occurring exclusively on old phosphorus (P)-impoverished acidic soils, with a few 'soil-indifferent' species also found on young P-richer calcareous soils. Calcium (Ca)-enhanced P toxicity explains the calcifuge habit of Proteaceae. However, previous research has so far been focused exclusively on the roles of Ca and P in determining Proteaceae distribution, and consequently there is little knowledge on how other soil-based strategies influence this distribution. We aimed to study the effects of young calcareous soils on four soil-grown Proteaceae and assess differences between calcifuge and soil-indifferent Proteaceae to better understand their natural distribution. Methods: Two calcifuge and two soil-indifferent Proteaceae from south-western Australia were grown in six contrasting soils, including young calcareous, and old acidic soils. Results: When grown in calcareous soils all species showed root growth inhibition, micronutrient deficiency, Ca-enhanced P toxicity, and negative impacts on physiology. Calcifuge species were more sensitive to calcareous soils than soil-indifferent ones, although this varied between genera. Soil-indifferent species tended to produce more cluster roots, release more carboxylates per root mass, and allocate less Ca to their leaves, compared with calcifuges; they also had smaller seeds and were less sensitive to Ca-enhanced P toxicity. Conclusion: We surmise that a combination of these traits allows soil-indifferent species to tolerate calcareous soils. This study provides insight into how Proteaceae respond to young calcareous soils and how this influences their distribution. [ABSTRACT FROM AUTHOR]

Cryptosporidium is a major cause of diarrhoeal disease and mortality in young children in resource-poor countries, for which no vaccines or adequate therapeutic options are available. Infection in humans is primarily caused by two species: C. hominis and C. parvum. Despite C. hominis being the dominant species infecting humans in most countries, very little is known about its growth characteristics and life cycle in vitro, given that the majority of our knowledge of the in vitro development of Cryptosporidium has been based on C. parvum. In the present study, the growth and development of two C. parvum isolates (subtypes Iowa-IIaA17G2R1 and IIaA18G3R1) and one C. hominis isolate (subtype IdA15G1) in HCT-8 cells were examined and compared at 24 h and 48 h using morphological data acquired with scanning electron microscopy. Our data indicated no significant differences in the proportion of meronts or merozoites between species or subtypes at either time-point. Sexual development was observed at the 48-h time-point across both species through observations of both microgamonts and macrogamonts, with a higher frequency of macrogamont observations in C. hominis (IdA15G1) cultures at 48-h post-infection compared to both C. parvum subtypes. This corresponded to differences in the proportion of trophozoites observed at the same time point. No differences in proportion of microgamonts were observed between the three subtypes, which were rarely observed across all cultures. In summary, our data indicate that asexual development of C. hominis is similar to that of C. parvum, while sexual development is accelerated in C. hominis. This study provides new insights into differences in the in vitro growth characteristics of C. hominis when compared to C. parvum, which will facilitate our understanding of the sexual development of both species. [ABSTRACT FROM AUTHOR]

Although significant intraspecific variation in photosynthetic phosphorus (P) use efficiency (PPUE) has been shown in numerous species, we still know little about the biochemical basis for differences in PPUE among genotypes within a species. Here, we grew two high PPUE and two low PPUE chickpea (Cicer arietinum) genotypes with low P supply in a glasshouse to compare their photosynthesis-related traits, total foliar P concentration ([P]) and chemical P fractions (i.e. inorganic P (Pi), metabolite P, lipid P, nucleic acid P, and residual P). Foliar cell-specific nutrient concentrations including P were characterized using elemental X-ray microanalysis. Genotypes with high PPUE showed lower total foliar [P] without slower photosynthetic rates. No consistent differences in cellular [P] between the epidermis and mesophyll cells occurred across the four genotypes. In contrast, high PPUE was associated with lower allocation to Pi and metabolite P, with PPUE being negatively correlated with the percentage of these two fractions. Furthermore, a lower allocation to Pi and metabolite P was correlated with a greater allocation to nucleic acid P, but not to lipid P. Collectively, our results suggest that a different allocation to foliar P fractions, rather than preferential P allocation to specific leaf tissues, underlies the contrasting PPUE among chickpea genotypes. [ABSTRACT FROM AUTHOR]

Introduction: Biological N2 fixation in feather-mosses is one of the largest inputs of new nitrogen (N) to boreal forest ecosystems; however, revealing the fate of newly fixed N within the bryosphere (i.e. bryophytes and their associated organisms) remains uncertain. Methods: Herein, we combined 15N tracers, high resolution secondary ion mass-spectrometry (NanoSIMS) and a molecular survey of bacterial, fungal and diazotrophic communities, to determine the origin and transfer pathways of newly fixed N2 within feather-moss (Pleurozium schreberi) and its associated microbiome. Results: NanoSIMS images reveal that newly fixed N2, derived from cyanobacteria, is incorporated into moss tissues and associated bacteria, fungi and micro-algae. Discussion: These images demonstrate that previous assumptions that newly fixed N2 is sequestered into moss tissue and only released by decomposition are not correct. We provide the first empirical evidence of new pathways for N2 fixed in feather-mosses to enter the boreal forest ecosystem (i.e. through its microbiome) and discuss the implications for wider ecosystem function. [ABSTRACT FROM AUTHOR]

Parasites are widespread and diverse in oceanic plankton and many of them infect single-celled algae for survival. How these parasites develop and scavenge energy within the host and how the cellular organization and metabolism of the host is altered remain open questions. Combining quantitative structural and chemical imaging with time-resolved transcriptomics, we unveil dramatic morphological and metabolic changes of the marine parasite Amoebophrya (Syndiniales) during intracellular infection, particularly following engulfment and digestion of nutrient-rich host chromosomes. Changes include a sequential acristate and cristate mitochondrion with a 200-fold increase in volume, a 13-fold increase in nucleus volume, development of Golgi apparatus and a metabolic switch from glycolysis (within the host) to TCA (free-living dinospore). Similar changes are seen in apicomplexan parasites, thus underlining convergent traits driven by metabolic constraints and the infection cycle. In the algal host, energy-producing organelles (plastid, mitochondria) remain relatively intact during most of the infection. We also observed that sugar reserves diminish while lipid droplets increase. Rapid infection of the host nucleus could be a "zombifying" strategy, allowing the parasite to digest nutrient-rich chromosomes and escape cytoplasmic defense, whilst benefiting from maintained carbon-energy production of the host cell. [ABSTRACT FROM AUTHOR]

Halophytes accumulate and sequester high concentrations of salt in vacuoles while maintaining lower levels of salt in the cytoplasm. The current data on cellular and subcellular partitioning of salt in halophytes are, however, limited to only a few dicotyledonous C3 species. Using cryo‐scanning electron microscopy X‐ray microanalysis, we assessed the concentrations of Na, Cl, K, Ca, Mg, P and S in various cell types within the leaf‐blades of a monocotyledonous C4 halophyte, Rhodes grass (Chloris gayana). We also linked, for the first time, elemental concentrations in chloroplasts of mesophyll and bundle sheath cells to their ultrastructure and photosynthetic performance of plants grown in nonsaline and saline (200 mM NaCl) conditions. Na and Cl accumulated to the highest levels in xylem parenchyma and epidermal cells, but were maintained at lower concentrations in photosynthetically active mesophyll and bundle sheath cells. Concentrations of Na and Cl in chloroplasts of mesophyll and bundle sheath cells were lower than in their respective vacuoles. No ultrastructural changes were observed in either mesophyll or bundle sheath chloroplasts, and photosynthetic activity was maintained in saline conditions. Salinity tolerance in Rhodes grass is related to specific cellular Na and Cl distributions in leaf tissues, and the ability to regulate Na and Cl concentrations in chloroplasts. [ABSTRACT FROM AUTHOR]

Summary: Ectomycorrhizal plants trade plant‐assimilated carbon for soil nutrients with their fungal partners. The underlying mechanisms, however, are not fully understood. Here we investigate the exchange of carbon for nitrogen in the ectomycorrhizal symbiosis of Fagus sylvatica across different spatial scales from the root system to the cellular level.We provided 15N‐labelled nitrogen to mycorrhizal hyphae associated with one half of the root system of young beech trees, while exposing plants to a 13CO2 atmosphere. We analysed the short‐term distribution of 13C and 15N in the root system with isotope‐ratio mass spectrometry, and at the cellular scale within a mycorrhizal root tip with nanoscale secondary ion mass spectrometry (NanoSIMS).At the root system scale, plants did not allocate more 13C to root parts that received more 15N. Nanoscale secondary ion mass spectrometry imaging, however, revealed a highly heterogenous, and spatially significantly correlated distribution of 13C and 15N at the cellular scale.Our results indicate that, on a coarse scale, plants do not allocate a larger proportion of photoassimilated C to root parts associated with N‐delivering ectomycorrhizal fungi. Within the ectomycorrhizal tissue, however, recently plant‐assimilated C and fungus‐delivered N were spatially strongly coupled. Here, NanoSIMS visualisation provides an initial insight into the regulation of ectomycorrhizal C and N exchange at the microscale. [ABSTRACT FROM AUTHOR]

Background: Sponges are increasingly recognised as key ecosystem engineers in many aquatic habitats. They play an important role in nutrient cycling due to their unrivalled capacity for processing both dissolved and particulate organic matter (DOM and POM) and the exceptional metabolic repertoire of their diverse and abundant microbial communities. Functional studies determining the role of host and microbiome in organic nutrient uptake and exchange, however, are limited. Therefore, we coupled pulse-chase isotopic tracer techniques with nanoscale secondary ion mass spectrometry (NanoSIMS) to visualise the uptake and translocation of 13C- and 15N-labelled dissolved and particulate organic food at subcellular level in the high microbial abundance sponge Plakortis angulospiculatus and the low microbial abundance sponge Halisarca caerulea. Results: The two sponge species showed significant enrichment of DOM- and POM-derived 13C and 15N into their tissue over time. Microbial symbionts were actively involved in the assimilation of DOM, but host filtering cells (choanocytes) appeared to be the primary site of DOM and POM uptake in both sponge species overall, via pinocytosis and phagocytosis, respectively. Translocation of carbon and nitrogen from choanocytes to microbial symbionts occurred over time, irrespective of microbial abundance, reflecting recycling of host waste products by the microbiome. Conclusions: Here, we provide empirical evidence indicating that the prokaryotic communities of a high and a low microbial abundance sponge obtain nutritional benefits from their host-associated lifestyle. The metabolic interaction between the highly efficient filter-feeding host and its microbial symbionts likely provides a competitive advantage to the sponge holobiont in the oligotrophic environments in which they thrive, by retaining and recycling limiting nutrients. Sponges present a unique model to link nutritional symbiotic interactions to holobiont function, and, via cascading effects, ecosystem functioning, in one of the earliest metazoan–microbe symbioses. 1KyCQaknthx73_ZUqh3faY Video abstract [ABSTRACT FROM AUTHOR]

CaMnt is much less important than NaMnt due to its limited commercial applications. The time-dependent property of NaMnt gel has been studied extensively as it is exploited in many applications such as drilling mud and viscosity-modifier applications. In contrast, the time-dependent property of CaMnt suspension and the factors affecting it are largely unknown. The speciation of Ca2+ ions is one such factor to be evaluated. In the current study, pH and solids concentration were examined and then used to validate a recent theory on clay gel time dependency. The results supported the theory that a strong electrostatic double layer (EDL) repulsion in the 3-D network is needed to reorganize the structure and drive it toward the state of minimum free energy. The 12 wt.% CaMnt (STx-1b) gel displayed time-dependent behavior at pH 5 but not at its natural pH of 8.4. At pH 5, the interlayer Ca2+ ions became fully hydrated and desorbed from the platelet surface. This enhanced the EDL repulsion between the platelets. At pH >6.5, the positively charged hydrolysis product Ca(OH)+ began to form and was adsorbed, partially neutralizing the permanent layer charge and weakening the repulsion. However, at 19.4 wt.% solids, the gel displayed pronounced time-dependent behavior despite the high natural pH of 8. The platelets were much closer together, allowing the EDL force to operate and effect structural reorganization. The microstructure of these CaMnt gels showed high platelet concentrations interacting to form a relatively open structure. The microstructure of a kaolin (KGa-2) suspension which showed no time-dependent behavior even after 1 day of ageing revealed the importance of particle morphology and layer charge on time dependency. Its 3-D structure was formed by relatively thick, layered platelets with a low layer charge. [ABSTRACT FROM AUTHOR]

Poly(2-hydroxyethyl methacrylate) hydrogels incorporating copper nanoparticles (Cu-PHEMA) were prepared by a two-step process. In the first step, porous PHEMA hydrogels containing aqueous Cu(II) were formed by photo-induced phase separation polymerization in aqueous formulations containing 2-hydroxyethyl methacrylate (HEMA), a photoinitator, a cross-linker, and CuSO4. In the second step, the Cu(II) ions were reduced to Cu(0) (Cu NPs) by hydrazine. The Cu-PHEMA hydrogels were red-brown due to the presence of the Cu NPs, and X-ray diffraction studies confirmed the NPs were crystalline Cu and not CuO. The porous PHEMA and Cu-PHEMA hydrogels exhibited the polymer droplet morphology characteristic of sponges formed by induced phase separation polymerization of HEMA in water. The Cu NPs were stable inside the hydrogel, and leaching of Cu NPs into water was very slow. Leached Cu NPs moderately inhibited the growth of Escherichia coli or Staphylococcus aureus bacteria in the surrounding medium. Incorporation of Cu NPs inside PHEMA hydrogels did not change compression strength of hydrogels. [ABSTRACT FROM AUTHOR]

The calcifuge habit of plants is commonly explained in terms of high soil pH and its effects on nutrient availability, particularly that of phosphorus (P). However, most Proteaceae that occur on nutrient‐impoverished soils in south‐western Australia are calcifuge, despite their ability to produce cluster‐roots, which effectively mobilize soil P and micronutrients. We hypothesize that the mechanism explaining the calcifuge habit in Proteaceae is their sensitivity to P and calcium (Ca), and that soil‐indifferent species are less sensitive to the interaction of these nutrients. In this study, we analyzed growth, gas‐exchange rate, and chlorophyll fluorescence of two soil‐indifferent and four calcifuge Hakea and Banksia (Proteaceae) species from south‐western Australia, across a range of P and Ca concentrations in hydroponic solution. We observed Ca‐enhanced P toxicity in all analyzed species, but to different extents depending on distribution type and genus. Increasing P supply enhanced plant growth, leaf biomass, and photosynthetic rates of soil‐indifferent species in a pattern largely independent of Ca supply. In contrast, positive physiological responses to increasing [P] in calcifuges were either absent or limited to low Ca supply, indicating that calcifuges were far more sensitive to Ca‐enhanced P toxicity. In calcifuge Hakeas, we attributed this to higher leaf [P], and in calcifuge Banksias to lower leaf zinc concentration. These differences help to explain these species' contrasting sensitivity to Ca‐enhanced P toxicity and account for the exclusion of most Proteaceae from calcareous habitats. We surmise that Ca‐enhanced P toxicity is a major factor explaining the calcifuge habit of Proteaceae, and, possibly, other P‐sensitive plants. [ABSTRACT FROM AUTHOR]

Lack of O2 and high concentrations of iron (Fe) and manganese (Mn) commonly occur in waterlogged soils. The development of a barrier to impede radial O2 loss (ROL) is a key trait improving internal O2 transport and waterlogging tolerance in plants. We evaluated the ability of the barrier to ROL to impede the entry of excess Fe into the roots of the waterlogging-tolerant grass Urochloa humidicola. Plants were grown in aerated or stagnant deoxygenated nutrient solution with 5 µM or 900 µM Fe. Quantitative X-ray microanalysis was used to determine cell-specific Fe concentrations at two positions behind the root apex in relation to ROL and the formation of apoplastic barriers. At a mature zone of the root, Fe was 'excluded' at the exodermis where a suberized lamella was evident, a feature also associated with a strong barrier to ROL. In contrast, the potassium (K) concentration was similar in all root cells, indicating that K uptake was not affected by apoplastic barriers. The hypothesis that the formation of a tight barrier to ROL impedes the apoplastic entry of toxic concentrations of Fe into the mature zones of roots was supported by the significantly higher accumulation of Fe on the outer side of the exodermis. [ABSTRACT FROM AUTHOR]

Very few of the >650 Proteaceae species in south-western Australia cope with the high calcium (Ca) levels in young, calcareous soils (soil indifferent); most are Ca sensitive and occur on nutrient-impoverished, acidic soils (calcifuge). We assessed possible control points for Ca transport across roots of two soil-indifferent (Hakea prostrata and Banksia prionotes) and two calcifuge (H. incrassata and B. menziesii) Proteaceae. Using quantitative X-ray microanalysis, we investigated cell-specific elemental Ca concentrations at two positions behind the apex in relation to development of apoplastic barriers in roots of plants grown in nutrient solution with low or high Ca supply. In H. prostrata , Ca accumulated in outer cortical cells at 20 mm behind the apex, but [Ca] was low in other cell types. In H. incrassata , [Ca] was low in all cells. Accumulation of Ca in roots of H. prostrata corresponded to development of apoplastic barriers in the endodermis. We found similar [Ca] profiles in roots and similar [Ca] in leaves of two contrasting Banksia species. Soil-indifferent Hakea and Banksia species show different strategies to inhabit calcareous soils: H. prostrata intercepts Ca in roots, reducing transport to shoots, whereas B. prionotes allocates Ca to specific leaf cells. [ABSTRACT FROM AUTHOR]

Sponge transcriptomes are important resources for studying the stress responses of these ecologically important filter feeders, the interactions between sponges and their symbionts, and the evolutionary history of metazoans. Here, we generated reference transcriptomes for two common and cosmopolitan Indo-Pacific sponge species: Carteriospongia foliascens and Cliona orientalis. We also created a reference transcriptome for the primary symbiont of C. orientalis—Gerakladium endoclionum. Assemblies for C. foliascens, C. orientalis, and G. endoclionum contained 67,304, 82,895, and 28,670 contigs, respectively. Contigs represented 15,248–37,344 isogroups (~ genes) per assembly, and N50s ranged from 1672–4355 bp. Sponge transcriptomes were high in completeness and quality, with an average of 93% of core EuKaryotic Orthologous Groups (KOGs) and 98% of single-copy metazoan core gene orthologs identified. The G. endoclionum assembly was partial with 56% of core KOGs and 32% of single-copy eukaryotic core gene orthologs identified. These reference transcriptomes provide a valuable resource for future research assessing sponge stress responses. [ABSTRACT FROM AUTHOR]

Summary: Endemism and rarity have long intrigued scientists. We focused on a rare endemic and critically‐endangered species in a global biodiversity hotspot, Grevillea thelemanniana (Proteaceae).We carried out plant and soil analyses of four Proteaceae, including G. thelemanniana, and combined these with glasshouse studies. The analyses related to hydrology and plant water relations as well as soil nutrient concentrations and plant nutrition, with an emphasis on sodium (Na) and calcium (Ca).The local hydrology and matching plant traits related to water relations partially accounted for the distribution of the four Proteaceae. What determined the rarity of G. thelemanniana, however, was its accumulation of Ca. Despite much higher total Ca concentrations in the leaves of the rare G. thelemanniana than in the common Proteaceae, very few Ca crystals were detected in epidermal or mesophyll cells. Instead of crystals, G. thelemanniana epidermal cell vacuoles contained exceptionally high concentrations of noncrystalline Ca. Calcium ameliorated the negative effects of Na on the very salt‐sensitive G. thelemanniana. Most importantly, G. thelemanniana required high concentrations of Ca to balance a massively accumulated feeding‐deterrent carboxylate, trans‐aconitate.This is the first example of a calcicole species accumulating and using Ca to balance accumulation of an antimetabolite. [ABSTRACT FROM AUTHOR]

Aims: Arbuscule-producing fine root endophytes (FRE) (previously incorrectly Glomus tenue) were recently placed within subphylum Mucoromycotina; the first report of arbuscules outside subphylum Glomeromycotina. Here, we aimed to estimate nutrient concentrations in plant and fungal structures of FRE and to test the utility of cryo-scanning electron microscopy (cryoSEM) for studying these fungi. Methods: We used replicated cryoSEM and X-ray microanalysis of heavily colonized roots of Trifolium subterraneum. Results: Intercellular hyphae and hyphae in developed arbuscules were consistently very thin; 1.35 ± 0.03 μm and 0.99 ± 0.03 μm in diameter, respectively (mean ± SE). Several intercellular hyphae were often adjacent to each other forming "hyphal ropes." Developed arbuscules showed higher phosphorus concentrations than senesced arbuscules and non-colonized structures. Senesced arbuscules showed greatly elevated concentrations of calcium and magnesium. Conclusion: While uniformly thin hyphae and hyphal ropes are distinct features of FRE, the morphology of fully developed arbuscules, elevated phosphorus in fungal structures, and accumulation of calcium with loss of structural integrity in senesced arbuscules are similar to glomeromycotinian fungi. Thus, we provide evidence that FRE may respond to similar host-plant signals or that the host plant may employ a similar mechanism of association with FRE and AMF. [ABSTRACT FROM AUTHOR]

Poly-(2-hydroxyethyl methacrylate) sponges doped with silver nanoparticles (Ag NP-PHEMA) were prepared by irradiation of aqueous solutions containing 2-hydroxyethyl methacrylate (HEMA) and silver nitrate in the presence of a cross-linking agent and a photoinitiator. The Ag NP-PHEMA sponges had polymer droplet morphology with interconnected pores, characteristic of PHEMA sponges prepared by polymerization-induced phase separation. The Ag NP-PHEMA sponges were yellow-brown, whereas PHEMA sponges were white. For formation of Ag NP-PHEMA, the size of the polymer droplets was larger and the rate of polymerization was increased compared to PHEMA sponges formed under similar conditions but in the absence of silver nitrate. Mechanical strength, as indicated by compression testing, was similar for Ag NP-PHEMA and PHEMA sponges. Nanoparticles slowly leached out of Ag NP-PHEMA sponges, exerting antibacterial activity against Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacteria. Notably, Ag NPs that leached were nontoxic to human corneal epithelial (HCE-T) cells. [ABSTRACT FROM AUTHOR]

Aims: High pH, and high bicarbonate (HCO3−) and calcium (Ca) availability characterise calcareous soils. High [Ca] only partially explains why some Lupinus species are calcifuge, so we explored high [HCO3−] and high pH. Methods: We grew six Lupinus genotypes in hydroponics with pH 5, 6.5 and 8a (adjusted by KOH), and 8b (adjusted by KHCO3). Leaf symptoms and areas, root appearance and biomass were recorded; whole leaf and root nutrient concentrations, and leaf cellular phosphorus (P), Ca and potassium (K) concentrations were determined using elemental X-ray microanalysis. Results: Chlorosis was observed in young leaves at high pH for L. angustifolius and L. cosentinii, and P deficiency at high pH for all genotypes. High pH decreased iron (Fe) and zinc (Zn) uptake in all genotypes. It also decreased lateral root growth, the uptake of P, K, Ca, and manganese (Mn) by all sensitive species; and translocation of P, Fe, Zn, Mn, and Ca to leaves in most sensitive species. However, leaf [Ca], leaf [K], [K] within each measured cell type, and translocation of K and Ca to leaves of L. pilosus and L. cosentinii at pH 8 were greater than at pH 5 and 6.5. Compared with pH 8a, all L. angustifolius genotypes translocated more P, Fe, Zn, Mn and K from roots to leaves at pH 8b. High pH did not affect the leaf cell types that accumulated P and Ca, but decreased the leaf cellular [P]. Conclusions: Lupinus angustifolius and L. cosentinii were sensitive to high [HCO3−] and/or high pH; L. pilosus was relatively tolerant. High pH decreased lateral root growth and nutrient uptake, inhibiting growth of sensitive species. High [HCO3−] diminished the negative effect of pH 8 on nutrient translocation to leaves in most L. angustifolius genotypes. This knowledge provides critical insights into the habits of Lupinus species to guide breeding of calcicole plants. [ABSTRACT FROM AUTHOR]

The rheological properties of sepiolite gels in relation to solution chemistry, fiber charge, and microstructure are poorly understood. The purpose of this study was to bring more clarity to this topic by quantifying the effects of solution pH, ionic strength, and adsorbed tetrasodium pyrophosphate (TSPP) additive on rheological properties. The electrical charge on sepiolite fibers was investigated to explain the fiber interaction configuration observed in the microstructure. Fiber interaction forces and dynamics explained the ageing behavior of the gel. Sepiolite gels of only a few percent solids displayed long-time ageing behavior, which was manifested by an increasing yield stress with wait time and continued for weeks. The gel microstructure showed randomly orientated rigid fibers with cross configuration attraction. Each fiber experiences both attractive (van der Waals and heterogeneous charge) and repulsive (electric double layer) forces, and initially a net force. The repulsive force causes these fibers to orientate or move continually to achieve a state of force equilibrium and this process takes a long time. The Leong model describes this ageing behavior. For good fiber separation, high intensity probe sonication of the suspension was required. The yield stress increased with sonication time, solids loading, and temperature. The yield stress was absent at pH > 11 and increased to a maximum value at pH < 8. This maximum was insensitive to pH between 4 to 8, and ionic strength up to 1 M KCl. TSPP reduced this maximum and shifted the zero yield stress region to a lower pH, ~7. The zero yield stress state corresponded to a zeta potential with a minimum magnitude of 30 mV. [ABSTRACT FROM AUTHOR]

In contrast to the situation in plants inhabiting most of the world's ecosystems, mycorrhizal fungi are usually absent from roots of the only two native vascular plant species of maritime Antarctica, Deschampsia antarctica and Colobanthus quitensis. Instead, a range of ascomycete fungi, termed dark septate endophytes (DSEs), frequently colonise the roots of these plant species. We demonstrate that colonisation of Antarctic vascular plants by DSEs facilitates not only the acquisition of organic nitrogen as early protein breakdown products, but also as non‐proteinaceous d‐amino acids and their short peptides, accumulated in slowly‐decomposing organic matter, such as moss peat. Our findings suggest that, in a warming maritime Antarctic, this symbiosis has a key role in accelerating the replacement of formerly dominant moss communities by vascular plants, and in increasing the rate at which ancient carbon stores laid down as moss peat over centuries or millennia are returned to the atmosphere as CO2. [ABSTRACT FROM AUTHOR]

Salinity tolerance is associated with Na 'exclusion' from, or 'tissue tolerance' in, leaves. We investigated whether two contrasting chickpea genotypes, salt-tolerant Genesis836 and salt-sensitive Rupali, differ in leaf tissue tolerance to NaCl. We used X-ray microanalysis to evaluate cellular Na, Cl, and K concentrations in various cell types within leaflets and also in secretory trichomes of the two chickpea genotypes in relation to photosynthesis in control and saline conditions. TEM was used to assess the effects of salinity on the ultrastructure of chloroplasts. Genesis836 maintained net photosynthetic rates (A) for the 21 d of salinity treatment (60 mM NaCl), whereas A in Rupali substantially decreased after 11 d. Leaflet tissue [Na] was low in Genesis836 but had increased markedly in Rupali. In Genesis836, Na was accumulated in epidermal cells but was low in mesophyll cells, whereas in Rupali cellular [Na] was high in both cell types. The excessive accumulation of Na in mesophyll cells of Rupali corresponded to structural damage to the chloroplasts. Maintenance of photosynthesis and thus salinity tolerance in Genesis836 was associated with an ability to 'exclude' Na from leaflets and in particular from the photosynthetically active mesophyll cells, and to compartmentalize Na in epidermal cells. [ABSTRACT FROM AUTHOR]

Over 650 Proteaceae occur in south-western Australia, contributing to the region's exceptionally high biodiversity. Most Proteaceae occur exclusively on severely nutrient-impoverished, acidic soils (calcifuge), whilst only few also occur on young, calcareous soils (soil-indifferent), higher in calcium (Ca) and phosphorus (P). The calcifuge habit of Proteaceae is explained by Ca-enhanced P toxicity, putatively linked to the leaf cell-specific allocation of Ca and P. Separation of these elements is essential to avoid the deleterious precipitation of Ca-phosphate. We used quantitative X-ray microanalysis to determine leaf cell-specific nutrient concentrations of two calcifuge and two soil-indifferent Proteaceae grown in hydroponics at a range of Ca and P concentrations. Calcium enhanced the preferential allocation of P to palisade mesophyll (PM) cells under high P conditions, without a significant change in whole leaf [P]. Calcifuges showed a greater PM [P] compared with soil-indifferent species, corresponding to their greater sensitivity. This study advances our mechanistic understanding of Ca-enhanced P toxicity, supporting the proposed model, and demonstrating its role in the calcifuge distribution of Proteaceae. This furthers our understanding of nutrient interactions at the cellular level and highlights its importance to plant functioning. [ABSTRACT FROM AUTHOR]

The Jurien Bay dune chronosequence in south‐western Australia's biodiversity hotspot comprises sites differing in nutrient availability, with phosphorus (P) availability declining strongly with increasing soil age. We have explored the exceptionally high photosynthetic P‐use efficiency (PPUE) of Proteaceae in this region, triggering the question what the PPUE of co‐occurring species in other families might be along the Jurien Bay chronosequence.We explored how traits associated with PPUE, photosynthetic nitrogen (N)‐use efficiency (PNUE) and leaf respiration might converge along the chronosequence, and whether Proteaceae and non‐Proteaceae species differ in leaf traits associated with nutrient use.Seven to 10 species were sampled at three sites differing in nutrient availability (ranging from N‐ to P‐limited). Measurements of leaf light‐saturated photosynthesis and dark respiration were integrated with measurements of total N and P concentration in both mature and senesced leaves, and leaf mass per unit area (LMA).Contrary to what is known for other chronosequences, rates of photosynthesis and respiration did not decrease with increasing soil age and LMA along the Jurien Bay chronosequence. However, they increased when expressed per unit leaf P. Both N and P were used much more efficiently for photosynthesis on nutrient‐poor sites, in both Proteaceae and non‐Proteaceae species. Proteaceae had the fastest rate of photosynthesis per unit leaf P, followed by species that preferentially allocate P to mesophyll cells, rather than epidermal cells.Synthesis. Our results show that with declining soil P availability, photosynthetic P‐use efficiency of all investigated species from different families increased. Plants growing on the oldest, most nutrient‐impoverished soils exhibited similar rates of CO2 exchange as plants growing on more nutrient‐rich younger soils, and extraordinarily high photosynthetic P‐use efficiency. This indicates convergence in leaf traits related to photosynthetic nutrient use on severely P‐impoverished sites. [ABSTRACT FROM AUTHOR]

Crops with improved uptake of fertilizer phosphorus (P) would reduce P losses and confer environmental benefits. We examined how P‐sufficient 6‐week‐old soil‐grown Trifolium subterraneum plants, and 2‐week‐old seedlings in solution culture, accumulated P in roots after inorganic P (Pi) addition. In contrast to our expectation that vacuoles would accumulate excess P, after 7 days, X‐ray microanalysis showed that vacuolar [P] remained low (<12 mmol kg−1). However, in the plants after P addition, some cortex cells contained globular structures extraordinarily rich in P (often >3,000 mmol kg−1), potassium, magnesium, and sodium. Similar structures were evident in seedlings, both before and after P addition, with their [P] increasing threefold after P addition. Nuclear magnetic resonance (NMR) spectroscopy showed seedling roots accumulated Pi following P addition, and transmission electron microscopy (TEM) revealed large plastids. For seedlings, we demonstrated that roots differentially expressed genes after P addition using RNAseq mapped to the T. subterraneum reference genome assembly and transcriptome profiles. Among the most up‐regulated genes after 4 hr was TSub_g9430.t1, which is similar to plastid envelope Pi transporters (PHT4;1, PHT4;4): expression of vacuolar Pi‐transporter homologs did not change. We suggest that subcellular P accumulation in globular structures, which may include plastids, aids cytosolic Pi homeostasis under high‐P availability. We examined clover roots after inorganic phosphorus (Pi) addition. Although vacuolar P concentration was unchanged, unexpectedly, P accumulated in intracellular globular structures and genes similar to plastid envelope Pi transporters were up‐regulated. The structures likely aid cytosolic Pi homeostasis. [ABSTRACT FROM AUTHOR]

Summary: Many Proteaceae are highly phosphorus (P)‐sensitive and occur exclusively on old nutrient‐impoverished acidic soils (calcifuge), whilst a few also occur on young calcareous soils (soil‐indifferent) that are higher in available calcium (Ca) and P. Calcium increases the severity of P‐toxicity symptoms, but its underlying mechanisms are unknown. We propose that Ca‐enhanced P toxicity explains the calcifuge habit of most Proteaceae.Four calcifuge and four soil‐indifferent Proteaceae from South‐Western Australia were grown in hydroponics, at a range of P and Ca concentrations.Calcium increased the severity of P‐toxicity symptoms in all species. Calcifuge Proteaceae were more sensitive to Ca‐enhanced P toxicity than soil‐indifferent ones. Calcifuges shared these traits: low leaf zinc concentration ([Zn]), low Zn allocation to leaves, low leaf [Zn]:[P], low root : shoot ratio, and high seed P content, compared with soil‐indifferent species.This is the first demonstration of Ca‐enhanced P toxicity across multiple species. Calcium‐enhanced P toxicity provides an explanation for the calcifuge habit of most Proteaceae and is critical for the management of this iconic Australian family. This study represents a major advance towards an understanding of the physiological mechanisms of P toxicity and its role in the distribution of Proteaceae. [ABSTRACT FROM AUTHOR]

Aims: Previous studies have shown that pH, rather than calcium (Ca), is the main reason why some Lupinus species are sensitive to nutrient solutions mimicking calcareous soils; however, a hydroponic system is quite different from soil systems, and plants may respond differently to these two growing conditions. Thus, studies with Lupinus species grown in calcareous soils are needed.Methods: Two calcicole and two calcifuge species were grown in river sand with different Ca forms and amounts, pH levels, and [bicarbonate (HCO3−)] (HCO3− concentration, which is produced by calcium carbonate (CaCO3)). Leaf symptoms, leaf area, gas exchange, biomass, and root morphology were recorded; whole leaf and root nutrient concentrations were analysed.Results: We observed leaf chlorosis of the youngest leaves under high pH (adjusted by KOH) and high pH + high Ca (representing high [HCO3−], high pH and high Ca) treatments for all Lupinus species. However, after 2 weeks, leaf chlorosis of all Lupinus species under high pH started to disappear, with calcicole species fully, and calcifuge species only partly recovering. Leaf chlorosis symptoms of calcicole species under high pH + high Ca partly disappeared as well, while those of calcifuge species did not disappear at all.Conclusions: High pH (resulting from either KOH or HCO3−) inhibited root growth, and subsequently uptake of some nutrients and shoot growth of Lupinus species. However, the strong buffering capacity of HCO3− is the key factor determining if Lupinus species can survive in calcareous soils. Among all studied Lupinus species, L. pilosus was the most tolerant to high [HCO3−] and/or high pH, followed by L. cosentinii and L. angustifolius, while L. hispanicus was the most sensitive. [ABSTRACT FROM AUTHOR]

Microorganisms drive most of the major biogeochemical cycles in the ocean, but the rates at which individual species assimilate and transform key elements is generally poorly quantified. One of these important elements is nitrogen, with its availability limiting primary production across a large proportion of the ocean. Nitrogen uptake by marine microbes is typically quantified using bulk-scale approaches, such as Elemental Analyzer-Isotope Ratio Mass Spectrometry (EA-IRMS), which averages uptake over entire communities, masking microbial heterogeneity. However, more recent techniques, such as secondary ion mass spectrometry (SIMS), allow for elucidation of assimilation rates at the scale at which they occur: the single-cell level. Here, we combine and compare the application of bulk (EA-IRMS) and single-cell approaches (NanoSIMS and Time-of-Flight-SIMS) for quantifying the assimilation of inorganic nitrogen by the ubiquitous marine primary producer Synechococcus. We aimed to contrast the advantages and disadvantages of these techniques and showcase their complementarity. Our results show that the average assimilation of 15N by Synechococcus differed based on the technique used: values derived from EA-IRMS were consistently higher than those derived from SIMS, likely due to a combination of previously reported systematic depletion as well as differences in sample preparation. However, single-cell approaches offered additional layers of information, whereby NanoSIMS allowed for the quantification of the metabolic heterogeneity among individual cells and ToF-SIMS enabled identification of nitrogen assimilation into peptides. We suggest that this coupling of stable isotope-based approaches has great potential to elucidate the metabolic capacity and heterogeneity of microbial cells in natural environments. [ABSTRACT FROM AUTHOR]

Natural (N-) and synthetic (S-) hectorite suspensions were found to display significant time-dependent rheology or ageing behaviour and shear thinning flow behaviour. The ageing behaviour was characterised by an increasing yield stress with rest time. The yield stress continued to increase even after a week of rest, a reflection of a long process. An open sponge-like cellular microstructure formed by platelet particles interacting attractively in the overlapping edge-face configuration was captured by cryo-SEM of gel samples prepared at high pressure (∼2000 bar) and subjected to rapid cryo-freezing, for both N- and S-hectorite gels. Even nano-discotic S-hectorite particles formed platelet particles hundreds of nanometres in length in the overlapping coin configuration. This structure, displaying a cell size ranging from tens to several hundred nanometres, is formed by strong attractive and repulsive forces. The platelets showed deformations such as bending and curling of the edges in response to these forces. The S-hectorite platelets are smaller and more rigid. During ageing the particles in the structure experience a net force. These particles will move in response causing force imbalance to be experienced by neighbouring particles and they will move in response. This action and reaction percolate through the network structure causing a high concentration of particles to respond. As a consequence the ageing process takes a long time to reach equilibrium. Various ageing models were used to fit the ageing data. The N-hectorite gels displayed a maximum yield stress at pH ∼ 8 and a particle zeta potential of −35 mV. This suggests the presence of critical positive and negative charge density is needed to form a structure with maximum strength. The zeta potential is negative and quite insensitive to pH from pH 4 to 12. [ABSTRACT FROM AUTHOR]

Phosphorus (P)‐deficiency symptoms are known for Lupinus species grown in calcareous soil, but we do not know if this is due to high calcium (Ca) availability or low P availability in the soil. To address this problem, we explored both the effects of Ca and its interaction with P on nutrient status and growth of three Lupinus species. Two Ca‐sensitive genotypes (Lupinus angustifolius L. P26723 and Lupinus cosentinii Guss. P27225) and two Ca‐tolerant genotypes (L. angustifolius L. cv Mandelup and Lupinus pilosus Murr. P27440) were grown hydroponically at two P (0.1 and 10 μM) and three Ca (0.1, 0.6 and 6 mM) levels. Leaf symptoms and biomass were recorded, whole leaf and root nutrient concentrations were analysed, and leaf cellular P and Ca concentrations were determined. Phosphorus‐deficiency symptoms were only observed in the Ca‐sensitive genotypes. Among all the genotypes in this study, the Ca‐tolerant L. pilosus showed an ability to maintain stable leaf Ca and P concentrations whereas the Ca‐tolerant L. angustifolius cv Mandelup did not maintain a stable whole leaf Ca concentration, but maintained a low cytosolic Ca2+ concentration through effective Ca compartmentation. However, the two Ca‐sensitive genotypes, L. angustifolius P26723 and L. cosentinii, did not exhibit an ability to maintain a stable whole leaf Ca concentration or effectively compartmentalize Ca. Therefore, having the capacity to maintain a stable whole leaf Ca concentration or effectively compartmentalizing Ca in leaves are likely critical for Lupinus species to be Ca tolerant. [ABSTRACT FROM AUTHOR]

Abstract: To study mechanism underpinning the calcifuge habit of some Lupinus species, especially under low‐phosphorus (P) conditions, Lupinus species that were likely to respond differently to calcium (Ca) availability were assembled, and the sensitivity to Ca under a low‐P supply was assessed. Seven Lupinus species (9 genotypes, L. albus L. cv Kiev, L. albus L. P26766, L. angustifolius L. cv Mandelup, L. angustifolius L. P26723, L. luteus L. cv Pootalong, L. hispanicus ssp. bicolor Boiss. and Reut. P22999, L. pilosus Murr. P27440, L. cosentinii Guss. P27225, and L. atlanticus Gladst. P27219) were grown hydroponically at 10 or 6000 μM Ca. Leaf symptoms, gas exchange and biomass were recorded; leaf and root nutrient concentrations were analysed, and the leaf cell types in which Ca and P accumulated were determined using elemental X‐ray microanalyses. Calcium toxicity was demonstrated for L. angustifolius P26723, L. hispanicus ssp. bicolor. P22999, and L. cosentinii P27225, whereas the other species were tolerant of a high Ca supply under low‐P conditions. In addition, genotypic differences in Ca toxicity were found within L. angustifolius. Most Ca accumulated in the mesophyll cells in all species, whereas most P was located in epidermal cells. [ABSTRACT FROM AUTHOR]

Summary: Plants allocate nutrients to specific leaf cell types, with commelinoid monocots preferentially allocating phosphorus (P) to the mesophyll and calcium (Ca) to the epidermis, whereas the opposite is thought to occur in eudicots. However, Proteaceae from severely P‐impoverished habitats present the same P‐allocation pattern as monocots. This raises the question of whether preferential P allocation to mesophyll cells is a phylogenetically conserved trait, exclusive to commelinoid monocots and a few Proteaceae, or a trait that has evolved multiple times to allow plants to cope with very low soil P availability. We analysed the P‐allocation patterns of 16 species from 10 genera, eight families and six orders within three major clades of eudicots across different P‐impoverished environments in Australia and Brazil, using elemental X‐ray mapping to quantitatively determine leaf cell‐specific nutrient concentrations. Many of the analysed species showed P‐allocation patterns that differed substantially from that expected for eudicots. Instead, P‐allocation patterns were strongly associated with the P availability in the natural habitat of the species, suggesting a convergent evolution of P‐allocation patterns at the cellular level, with P limitation as selective pressure and without a consistent P‐allocation pattern within eudicots. Here, we show that most eudicots from severely P‐impoverished environments preferentially allocated P to their mesophyll. We surmise that this preferential P allocation to photosynthetically active cells might contribute to the very high photosynthetic P‐use efficiency of species adapted to P‐impoverished habitats. [ABSTRACT FROM AUTHOR]

Abstract: Plants allocate nutrients to specific leaf cell types; eudicots are thought to predominantly allocate phosphorus (P) to epidermal/bundle sheath cells. However, three Proteaceae species have been shown to preferentially allocate P to mesophyll cells instead. These Proteaceae species are highly adapted to P‐impoverished habitats, with exceptionally high photosynthetic P‐use efficiencies (PPUE). We hypothesized that preferential allocation of P to photosynthetic mesophyll cells is an important trait in species adapted to extremely P‐impoverished habitats, contributing to their high PPUE. We used elemental X‐ray mapping to determine leaf cell‐specific nutrient concentrations for 12 Proteaceae species, from habitats of strongly contrasting soil P concentrations, in Australia, Brazil, and Chile. We found that only species from extremely P‐impoverished habitats preferentially allocated P to photosynthetic mesophyll cells, suggesting it has evolved as an adaptation to their extremely P‐impoverished habitat and that it is not a family‐wide trait. Our results highlight the possible role of soil P in driving the evolution of ecologically relevant nutrient allocation patterns and that these patterns cannot be generalized across families. Furthermore, preferential allocation of P to photosynthetic cells may provide new and exciting strategies to improve PPUE in crop species. [ABSTRACT FROM AUTHOR]

Terrestrial runoff, resuspension events and dredging can affect filter-feeding sponges by elevating the concentration of suspended sediments, reducing light intensity, and smothering sponges with sediments. To investigate how sponges respond to pressures associated with increased sediment loads, the abundant and widely distributed Indo-Pacific species Ianthella basta was exposed to elevated suspended sediment concentrations, sediment deposition, and light attenuation for 48 h (acute exposure) and 4 weeks (chronic exposure). In order to visualise the response mechanisms, sponge tissue was examined by 3D X-ray microscopy and scanning electron microscopy (SEM). Acute exposures resulted in sediment rapidly accumulating in the aquiferous system of I. basta, although this sediment was fully removed within three days. Sediment removal took longer (>2 weeks) following chronic exposures, and I. basta also exhibited tissue regression and a smaller aquiferous system. The application of advanced imaging approaches revealed that I. basta employs a multilevel system for sediment rejection and elimination, containing both active and passive components. Sponges responded to sediment stress through (i) mucus production, (ii) exclusion of particles by incurrent pores, (iii) closure of oscula and pumping cessation, (iv) expulsion of particles from the aquiferous system, and (v) tissue regression to reduce the volume of the aquiferous system, thereby entering a dormant state. These mechanisms would result in tolerance and resilience to exposure to variable and high sediment loads associated with both anthropogenic impacts like dredging programs and natural pressures like flood events. [ABSTRACT FROM AUTHOR]

Reef corals are under threat from bleaching and disease outbreaks that target both the host animal and the algal symbionts within the coral holobiont. A viral origin for coral bleaching has been hypothesized, but direct evidence has remained elusive. Using a multifaceted approach incorporating flow cytometry, transmission electron microscopy, DNA and RNA virome sequencing, we show that type C1 Symbiodinium cultures host a nucleocytoplasmic large double-stranded DNA virus (NCLDV) related to Phycodnaviridae and Mimiviridae, a novel filamentous virus of unknown phylogenetic affiliation, and a single-stranded RNA virus related to retroviruses. We discuss implications of these findings for laboratory-based experiments using Symbiodinium cultures. [ABSTRACT FROM AUTHOR]

Background: Molecular and structural information on avian Trypanosoma spp. throughout Australia is limited despite their intrinsic value in understanding trypanosomatid evolution, diversity, and structural biology. In Western Australia tissue samples (n = 429) extracted from 93 birds in 25 bird species were screened using generic PCR primers to investigate the diversity of Trypanosoma spp. To investigate avian trypanosome structural biology the first 3-dimensional ultrastructural models of a Trypanosoma spp. (Trypanosoma sp. AAT) isolated from a bird (currawong, Strepera spp.) were generated using focussed ion beam milling combined with scanning electron microscopy (FIB-SEM). Results: Here, we confirm four intercontinental species of avian trypanosomes in native Australian birds, and identify a new avian Trypanosoma. Trypanosome infection was identified in 18 birds from 13 different bird species (19%). A single new genotype was isolated and found to be closely related to T. culicavium (Trypanosoma sp. CC2016 B002). Other Trypanosoma spp. identified include T. avium, T. culicavium, T. thomasbancrofti, Trypanosoma sp. TL.AQ.22, Trypanosoma sp. AAT, and an uncharacterised Trypanosoma sp. (group C-III sensu Zidková et al. (Infect Genet Evol 12:102-112, 2012)), all previously identified in Australia or other continents. Serially-sectioning Trypanosoma sp. AAT epimastigotes using FIB-SEM revealed the disc-shaped kinetoplast pocket attached perpendicular to the branching mitochondrion. Additionally, the universal minicircle sequence within the kinetoplast DNA and the associated binding protein were determined in Trypanosoma sp. AAT. Conclusions: These results indicate that bird trypanosomes are relatively conserved across continents, while being locally diverse, which supports the hypothesis that bird trypanosomes exist as fewer species than described in the literature. Evidence exists that avian Trypanosoma spp. are infecting mammals and could be transmitted by haemadipsid leeches. Trypanosoma sp. AAT is most likely a separate species currently found only in Australia and the first 3-dimentional ultrastructural analysis of an avian trypanosome provides interesting information on their morphology and organelle arrangement. [ABSTRACT FROM AUTHOR]

Background: Taxonomic identification of ticks obtained during a longitudinal survey of the critically endangered marsupial, Bettongia penicillata Gray, 1837 (woylie, brush-tailed bettong) revealed a new species of Ixodes Latrielle, 1795. Here we provide morphological data for the female and nymphal life stages of this novel species (Ixodes woyliei n. sp.), in combination with molecular characterisation using the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). In addition, molecular characterisation was conducted on several described Ixodes species and used to provide phylogenetic context. Results: Ixodes spp. ticks were collected from the two remaining indigenous B. penicillata populations in southwestern Australia. Of 624 individual B. penicillata sampled, 290 (47%) were host to ticks of the genus Ixodes; specifically I. woyliei n. sp., I. australiensis Neumann, 1904, I. myrmecobii Roberts, 1962, I. tasmani Neumann, 1899 and I. fecialis Warburton & Nuttall, 1909. Of these, 123 (42%) were host to the newly described I. woyliei n. sp. In addition, 268 individuals from sympatric marsupial species (166 Trichosurus vulpecula hypoleucus Wagner, 1855 (brushtail possum), 89 Dasyurus geoffroii Gould, 1841 (Western quoll) and 13 Isoodon obesulus fusciventer Gray, 1841 (southern brown bandicoot)) were sampled for ectoparasites and of these, I. woyliei n. sp. was only found on two I. o. fusciventer. Conclusions: Morphological and molecular data have confirmed the first new Australian Ixodes tick species described in over 50 years, Ixodes woyliei n. sp. Based on the long-term data collected, it appears this tick has a strong predilection for B. penicillata, with 42% of Ixodes infections on this host identified as I. woyliei n. sp. The implications for this hostparasite relationship are unclear but there may be potential for a future co-extinction event. In addition, new molecular data have been generated for collected specimens of I. australiensis, I. tasmani and museum specimens of I. victoriensis Nuttall, 1916, which for the first time provides molecular support for the subgenus Endopalpiger Schulze, 1935 as initially defined. These genetic data provide essential information for future studies relying on genotyping for species identification or for those tackling the phylogenetic relationships of Australian Ixodes species. [ABSTRACT FROM AUTHOR]

Suspended sediments produced from dredging activities, or added to the sediment budget via river runoff, are a concern for marine resource managers. Understanding the impact of suspended sediments on critical life history stages of keystone species like corals is fundamental to effective management of coastlines and reefs. Coral embryos (Acropora tenuis and A. millepora) and larvae (A. tenuis, A. millepora and Pocillopora acuta) were subjected to a range of suspended sediment concentrations of different sediment types (siliciclastic and carbonate) to assess concentration-response relationships on ecologically relevant endpoints, including survivorship and ability to metamorphose. Embryos were subjected to short (12 h) suspended sediment exposures from ages of 3–12 hours old or a long (30 h) exposure at 6 hours old. Neither the survivorship nor metamorphosis function of embryos were significantly affected by realistic sediment exposures to ~1000 mg L-1. However, some embryos exhibited a previously undescribed response to dynamically suspended sediments, which saw 10% of the embryos form negatively buoyant cocoons at siliciclastic suspended sediment concentrations ≥35 mg L-1. Scanning electron and optical microscopy confirmed the presence of a coating on these embryos, possibly mucus with incorporated sediment particles. Cocoon formation was common in embryos but not in larvae, and occurred more often after exposure to siliciclastic rather than carbonate sediments. Once transferred into sediment-free seawater, functional ~36-h-old embryos began emerging from the cocoons, coinciding with cilia development. Ciliated (> 36-h-old) larvae exposed to suspended sediments for 60 h were also observed to secrete mucus and were similarly unaffected by suspended sediment concentrations to ~800 mg L-1. This study provides evidence that mucous secretion and cilia beating effectively protect coral embryos and larvae from suspended sediment and that these mechanisms may enhance their chances of successful recruitment. [ABSTRACT FROM AUTHOR]

Background and Aims Sclerotinia stem rot (SSR, Sclerotinia sclerotiorum) is a damaging disease of oilseed brassicas world-wide. Host resistance is urgently needed to achieve control, yet the factors that contribute to stem resistance are not well understood. This study investigated the mechanisms of resistance to SSR. Methods Stems of 5-week-old Brassica carinata, B. juncea and B. napus of known resistance were infected via filter paper discs impregnated with S. sclerotiorum mycelium under controlled conditions. Transverse sections of the stem and portions of the stem surface were examined using optical and scanning electron microscopy. The association of anatomical features with the severity of disease (measured by mean lesion length) was determined. Key Results Several distinct resistance mechanisms were recorded for the first time in these Brassica-pathogen interactions, including hypersensitive reactions and lignification within the stem cortex, endodermis and in tissues surrounding the lesions. Genotypes showing a strong lignification response 72 h post-infection (hpi) tended to have smaller lesions. Extensive vascular invasion by S. sclerotiorum was observed only in susceptible genotypes, especially in the vascular fibres and xylem. Mean lesion length was negatively correlated with the number of cell layers in the cortex, suggesting progress of S. sclerotiorum is impeded by more cell layers. Hyphae in the centre of lesions became highly vacuolate 72 hpi, reflecting an ageing process in S. sclerotiorum hyphal networks that was independent of host resistance. The infection process of S. sclerotiorum was analogous in B. carinata and B. napus. Infection cushions of the highly virulent isolate of S. sclerotiorum MBRS-1 were grouped together in dense parallel bundles, while hyphae in the infection cushions of a less aggressive isolate WW-3 were more diffuse, and this was unaffected by host genotype. Conclusions A variety of mechanisms contribute to host resistance against S. sclerotiorum across the three Brassica species. These complex interactions between pathogen and host help to explain variable expressions of resistance often observed in the field. [ABSTRACT FROM AUTHOR]

Oxygen deficiency associated with soil waterlogging adversely impacts root respiration and nutrient acquisition. We investigated the effects of O2 deficiency and salinity (100 mM NaCl) on radial O2 concentrations and cell-specific ion distributions in adventitious roots of barley ( Hordeum vulgare)., Microelectrode profiling measured O2 concentrations across roots in aerated, aerated saline, stagnant or stagnant saline media. X-ray microanalysis at two positions behind the apex determined the cell-specific elemental concentrations of potassium (K), sodium (Na) and chloride (Cl) across roots., Severe O2 deficiency occurred in the stele and apical regions of roots in stagnant solutions. O2 deficiency in the stele reduced the concentrations of K, Na and Cl in the pericycle and xylem parenchyma cells at the subapical region. Near the root apex, Na declined across the cortex in roots from the aerated saline solution but was relatively high in all cell types in roots from the stagnant saline solution., Oxygen deficiency has a substantial impact on cellular ion concentrations in roots. Both pericycle and xylem parenchyma cells are involved in energy-dependent K loading into the xylem and in controlling radial Na and Cl transport. At root tips, accumulation of Na in the outer cell layers likely contributed to reduction of Na in inner cells of the tips. [ABSTRACT FROM AUTHOR]