Evolution of the CD163 family and its relationship to the bovine gamma delta T cell co-receptor WC1

Background  

The scavenger receptor cysteine rich (SRCR) domain is an ancient and conserved protein domain. CD163 and WC1 molecules are classed together as group B SRCR superfamily members, along with Spα, CD5 and CD6, all of which are expressed by immune system cells. There are three known types of CD163 molecules in mammals, CD163A (M130, coded for by CD163), CD163b (M160, coded for by CD163L1) and CD163c-α (CD163L1 or SCART), while their nearest relative, WC1, is encoded by a multigene family so far identified in the artiodactyl species of cattle, sheep, and pigs.     

Plasma membrane Ca²+ transporters mediate virus-induced acquired resistance to oxidative stress

This paper reports the phenomenon of acquired cross‐tolerance to oxidative stress in plants and investigates the activity of specific Ca²⁺ transport systems mediating this phenomenon. Nicotiana benthamiana plants were infected with Potato virus X (PVX) and exposed to oxidative [either ultraviolet (UV‐C) or H₂O₂] stress. Plant adaptive responses were assessed by the combined application of a range of electrophysiological (non‐invasive microelectrode ion flux measurements), biochemical (Ca²⁺‐ and H⁺‐ATPase activity), imaging (fluorescence lifetime imaging measurements of changes in intracellular Ca²⁺ concentrations), pharmacological and cytological transmission electrone microscopy techniques. Virus‐infected plants had a better ability to control UV‐induced elevations in cytosolic‐free Ca²⁺ and prevent structural and functional damage of chloroplasts. Taken together, our results suggest a high degree of crosstalk between UV and pathogen‐induced oxidative stresses, and highlight the crucial role of Ca²⁺ efflux systems in acquired resistance to oxidative stress in plants.     

Genetic behaviour of physiological traits conferring cytosolic K+/Na+ homeostasis in wheat

A plant's ability to maintain an optimal cytosolic K(+)/Na(+) ratio has long been cited as a key feature of salinity tolerance. As traditional whole-leaf nutrient analysis does not account for tissue and organelle-specific ion sequestration, the predictive value of this index at the whole-plant level is not always satisfactory. Consequently, suitable in situ methods for functionally assessing the activity of the key membrane transporters contributing to this trait at the cellular level need to be developed. The aim of this work was to investigate the extent to which plasma membrane transporter-mediated Na(+) exclusion and KOR-mediated K(+) retention traits, measured with the microelectrode ion flux measuring (MIFE) technique, are inheritable in wheat, and whether the MIFE technique has the potential to be used in combination with molecular markers to determine QTLs for these transporter proteins. Experiments involved two bread (Triticum aestivum) and two durum (Triticum turgidum) wheat lines contrasting in their salinity tolerance. Net Na(+), K(+) and H(+) fluxes were measured from 6-day-old roots of parental lines and their F(1) hybrids upon addition and removal of NaCl. These results were complemented by assessment of whole-plant physiological and agronomic characteristics. We show evidence for a strong heritability of plasma membrane transporter-mediated Na(+) exclusion and K(+) retention traits in wheat at the cellular level. This opens the prospect of using the MIFE technique to map the position of these transporters on particular loci of wheat chromosomes. The next obvious step would be to pyramid these traits in one ideotype with superior salinity tolerance.     

A root's ability to retain K+ correlates with salt tolerance in wheat

Most work on wheat breeding for salt tolerance has focused mainly on excluding Na(+) from uptake and transport to the shoot. However, some recent findings have reported no apparent correlation between leaf Na(+) content and wheat salt tolerance. Thus, it appears that excluding Na(+) by itself is not always sufficient to increase plant salt tolerance and other physiological traits should also be considered. In this work, it was investigated whether a root's ability to retain K(+) may be such a trait, and whether our previous findings for barley can be extrapolated to species following a 'salt exclusion' strategy. NaCl-induced kinetics of K(+) flux from roots of two bread and two durum wheat genotypes, contrasting in their salt tolerance, were measured under laboratory conditions using non-invasive ion flux measuring (the MIFE) technique. These measurements were compared with whole-plant physiological characteristics and yield responses from plants grown under greenhouse conditions. The results show that K(+) flux from the root surface of 6-d-old wheat seedlings in response to salt treatment was highly correlated with major plant physiological characteristics and yield of greenhouse-grown plants. This emphasizes the critical role of K(+) homeostasis in plant salt tolerance and suggests that using NaCl-induced K(+) flux measurements as a physiological 'marker' for salt tolerance may benefit wheat-breeding programmes.     

Detection of protease-resistant cervid prion protein in water from a CWD-endemic area

Chronic wasting disease (CWD) is the only known transmissible spongiform encephalopathy affecting free-ranging wildlife. Although the exact mode of natural transmission remains unknown, substantial evidence suggests that prions can persist in the environment, implicating components thereof as potential prion reservoirs and transmission vehicles.^1–4 CWD-positive animals may contribute to environmental prion load via decomposing carcasses and biological materials including saliva, blood, urine and feces.^5–7 Sensitivity limitations of conventional assays hamper evaluation of environmental prion loads in soil and water. Here we show the ability of serial protein misfolding cyclic amplification (sPMCA) to amplify a 1.3 × 10⁻⁷ dilution of CWD-infected brain homogenate spiked into water samples, equivalent to approximately 5 × 10⁷ protease resistant cervid prion protein (PrP^CWD) monomers. We also detected PrP^CWD in one of two environmental water samples from a CWD endemic area collected at a time of increased water runoff from melting winter snow pack, as well as in water samples obtained concurrently from the flocculation stage of water processing by the municipal water treatment facility. Bioassays indicated that the PrP^CWD detected was below infectious levels. These data demonstrate detection of very low levels of PrP^CWD in the environment by sPMCA and suggest persistence and accumulation of prions in the environment that may promote CWD transmission.Key words: prions, chronic wasting disease, water, environment, serial protein misfolding cyclic amplification     

Freshwater crayfish invasions in South Africa: past,present and potential future

Freshwater crayfish invasions have been studied around the world, but less so in Africa, a continent devoid of native freshwater crayfish. The present study reviews historical and current information on alien freshwater crayfish species introduced into South Africa and aims to indicate which areas are at risk from invasion. As is the case elsewhere, South Africans have shown a keen interest in both farming and keeping freshwater crayfish as pets, which has resulted in Cherax cainii, Cherax destructor, Cherax quadricarinatus and Procambarus clarkii being introduced to the country. There is evidence of successful establishment in the wild for C. quadricarinatus and P. clarkii in different parts of the country. Species distribution models suggest that the eastern part of the country and parts of the Eastern and Western Cape are at higher risk of invasion. At present, illegal translocations represent the most likely pathway of crayfish spread in South Africa. A continued risk of invasion by freshwater crayfish species in South Africa is highlighted, which reinforces the need for more research, as well as for strong mitigation measures, such as stronger policing of existing regulations, management or eradication where feasible and public education.     

Potassium transport and plant salt tolerance

Salinity is a major abiotic stress affecting approximately 7% of the world's total land area resulting in billion dollar losses in crop production around the globe. Recent progress in molecular genetics and plant electrophysiology suggests that the ability of a plant to maintain a high cytosolic K⁺/Na⁺ ratio appears to be critical to plant salt tolerance. So far, the major efforts of plant breeders have been aimed at improving this ratio by minimizing Na⁺ uptake and transport to shoot. In this paper, we discuss an alternative approach, reviewing the molecular and ionic mechanisms contributing to potassium homeostasis in salinized plant tissues and discussing prospects for breeding for salt tolerance by targeting this trait. Major K⁺ transporters and their functional expression under saline conditions are reviewed and the multiple modes of their control are evaluated, including ameliorative effects of compatible solutes, polyamines and supplemental calcium. Subsequently, the genetic aspects of inheritance of K⁺ transport 'markers' are discussed in the general context of salt tolerance as a polygenic trait. The molecular identity of 'salt tolerance' genes is analysed, and prospects for future research and breeding are examined.     

Nanolitre-scale assays to determine the activities of enzymes in individual plant cells

There are a variety of methods for characterising gene expression at the level of individual cells and for demonstrating that the cells also contain the encoded proteins. However, measuring the activity of enzymes at the resolution of single cells in complex tissues, such as leaves, is problematic. We have addressed this by using single-cell sampling to extract 10-100 pl droplets of sap from individual plant cells and then measuring enzyme activities in these droplets with nanolitre-scale fluorescence-based assays. We have optimised these assays and used them to measure and characterise the activities of acid phosphatase, cysteine protease and nitrate reductase in sap samples from epidermal and mesophyll cells of barley (Hordeum vulgare L.) and Arabidopsis thaliana leaves exposed to different developmental and environmental conditions. During leaf senescence in barley, we found that the dynamics with which acid phosphatase and protease activities changed were different in each cell type and did not mirror the changes occurring at the whole-leaf level. Increases in nitrate reductase activities after exposure of barley plants to nitrate were large in mesophyll cells but small in epidermal cells. The technique was applied successfully to Arabidopsis and, as in barley, revealed cell-specific differences in the activities of both acid phosphatase and nitrate reductase. The assays add to the spectrum of techniques available for characterising cells within complex plant tissues, thus extending the opportunity to relate gene expression to biochemical activities at the single-cell level.     

Potassium activities in cell compartments of salt-grown barley leaves

Triple-barrelled microelectrodes measuring K(+) activity (a(K)), pH and membrane potential were used to make quantitative measurements of vacuolar and cytosolic a(K) in epidermal and mesophyll cells of barley plants grown in nutrient solution with 0 or 200 mM added NaCl. Measurements of a(K) were assigned to the cytosol or vacuole based on the pH measured. In epidermal cells, the salt treatment decreased a(K) in the vacuole from 224 to 47 mM and in the cytosol from 68 to 15 mM. In contrast, the equivalent changes in the mesophyll were from 235 to 150 mM (vacuole) and 79 to 64 mM (cytosol). Thus mechanisms exist to ameliorate the effects of salt on a(K) in compartments of mesophyll cells, presumably to minimize any deleterious consequences for photosynthesis. Thermodynamic calculations showed that K(+) is actively transported into the vacuole of both epidermal and mesophyll cells of salinized and non- salinized plants. Comparison of the values of a(K) in K(+)-replete, non-salinized leaf cells with those previously measured in root cells of plants grown under comparable conditions indicates that cytosolic a(K) is similar in cells of both organs, but vacuolar a(K) in leaf cells is approximately twice that in roots. This suggests differences in the regulation of vacuolar a(K), but not cytosolic a(K), in leaf and root cells.