Red elemental selenium (Se0) improves the immunoactivities of EPC cells,crucian carp and zebrafish against spring viraemia of carp virus

Selenium, as an essential trace element, interferes through selenoproteins in many physiological processes of plants and mammals. Its antiviral activity has recently attracted much attention because selenium improves the antiviral capacity of animal cells against a few viruses relevant to human diseases. In this study, the red elemental selenium was purified from the fermentative culture of Herbaspirillum camelliae WT00C and then used to culture epithelioma papulosum cyprinid (EPC) cells or feed crucian carp and zebrafish. Finally, its antiviral effects were investigated at the cell level and living fishes after spring viraemia of carp virus infection. At the cell level, 5, 10 and 20 μg ml^–1 red elemental selenium significantly induced the expression of interferon (IFN) and ISG15 genes in EPC cells. The viral TCID₅₀ (50% tissue culture infective dose) values in the EPC cells incubated with 5, 10 and 20 μg ml^–1 red elemental selenium were significantly less than those of the control. More expression of IFN and ISG15 genes and less TCID₅₀ values indicate that red elemental selenium indeed improves the antiviral capability of EPC cells. In the crucian carp fed with the food containing 5 and 10 μg g^–1 red elemental selenium, IFN expressions showed 13- and 39-fold increases at the 16th day of post-injection, and its expression was dependent on selenium concentrations. Meanwhile, no fish death occurred in all the experimental groups. In the zebrafish fed with the red worm containing 5 μg g^–1 red elemental selenium, IFN and Mx expressions and survival rate were significantly higher than those of the control. The results of this study show that red elemental selenium indeed improves the antiviral activity of fish. The antiviral effects of selenium mainly come from its immune regulation through its incorporation into selenoproteins. The optimum level of selenium contributes to improving fish immunity, whereas excess selenium causes excessive immune and inflammatory responses.     

Ultrastructure and elemental composition of the eggshell of Reeve’s pheasant ( Syrmaticus reevesii )

The eggshell of Reeve’s pheasant (Syrmaticus reevesii) collected from the Dongzhai National Nature Reserve in Henan Province, China was studied. By using scanning electron microscopy and inductively coupled plasma (ICP) spectrometry, the ultrastructure and elemental composition of the eggshell was determined. The study showed that the average thickness of the surface layer of crystals, the palisade and cone layer, and the eggshell membrane were 20.8, 220.8 and 62.5 μm, respectively, accounting for 6.8%, 72.6% and 20.6% of the total thickness of the eggshell. There were many vesicular holes in the palisade layer with an average diameter of 0.32 ± 0.08 μm (n = 30). The function of these holes might be significant to air exchange. The shape of the eggshell pore on the surface layer of crystals is round or elliptical. The fracture surface of the pore is funnel-shaped. Some granules filled the upper part of the eggshell pores. The content of 21 elements in the eggshell of wild and captive Reeve’s pheasants was compared and presented. It indicated that among the elements that made up the eggshell of the wild pheasant, the content of Ca, Mg, P and S was much higher, ω > 1 mg/g, with ω (Ca) being higher than 40% of the eggshell. The contents of Na, Si, Sr, K and Al were ω = 0.1–1 mg/g, while Fe, Zn, Pb, Mn, Cu, V and Ti had lower concentrations (ω = 1–100 μg/g). The ω of Ni, Cr, Co, Se, Cd were lower than 1 μg/g. The elemental composition in the eggshell of the captive Reeve’s pheasant kept in the Dongzhai National Natural Reserve was significantly different from that of the wild species, with a difference of over 20% on S, Cu, Fe, Al, Mn, Si, Sr, Se and Cr. The lower intake of Fe, Mn, Si and Sr on the one hand and the higher intake of S, Cu, Al and Cr on the other hand might be responsible for the low fertility of captive Reeve’s pheasants in the Dongzhai National Nature Reserve. In order to ensure that the pheasants are receiving the proper amount of nutrition and to improve their breeding success, the amount of certain elements in the food should be adjusted. Translated from Journal of Beijing Normal University (Natural Science), 2006, 42(1): 78–82 [译自: 北京师范大学学报 (自然科学版)]     

Characterization of sulfur oxidation by an autotrophic sulfur oxidizer,Thiobacillus sp. ASWW-2

An autotrophic sulfur oxidizer,Thiobacillus sp. ASWW-2, was isolated from activated sludge, and its sulfur oxidation activity was characterized.Thiobacillus sp. ASWW-2 could oxidize elemental sulfur on the broad range from pH 2 to 8. When 5–50 g/L of elemental sulfur was supplemented as a substrate, the growth and sulfur oxidation activity ofThiobacillus sp. ASWW-2 was not inhibited. The specific sulfur oxidation rate of strain ASWW-2 decreased gradually until sulfate was accumulated in medium up to 10 g/L. In the range of sulfate concentration from 10 g/L to 50 g/L, the sulfur oxidation rate could keep over 2.0 g-S/g-DCW-d. It indicated thatThiobacillus sp. ASWW-2 has tolerance to high concentration of sulfate.     

Extracellular reduction of selenite by a novel marine photosynthetic bacterium

A novel purple nonsulfur bacterium strain NKPB030619, which has resistance to over 5 mM selenite, was isolated from a marine environment. An initial concentration of 1.1 mM selenite, added to the medium, was decreased to under 0.05 mM within 5 days. The color of the cell suspension turned red within 2 days. The red coloration gradually decreased and black precipitates appeared during 2 weeks of cultivation. Under these conditions, two main types of deposit were formed extracellularly. These deposits were thought to contain red amorphous selenium and black vitreous selenium. The selenite reduction to elemental selenium in this bacterium was induced by the introduction of light and l-malic acid under anaerobic conditions. These results suggest that selenite reduction is coupled with photosynthesis and l-malic acid can serve as the indirect electron donor for its reduction. Phylogenetic analysis based on the 16S rDNA sequence showed that NKPB0360619 belongs to the α subdivision of Proteobacteria and is classified into the Rhodobacter species. The highest similarity of 86.2% was observed with R. sphaeroides. Received: 13 August 1996 / Received last revision: 6 May 1997 / Accepted: 11 May 1997     

Targeting Dinitrophenol to Mitochondria: Limitations to the Development of a Self-limiting Mitochondrial Protonophore

The protonmotive force (Δp) across the mitochondrial inner membrane drives ATP synthesis. In addition, the energy stored in Δp can be dissipated by proton leak through the inner membrane, contributing to basal metabolic rate and thermogenesis. Increasing mitochondrial proton leak pharmacologically should decrease the efficiency of oxidative phosphorylation and counteract obesity by enabling fatty acids to be oxidised with decreased ATP production. While protonophores such as 2,4-dinitrophenol (DNP) increase mitochondrial proton leak and have been used to treat obesity, a slight increase in DNP concentration above the therapeutically effective dose disrupts mitochondrial function and leads to toxicity. Therefore we set out to develop a less toxic protonophore that would increase proton leak significantly at high Δp but not at low Δp. Our design concept for a potential self-limiting protonophore was to couple the DNP moiety to the lipophilic triphenylphosphonium (TPP) cation and this was achieved by the preparation of 3-(3,5-dinitro-4-hydroxyphenyl)propyltriphenylphosphonium methanesulfonate (MitoDNP). TPP cations accumulate within mitochondria driven by the membrane potential (Δψ), the predominant component of Δp. Our hypothesis was that MitoDNP would accumulate in mitochondria at high Δψ where it would act as a protonophore, but that at lower Δψ the accumulation and uncoupling would be far less. We found that MitoDNP was extensively taken into mitochondria driven by Δψ. However MitoDNP did not uncouple mitochondria as judged by its inability to either increase respiration rate or decrease Δψ. Therefore MitoDNP did not act as a protonophore, probably because the efflux of deprotonated MitoDNP was inhibited.     

细菌还原氧化态硒产生红色单质硒的研究进展

硒是一种生命必需的微量元素,但高浓度时毒性较强且会造成环境污染。许多细菌可以将亚硒酸盐(SeO32-)或硒酸盐(SeO42-)等毒性较高的氧化态硒还原为毒性较小的红色单质硒(Se°),形成硒-蛋白复合物,它们对于获得最佳补硒方式和治理硒环境污染具有应用潜力。近年来,关于这一生物还原过程,人们进行了大量的研究,包括碳源、氧气、元素硫、谷胱甘肽以及一些氧化还原酶和膜转运蛋白等在内的多种物质都被发现可能影响或参与了细菌对硒的代谢。综述了细菌进行生物还原氧化态硒的影响因素及不同细菌产生红色单质硒机理的研究进展。     

Biotransformation of <Emphasis Type="SmallCaps">l</Emphasis>-Selenomethionine and Selenite in Rat Gut Contents

l-Selenomethionine (SeMet) and sodium selenite are widely used selenium nutritional supplements with potential benefit in preventing cancer. However, supplementation is not without risks of toxicity if intake is too high. The aim of the present study was to investigate SeMet and selenite metabolism in the gastrointestinal tract with particular focus on the formation of the volatile selenium excretion products, dimethylselenide (DMSe) and dimethyldiselenide (DMDSe). Adult male Wistar rats (n = 5) were euthanized, their intestinal tracts removed and the contents of jejunum, ileum, caecum and colon used to prepare 10% suspensions in saline. SeMet and selenite (0.5–0.6 mM) were then incubated with these suspensions at 37°C for 3 h. Caecum and colon contents were the most metabolically active towards SeMet with 30% and 15% metabolized over 3 h. DMDSe was the only volatile selenium metabolite detected accounting for 8.7 ± 1.3% of the selenium lost in caecum contents. Selenite was completely metabolized by caecum contents and 73% by colon contents under the same conditions forming DMSe (5.7 ± 0.9% of the selenium lost in caecum) and a precipitate of red amorphous elemental selenium. Based on previous literature and these results, we conclude that the gut microbiota contributes to the excretion of excess selenium through the production of methylated selenium compounds and elemental selenium.     

Blood gas parameters and the responses of erythrocytes in carp exposed to deep hypoxia and subsequent recovery

 The quantitative importance of the adrenergic response of carp erythrocytes during severe oxygen restriction is not clear at present. Quantitative differences between in vivo and in vitro studies suggest that the response of carp erythrocytes may be dependent on the actual hypoxic condition. To our knowledge, a clear picture of the blood gas status, erythrocytic responses and catecholamines measured simultaneously in carp exposed to deep severe hypoxia or anoxia has not yet been reported. Therefore, we studied the physiological response of carp exposed to deep hypoxia at 0.3 kPa and subsequent recovery. Carp were fitted with an indwelling cannula in the dorsal aorta for repeated blood sampling and the blood was analysed for hematocrit, hemoglobin, mean cellular hemoglobin content, intra- and extracellular pH, pO₂, pCO₂, total CO₂ and catecholamines. Large fluctuations in arterial pO₂ levels were observed in normoxic control carp, probably caused by the alternating breathing pattern of carp. Even at water pO₂ levels of 0.3 kPa, arterial pO₂ levels were maintained at about 0.2–0.3 kPa. Catecholamine levels were increased during deep hypoxia with noradrenaline as the predominant catecholamine. Hematological variables showed that the number of circulating erythrocytes was increased during hypoxia. The intracellular pH of carp red cells was maintained at pre-exposure values despite a considerable decrease of pH_e. In this in vivo study, a marked decrease of the proton gradient across the red cell membrane (pH_e-pH_i), as high as 0.35 pH units, was observed, which is quantitatively similar to that usually observed in salmonids during hypoxia. It is suggested that the regulation of the carp erythrocytic pH_i is probably caused to a major extent by deoxygenation of hemoglobin (Haldane effect) while adrenergic activation of the red cells is likely to contribute significantly to the observed reduction of the proton gradient. These mechanisms result in the persistence of a capacity for aerobic metabolism in carp of about 10–20% of the energy metabolism despite environmental pO₂ values of 2–3 mm Hg. Accepted: 7 May 1996     

Photosynthetic characteristics of Dunaliella salina (Chlorophyceae,Dunaliellales) in relation to β-carotene content

Photosynthetic characteristics of Dunaliella salina with high (red form) and low β-carotene (green form) concentrations were studied. D. salina growing in brine saltworks exhibited a high level of β-carotene (15 pg cell⁻¹). The rate of oxygen evolution as a function of irradiance was higher in the red than in the green form (on chlorophyll basis). Photosynthetic inhibition of the green form was observed above 500 μmol m⁻² s⁻¹. The red form appeared more resistant to high irradiance and no inhibition in O₂ evolution was observed up 2000 μmol m⁻² s⁻¹. However, when these results are expressed on a cell number basis the rate of oxygen evolution was significantly higher in the green form. Carbonic anhydrase (CA) activity (total, soluble, membrane bound) was found in red and green forms. CA was higher in the red form on a chlorophyll basis, but lower if expressed on a protein basis. The light dependent rate of oxygen evolution and photoinhibition depends on the concentration of β-carotene in D. salina cells.     

The use of amiloride to uncouple branchial sodium and proton fluxes in the brown trout, Salmo trutta

Resting proton, ammonium and sodium fluxes in Salmo trutta were 492.6 ± 19.5 (n = 29); 122.9 ± 34.2 (n = 28) and 277.1 ± 18.5 (n = 50) μmol · kg⁻¹ · h⁻¹, respectively. The resting transepithelial potential was found to be composed of three successive potentials, the outermost averaging −7.36 ± 0.19mV, the second, −14.3 ± 1.4 mV and the third −37 ± 1.7 mV. Amiloride inhibits the proton, ammonium and sodium fluxes in a dose-dependent manner at concentrations of 0.5 mmol · 1⁻¹ and 0.1 mmol · l⁻¹, but at 0.01 mmol · l⁻¹, proton and ammonium fluxes remained at control levels whilst the sodium was reduced to 70.59 ± 7.29 μmol · kg⁻¹ · h⁻¹. The trans-epithelial potential was effected in a bi-phasic manner by 0.5 mmol · l⁻¹ amiloride. An initial hyperpolarisation of ca. 6 mV was followed by a sustained depolarisation of ca. 14 mV (towards zero) which persisted until the amiloride was washed off the gill. The initial hyperpolarisation was thought to reflect a rapid inhibition of a positive inward sodium current and the subsequent depolarisation was due to the inhibition of a positive outward current (proton) which would abolish the transepithelial potential. However, at 0.01 mmol ·  l⁻¹ only the hyperpolarisation was seen, due to the inhibition of only the inward sodium current. Acetazolamide (0.1 mmol · l⁻¹) was found to have no significant effect on the proton, ammonium and sodium fluxes. These results indicate that the proton and sodium fluxes across the gill of the freshwater trout are not tightly linked. While this suggests that the trout gill resembles the model of Ehrenburg et al. (1985) of sodium uptake in frog skin, the apical potentials measured in the pavement epithelial cell(s) are too low to account for sodium uptake unless the activity of the sodium in the cells is very low. Accepted: 8 August 1996     

Comparative morphometry,biochemical and elemental composition of three marine sponges (Petrosiidae) from Gulf of Mannar,India

Abstract

Three marine sponges Neopetrosia similis, Xestospongia testudinaria and Petrosia nigricans from the Gulf of Mannar, Southeast coast of India were compared based on their morphometric parameters and their biochemical and elemental composition. These sponges showed differences inspicule : protein (spongin) ratio of 5:1, 11:1 and 13:1, respectively. Xestospongia testudinaria possessed the longest oxeae, 3.05 times than P. nigricans and 1.47 times than N. similis. Spectral analysis revealed that the spicules of these sponges are mostly composed of O (>50%) and Si (29–45%), whereas Al and Ca (4–5%) were additionally detected only in the spicules of P. nigricans. In contrast, the percentage elemental composition in pinacodermal sections was significantly different as O and Si were the maximum (23–50%); followed by Cl and Al as moderate (1–17%); and Na, S, Fe and Ca in minor quantities (0.2–6%). Cluster analysis and traditional taxonomic arrangements show that X. testudinaria and N. similis have a close relationship whereas P. nigricans was hypothesized to be their sister group. The present findings could be the key for identifying sponges in situ as well as determining whether sponges could be used to assess pollution in the sea.     

Selenium concentration in liver and kidney of free living animals (roe and red deer) from West Pomerania (Poland)

The aim of this study was to determine concentrations of selenium in the liver and kidneys of roe deer and red deer from West Pomerania, depending on the season. Altogether, samples from 169 animals were collected (96 from roe deer and 73 from red deer) in 2003–2007. The mean concentration of selenium in the liver of red deer and roe deer was 0.37 μg/g and 0.62 μg/g dry weight, respectively. In kidneys, Se concentration was 2.72 μg/g d.w. in red deer and 2.99 μg/g d.w. in roe deer. In roe deer, liver selenium concentration in autumn was significantly higher than in winter (P < 0.05) and spring (P < 0.01) and significantly lower in spring than in summer (P < 0.05); likewise, kidney selenium concentration was higher in autumn than in summer. In deer, no statistically significant season-related differences were observed for liver selenium concentrations. In red deer kidneys, selenium concentration was the lowest in summer, significantly lower than in autumn and winter. Low selenium concentrations in the analyzed tissues show that the animals live in areas deficient in this element.     

A mathematical model for drug administration by using the phagocytosis of red blood cells

 A mathematical model for the delivery of drug directly to the macrophages by using the phagocytosis of senescent red blood cells is proposed. The model is based on the following assumption: At time t=0 a preassigned red blood cell population n(0, a)=φ(a), a>0, loaded by the drug, is injected in the blood circulation. Among the cells of that population only those with an age a≧ā (ā=120 days) will be phagocytosed by macrophages. Of course, the lifetime of the drug must be higher than ā. Within the red blood cells it cannot be metabolized, neither can it diffuse through their membranes. The emphasis of the paper is on the mathematical properties and on the formulation of the control problem. Received 15 December 1994; received in revised form 20 July 1995     

Raman microspectrometry sulfur detection and characterization in the marine ectosymbiotic nematode Eubostrichus dianae (Desmodoridae, Stilbonematidae)

Background information. Marine nematodes belonging to the Stilbonematidae (Desmodoridae) family are described as living in obligatory association with sulfur‐oxidizing chemoautotrophic ectosymbionts. The symbiotic bacteria carrying out this chemosynthesis should contain elemental sulfur in periplasmic granules as sulfur granules of chemoautotrophic endosymbionts described in various marine invertebrates. Results. Based on TEM (transmission electron microscopy) analyses, extracellular bacteria surrounding Eubostrichus dianae possess these spherical periplasmic granules. Few investigative techniques can be used to identify elemental sulfur, S₈, such as EDXS (energy dispersive X‐ray spectroscopy) and EELS (electron energy loss spectroscopy), which are associated with cryo‐fixation of the sample to avoid sulfur loss. These techniques are time consuming, expensive and require technical skills. Raman microspectrometry applied to the analysis of E. dianae allowed us to detect elemental sulfur, S₈, and confirmed the location of these sulfur clusters in the bacterial coat. In the same way, Raman spectrometry was positively applied to the endosymbiotic bivalve Codakia orbicularis, suggesting that this technique can be used to characterize sulfur in ecto‐ as well as in endo‐symbiotic sulfur‐oxidizing bacteria. Conclusions. As Raman spectrometry can be used on living organisms (without preliminary fixation) without sample damage and preserving the molecular structure of the sulfur (denatured during chemical fixation), it represents a very well‐adapted investigative tool for biologists. This technique therefore permits us to detect quickly and easily (in a few seconds and on entire living animals) the presence of sulfur compounds in the symbiotic nematode.     

pH-Induced Proton Permeability Changes of Plasma Membrane Vesicles

In vivo studies with leaf cells of aquatic plant species such as Elodea nuttallii revealed the proton permeability and conductance of the plasma membrane to be strongly pH dependent. The question was posed if similar pH dependent permeability changes also occur in isolated plasma membrane vesicles. Here we report the use of acridine orange to quantify passive proton fluxes. Right-side out vesicles were exposed to pH jumps. From the decay of the applied ΔpH the proton fluxes and proton permeability coefficients (P_H+) were calculated. As in the intact Elodea plasma membrane, the proton permeability of the vesicle membrane is pH sensitive, an effect of internal pH as well as external pH on P_H+ was observed. Under near symmetric conditions, i.e., zero electrical potential and zero ΔpH, P_H+ increased from 65 × 10⁻⁸ at pH 8.5 to 10⁻¹ m/sec at pH 11 and the conductance from 13 × 10⁻⁶ to 30 × 10⁻⁴ S/m². At a constant pH _i of 8 and a pH _o going from 8.5 to 11, P_H+ increased more than tenfold from 2 to 26 × 10⁻⁶ m/sec. The calculated values of P_H+ were several orders of magnitude lower than those obtained from studies on intact leaves. Apparently, in plasma membrane purified vesicles the transport system responsible for the observed high proton permeability in vivo is either (partly) inactive or lost during the procedure of vesicle preparation. The residue proton permeability is in agreement with values found for liposome or planar lipid bilayer membranes, suggesting that it reflects an intrinsic permeability of the phospholipid bilayer to protons. Possible implications of these findings for transport studies on similar vesicle systems are discussed. Received: 5 April 1995/Revised: 28 March 1996     

Geranylhydroquinone 3′′-hydroxylase, a cytochrome P-450 monooxygenase from Lithospermum erythrorhizon cell suspension cultures

Geranylhydroquinone 3′′-hydroxylase, which is likely to be involved in shikonin and dihydroechinofuran biosynthesis, was identified in cell suspension cultures of Lithospermum erythrorhizon Sieb. et Zucc. (Boraginaceae). The enzyme hydroxylates the isoprenoid side chain of geranylhydroquinone (GHQ), a known precursor of shikonin. Proton/proton correlation spectroscopic and proton/proton long-range correlation spectroscopic studies confirmed that hydroxylation takes place specifically at position 3′′, i.e. at the methyl group involved in the cyclization reaction. The enzyme is membrane-bound and was found in the microsomal fraction. It requires NADPH and molecular oxygen as cofactors, and is inhibited by cytochrome P-450 inhibitors such as cytochrome c and CO. The inhibitory effect of CO is reversed by illumination. These data suggest that the enzyme is a cytochrome P-450-dependent monooxygenase. The optimum pH of GHQ 3′′-hydroxylase is 7.4, and the apparent K _m value for GHQ is 1.5 μM. The reaction velocity obtained with 3-geranyl-4-hydroxybenzoic acid was more than 100 times lower than that obtained with geranylhydroquinone. Received: 20 March 1999 / Accepted: 20 July 1999     

Reduction of Selenite to Elemental Red Selenium by <Emphasis Type="Italic">Rhizobium</Emphasis> sp. Strain B1

A bacterium that reduces the soluble and toxic selenite anion to insoluble elemental red selenium (Se⁰) was isolated from a laboratory bioreactor. Biochemical, morphological, and 16S rRNA gene sequence alignment identified the isolate as a Rhizobium sp. that is related to but is genetically divergent from R. radiobacter (syn. Agrobacterium tumefaciens) or R. rubi (syn. A. rubi). The isolate was capable of denitrification and reduced selenite to Se⁰ under aerobic and denitrifying conditions. It did not reduce selenate and did not use selenite or selenate as terminal e⁻ donors. Native gel electrophoresis revealed two bands, corresponding to molecular weights of ∼100 and ∼45 kDa, that reduced selenite. Tungsten inhibited in vivo selenite reduction, suggesting that a molybdenum-containing protein is involved in selenite reduction. This organism, or its enzymes or DNA, might be useful in bioreactors designed to remove selenite from water.     

Mineral composition variation in the shells of freshwater mussel Anodonta woodiana at different growth stages

The levels of mineral element Na, Mg, K, Ca, Cr, Mn, Fe, Co, Ni, Cu, Zn, Se, Mo, Al, As, Ag, Cd, and Tl were quantified in the whole shells of the freshwater bivalve Anodonta woodiana at three different growth stages (i.e. J1 juveniles of 1 month old, J2 juveniles of 3.5 months old, and adults of 36 months old). The concentrations of Na and Al were different between different growth stages (p < 0.05). The highest Na concentrations (2715 ± 86 μg/g dry weight) were found in J2 juveniles. The highest Al concentrations (303.9 ± 5.95 μg/g dry weight) were found in J1 juveniles. Manganese concentrations (517.0 ± 47.98 μg/g dry weight) were significantly higher in J2 juveniles than in J1 juveniles (432.3 ± 9.87 μg/g dry weight) (p < 0.05). Copper concentrations (27.32 ± 0.15 μg/g dry weight) were significantly higher in J1 juveniles than in J2 juveniles (26.21 ± 0.86 μg/g dry weight) and adults (24.74 ± 1.43 μg/g dry weight) (p < 0.05). Burdens of Na, Ca, Mn, Fe, Co, Cu, Mo, Ag, and Tl were positively correlated with the shell length (p < 0.05). These findings can possibly contribute to an understanding of elemental requirements for shell growth and, hence, facilitate improvement of survival and growth rates during artificial mussel culture.     

Micro‐particle‐induced X‐ray emission mapping of elemental distribution in roots of a Mediterranean‐type sclerophyll,Agathosma betulina (Berg.) Pillans,colonized by Cryptococcus laurentii

The role of rhizosphere yeasts as plant nutrient‐scavenging microsymbionts in resource‐limited Mediterranean‐type heathlands is unknown. This study, therefore, focused on quantitative elemental distribution within the roots of a medicinal sclerophyll, Agathosma betulina (Berg.) Pillans, grown under nutrient‐poor conditions, and colonized by Cryptococcus laurentii. Micro‐particle‐induced X‐ray emission (PIXE) was used to assess quantitative elemental distribution within the roots of A. betulina inoculated with viable C. laurentii, as well as within roots of control plants that received autoclaved yeast. To aid in the interpretation of heterogeneous elemental distribution patterns, apoplastic barriers (Casparian bands) in root tissues were located using fluorescence microscopy. In addition, root cross‐sections were examined for endophytic C. laurentii using light and transmission electron microscopy (TEM). The average concentrations of P, Fe and Mn were significantly (P < 0.05) higher in roots of yeast‐inoculated plants, compared to control plants. Casparian bands were observed in the exodermal cells of both treatments, and the presence of these bands was correlated with elemental enrichment in the epi/exodermal‐outer cortical tissues. Light and TEM micrographs revealed that the yeast was not a root endophyte. This is the first report describing the role of a soil yeast as a plant nutrient‐scavenging microsymbiont.     

Relationship between rates of respiratory proton extrusion and ATP synthesis in obligately alkaliphilic Bacillus clarkii DSM 8720T

To elucidate the energy production mechanism of alkaliphiles, the relationship between the rate of proton extrusion via the respiratory chain and the corresponding ATP synthesis rate was examined in obligately alkaliphilic Bacillus clarkii DSM 8720^T and neutralophilic Bacillus subtilis IAM 1026. The oxygen consumption rate of B. subtilis IAM 1026 cells at pH 7 was approximately 2.5 times higher than that of B. clarkii DSM 8720^T cells at pH 10. The H⁺/O ratio of B. clarkii DSM 8720^T cells was approximately 1.8 times higher than that of B. subtilis IAM 1026 cells. On the basis of oxygen consumption rate and H⁺/O ratio, the rate of proton translocation via the respiratory chain in B. subtilis IAM 1026 is expected to be approximately 1.4 times higher than that in B. clarkii DSM 8720^T. Conversely, the rate of ATP synthesis in B. clarkii DSM 8720^T at pH 10 was approximately 7.5 times higher than that in B. subtilis IAM 1026 at pH 7. It can be predicted that the difference in rate of ATP synthesis is due to the effect of transmembrane electrical potential (Δψ) on protons translocated via the respiratory chain. The Δψ values of B. clarkii DSM 8720^T and B. subtilis IAM 1026 were estimated as −192 mV (pH 10) and −122 mV (pH 7), respectively. It is considered that the discrepancy between the rates of proton translocation and ATP synthesis between the strains used in this study is due to the difference in ATP production efficiency per translocated proton between the two strains caused by the difference in Δψ.