Electrophysiological Evidence for an Electrogenic Proton Pump and the Proton Symport of Glucose in the Marine Fungus Dendryphiella salina

The marine hyphomycete Dendryphiella salina (Suth.) Nicot &Pugh has a resting membrane potential of –250 mV (insidenegative). The respiratory inhibitors sodium azide and FCCPinduced a rapid but reversible depolarization of the membraneof at least 180 mV; sodium azide also caused alkalinizationof the medium. Vanadate brought about significant depolarizationbut this was not always reversible. EDTA induced depolarizationthough to a lesser extent. DIDS and SITS caused a depolarizationof around 30–70 mV which was readily reversible, N-ethylmaleimideirreversibly depolarized the membrane by 180–200 mV. Ouabainhad no effect. When external concentrations of H⁺ , K⁺ , Na⁺or Cl^– were changed singly, only changes in H⁺ affectedmembrane potential, with shifts decreasing with increasing pH.Glucose and 3-O-methyl glucose depolarized the membrane in aconcentration-dependent manner which was enhanced by starvationof the hyphae. Recovery occurred in the presence of the hexose.Glucose caused an alkalinization of the medium, with time characteristicssimilar to the membrane potential changes. It is concluded thatthere is an electrogenic proton pump and a proton—glucosesymporter in D. salina. The retention of proton-based transportsystems suggests a terrestrial origin for the fungus. Key words: Marine fungi, Dendryphiella salina, membrane potential, electrogenic proton pump, proton symport, hexose     

X-ray Microanalysis of Ion Distribution in Abutilon Nectary Hairs

Robards, A. W. and Oates, K. 1986. X-ray microanalysis of iondistribution in Abutilon nectaryhairs—J. exp. Bot. 37:940–946. X-ray microanalysis of frozen-hydrated Abutilon nectary hairsshows that potassium is present at easily detectable levelsin all cells between the phloem and the trichome tip cells.There is little overall variation in the amount of potassiumfrom cell to cell although it was found that: (i) some basalcells accumulate potassium to a significantly higher level thanthe hair cells; and (ii) there is more potassium in the cellstowards the distal end of the trichome. These findings wouldbe consistent with a model for secretion envisaging a trans-membraneloading of nectar into a ‘secretory reticulum’ inall trichome cells. This loading process would be selectiveand would exclude ions. A pronounced increase in the level ofchlorine from the basal cell along the hair was observed. Nocomplementary cation was detected but equivalent changes inthe level of Na would have been below the detection limit ofthe system. Key words: Abutilon, nectary hairs, X-ray microanalysis, ion distribution     

Subcellular Adaptation to Salinity and Irradiance in Dunaliella salina

Dunaliella salina V-63 was cultivated in different concentrations of NaCl (0.5, 1.0, 2.5, 3.0, or 4.0 M) and at two irradiances (170 or 220 μmol m⁻²s⁻¹). Concentration-dependent suppression of growth was observed above 1 M NaCl, and elevated salinity induced formation of salt-containing vacuoles. However, the changes in the chloroplast ultrastructure following changes in salinity and irradiance (increase of invaginations and protuberances, numerous grana with low number of thylakoids, less number of starch grains, etc.) appeared to be of primary importance. This revised version was published online in July 2006 with corrections to the Cover Date.     

Lipid Composition of the Marine Ciliates Pleuronema sp. and Fabrea salina: Shifts in Response to Changes in Diet1

ABSTRACT. The composition and incorporation of lipids in two marine ciliates, Pleuronema sp. and Fabrea salina, was examined following growth on either an algal or bacterial diet. When allowed to feed on a natural bacterial community, Pleuronema sp. synthesized the triterpenoid alcohol gammaceran-3β-ol (tetrahymanol) and two hopanoids (hopan-3β-ol and one uncharacterized hopanoid). When fed the marine alga Isochrysis galbana, F. salina contained the major algal sterol 24-methylcholesta-5, 22-dien-3β-ol and several long chain ketones specific to the alga. In both ciliates, fatty acids composition showed a general correspondence to that of the diet. Using a series of antibiotic treatments to alter the bacterial prey community, and thus fatty acid composition of the ciliate's diet, promoted changes in the fatty acid composition of Pleuronema sp. to resemble that of the bacterial prey. The addition of a mixture of algal sterols to a bacterized culture of another scuticociliate, Parauronema acutum, inhibited tetrahymanol synthesis and resulted in the incorporation of sterols into the ciliate.     

A Comparison of the Methods of X-ray Microanalysis, Compartmental Analysis and Longitudional Ion Profiles to Estimate Cytoplasmic Ion Concentrations in Two Maize Varieties

Hajibagheri, M. A., Flowers, T. J., Collins, J. C. and Yeo,A. R. 1988. A comparison of the methods of X-ray microanalysis,compartmental analysis and longitudinal ion profiles to estimatecytoplasnuc ion concentrations in two maize varieties.—J.exp. Bot. 39: 279-290. The ion content of compartments within plant root cells hasbeen studied by three different methods; flux analysis usingradioactive isotopes, longitudinal ion profiles and X-ray microanalysis.The data provide estimates of the concentrations of K⁺ and CI^–in the cytoplasm of roots of culture solution and salt grownmaize by three independent methods. In the cultivar LG₁₁ grown in 50 mol m^–3 NaCl X-ray microanalysis,compartmental analysis and longitudinal profiles yielded approximatelyagreeing values for cytoplasmic K⁺ of 90, 70 and 62 mol m^–33of tissue volume respectively. However, the methods disagreedon cytoplasmic Cl^– where the value obtained by compartmentalanalysis was about four times that from X-ray microanalysisand longitudinal profiles. Possible reasons for this are discussed. Key words: X-ray microanalysis, longitudinal ion profiles, compartmental analysis, salt-tolerance.     

盐生杜氏藻对盐度改变的生理响应

以盐生杜氏藻为实验材料,采用f/2培养基,设置了8个盐度(15、20、25、30、50、70、90、110)处理,分盐度改变前(A)和盐度改变后(B)两个实验阶段,研究了盐生杜氏藻在不同盐度处理下的生长情况,测定了藻液的OD值、叶绿素a、β-胡萝卜素、可溶性蛋白质和可溶性糖含量等指标。结果表明,A阶段,几个较低盐度(15、20、25和30)处理生长状况较好,其中又以盐度20的处理最好;余下的处理,盐度越高,其生长所受的影响越大。B阶段,盐生杜氏藻的生长进入平台期后,50、70、90、110几个盐度较高处理的细胞密度、叶绿素a、β-胡萝卜素含量均显著超过了作为对照的盐度20的处理。且B阶段末期,先前盐度15的处理蛋白质、糖的积累量,与A阶段末期相比都有了不同程度的增加,而其余盐度处理组的蛋白质、糖含量则分别产生了不同程度的下降。     

Intercellular Compartmentation of Ions in Barley Leaves in Relation to Potassium Nutrition and Salinity

X-ray microanalysis was used to determine the distributionsof several nutrient elements between vacuoles of epidermal andmesophyll cells in barley leaves and these distributions wererelated to shoot nutrient concentrations. Under the growth conditionsused, P was found only in mesophyll vacuoles, never in the epidermis.In contrast, Cl and Ca were located almost exclusively in theepidermis while K and Na were more evenly distributed betweenthe two cell types. The compartmentation of Ca and Cl in theepidermis was maintained over a wide range of tissue concentrationsof these ions. In particular, Cl was excluded from the mesophyllof salt-grown barley until the tissue concentration reachedabout 170 mol m^–3 and then it appeared in the vacuolesof these cells, but only at low concentrations. In contrast,Na was not excluded from the mesophyll of salt-grown or K-deficientbarley and there was evidence that this ion was preferentiallyaccumulated in the mesophyll. Nutrients were evenly distributedbetween the adaxial and abaxial epidermal layers, except K.which was at slightly higher concentrations in the adaxial epidermis.There was considerable variation in the concentrations of ionsin adjacent epidermal cells. The results indicate that intercellularcompartmentation of nutrients occurs in barley leaves and therole of this phenomenon in responses to nutrient deficienciesand salinity is discussed Key words: Compartmentation, barley, salinity, ions, potassium nutrition     

Phototaxis in the Marine Fungus Rhizophydium littoreum

Phototaxis appears to be a factor that influences the dispersal of zoospores of the marine fungus Rhizophydium littoreum. By using a quantitative method to study phototaxis, zoospores were found to be positively phototactic toward blue wavelengths of light. White light in the range of 20 to 6,000 microeinsteins m⁻² s⁻¹ and blue light in the range of 1 to 300 microeinsteins m⁻² s⁻¹ gave positive responses in the laboratory. Results of field studies confirmed the ability of zoospores to respond to light under natural conditions. Phototaxis may be an important adaptive mechanism which influences vertical migration of zoospores in the photic zone where plant products are available as nutrients.     

Chemotaxis in the Marine Fungus Rhizophydium littoreum

Zoospores of the marine chytrid Rhizophydium littoreum are attracted to a variety of substances common to their environment. In general, carbohydrates and polysaccharides elicited strong concentration-dependent positive responses. There was no direct correlation between all substances used as foods and those stimulating positive responses. The chemotactic activities of this organism should, however, tend to bring it toward concentrated food sources.     

Determination of Ion Content and Ion Fluxes in the Halotolerant Alga Dunaliella salina

A method to determine intracellular cation contents in Dunaliella by separation on cation-exchange minicolumns is described. The separation efficiency of cells from extracellular cations is over 99.9%; the procedure causes no apparent perturbation to the cells and can be applied to measure both fluxes and internal content of any desired cation. Using this technique it is demonstrated that the intracellular averaged Na⁺, K⁺, and Ca²⁺ concentrations in Dunaliella salina cultured at 1 to 4 molar NaCl, 5 millimolar K⁺, and 0.3 millimolar Ca²⁺ are 20 to 100 millimolar, 150 to 250 millimolar, and 1 to 3 millimolar, respectively. The intracellular K⁺ concentration is maintained constant over a wide range of media K⁺ concentrations (0.5-10 millimolar), leading to a ratio of K⁺ in the cells to K⁺ in the medium of 10 to 1,000. Severe limitation of external K⁺, induces loss of K⁺ and increase in Na⁺ inside the cells. The results suggest that Dunaliella cells possess efficient mechanisms to eliminate Na⁺ and accumulate K⁺ and that intracellular Na⁺ and K⁺ concentrations are carefully regulated. The contribution of the intracellular Na⁺ and K⁺ salts to the total osmotic pressure of cells grown at 1 to 4 molar NaCl, is 5 to 20%.     

猫肾离体保存中亚细胞水平上Ca^2＋浓度的X—射线微区分析

应用定量Ｘ－射线微区分析技术结合细胞化学技术,分析测定用单纯冷冻法保存离体猫肾脏过程中肾脏细胞的胞浆、线粒体、内质网、细胞核等细胞器内的Ｃａ２＋浓度变化,并探索钙通道阻滞剂对这种变化的影响。保存３６小时及７２小时后,线粒体与胞浆中Ｃａ２＋的峰背比极显著地提高,内质网、细胞核中钙颗粒减少。添加Ｖｅｒａｐａｍｉｌ后,保存过程中细胞器内Ｃａ２＋无显著变化。线粒体中的Ｃａ２＋峰背比与胞浆中的呈强的正相关,ｒ＝０９９０。实验结果显示：保存过程中,Ｃａ２＋由钙库（内质网等）进入胞浆中,线粒体在胞浆Ｃａ２＋浓度高时摄取Ｃａ２＋,而钙通道阻滞剂可抑制该过程。     

The relationship between the uptake of glucose and 3-O-methylglucose and soluble carbohydrate and polysaccharide in the fungus Dendryphiella salina.

When mycelium of Dendryphiella salina, pre-incubated in D-[I-14C]mannitol such that this is the only major labelled soluble carbohydrate present, absorbs glucose or the non-metabolized sugar 3-O-methylglucose, there is a specific stimulation of incorporation of 14C into alkali-insoluble, trichloroacetic-acid-soluble (1 leads to 4)-alpha-glucan, probably glycogen. There is also a net increase in the amount of glucan caused by stimulation of synthesis and inhibition of breakdown. Addition of 3-O-methylglucose results in the loss of mannitol into the medium. This loss and the reduced rate of replenishment from the glucan is important in the osmotic regulation of the hyphae. If osmotic adjustment does not occur, the hyphae do not show the specific incorporation of 14C into the glucan.     

The Effects of Salinity upon Ion Content and Ion Transport of the Marine RedAlga Porphyra purpurea(Roth) C.Ag.

Ion contents and concentrations (K⁺, Na⁺, Cl^–, Ca²⁺, Mg²⁺,SO^2–₄, NO^–₃, HPOJ^2–₄, amino and organic acids)of P. purpurea have been studied in relation to salinity variation.Cells were shown to accumulate large amounts of K⁺ and Cl^–against their respective gradients of electrochemical potentialin all dilute and concentrated seawater media. Active influxof SOJ^2–₄, NO^–₃, and HPOJ^2–₄ is also suggested,while Na⁺ is actively excluded from cells under hyposaline andhypersaline conditions. The relative proportions of individualcomponents of the internal osmotic potential were found to changeaccording to the external salt concentration. KCL forms themajor fraction of _j} in concentrated seawater media while K⁺-aminoacids form the major fraction in dilute seawaters. Other intracellularsolutes comprise less than 15% oft_j, in all media. Unidirectional fluxes of K⁺ and Cl^– were studied by radioisotopicmeans. Fluxes of K⁺ and Cl^– are reduced in hyposalinemedia, as is absolute KCL content per cell. Intracelhilar KCLcontent was also found to be markedly dependent upon externalK⁺ concentration, rather than water potential. Changes in KC1levels induced by salinity variation occur over a 6 h period.     

The Physiological Response of Synechococcus elongatus to Salinity: A Potential Biomarker for Ancient Salinity in Evaporative Environments

The purpose of this study was to characterize the physiological response of Synechococcus elongatus, a brackish-water cyanobacterium, to salt stress. S. elongatus was grown in artificial sea water medium with different salinities. The response was measured by analysis of extracellular polymeric substances (EPS) and membrane lipids. The EPS yields were positively correlated (r² = 0.99) with the salinity. The ratio of unsaturated to saturated fatty acids (U/S) increased with salinity in the range of 2.1‰ to 31.5‰ and decreased at 52.5‰. A positive linear correlation (r² = 0.92) was observed between the average chain length (ACL) of fatty acids and the salinity. These data indicate that S. elongatus adapted to salt stress by the secretion of EPS and by adjusting the membrane fluidity through the changes in ACL or desaturation of fatty acids. These variations in EPS and fatty acids may be used as geochemical biomarkers in sediments to unravel changes in the salinity of ancient evaporative environments.     

Kinetics of the Anti-oxidant Response to Salinity in the Halophyte Cakile maritima

The effects of NaCl stress on the activity of anti-oxidant enzymes (superoxide dismutase, catalase (CAT),peroxidase (POD),ascorbate peroxidase (APX), monodehydroascorbate reductase, dehydroascorbate reductase (DHAR), and glutathionereductase (GR)), anti-oxidant molecules (ascorbate and glutathione), and parameters of oxidative stress (malondialdehyde(MDA), electrolyte leakage, and H_2O_2 concentrations) were investigated in Cakile maritima, a halophyte frequent along theTunisian seashore. Seedlings were grown in the presence of salt (100, 200, and 400 mmol/L NaCI). Plants were harvestedperiodically over 20 days. Growth was maximal in the presence of 0-100 mmol/L NaCl. At 400 mmol/L NaCl, growthdecreased significantly. The salt tolerance of C. maritima, at moderate salinities, was associated with the lowest values ofthe parameters indicative of oxidative stress, namely the highest activities of POD, CAT, APX, DHAR, and GR and high tissuecontent of ascorbate and glutathione. However, prolonged exposure to high salinity resulted in a decrease in anti-oxidantactivities and high MDA content, electrolyte leakage, and H_2O_2 concentrations. These results suggest that anti-oxidantsystems participate in the tolerance of C. maritima to moderate salinities.     

Kinetics of the Anti-oxidant Response to Salinity in the Halophyte Cakile maritima

The effects of NaCl stress on the activity of anti-oxidant enzymes （superoxide dismutase, catalase （CAT）, peroxidase （POD）, ascorbate peroxidase （APX）, monodehydroascorbate reductase, dehydroascorbate reductase （DHAR）, and glutathione reductase （GR））, anti-oxidant molecules （ascorbate and glutathione）, and parameters of oxidative stress （malondialdehyde （MDA）, electrolyte leakage, and H2O2 concentrations） were investigated in Cakile maritima, a halophyte frequent along the Tunisian seashore. Seedlings were grown in the presence of salt （100, 200, and 400 mmol/L NaCl）. Plants were harvested periodically over 20 days. Growth was maximal in the presence of 0-100 mmol/L NaCl. At 400 mmol/L NaCl, growth decreased significantly. The salt tolerance of C. maritima, at moderate salinities, was associated with the lowest values of the parameters indicative of oxidative stress, namely the highest activities of POD, CAT, APX, DHAR, and GR and high tissue content of ascorbate and glutathione. However, prolonged exposure to high salinity resulted in a decrease in anti-oxidant activities and high MDA content, electrolyte leakage, and H2O2 concentrations. These results suggest that anti-oxidant systems participate in the tolerance of C. maritima to moderate salinities.     

Radiolabelling Studies of Lipids in the Marine Cryptomonad Chroomonas salina in Relation to Fatty Acid Desaturation

Cells of Chroomonas salina were exposed to [¹⁴C]acetate, [^l4C]16:0,[¹⁴C]18:0, [¹⁴C]18:1(n-9), [¹⁴C]18:2(n–6) or [¹⁴C]18:3(n–3)for 1 h and then incubated for 24 h in non-radioactive medium.At the end of the pulse period, non-glycolipid polar lipidscontained the highest proportions of radioactivity incorporatedfrom [¹⁴C]acetate and [¹⁴C]18:3(n–3) whereas with [¹⁴C]16:0,[¹⁴C]18:1 and [¹⁴C]18:2(n–6), triacylglycerols were mosthighly labelled. ¹⁴C-18:0 was recovered mainly as non-esterifiedfatty acid. Monogalactosyldiacylglycerol initially contained17% of label incorporated from [¹⁴C]acetate but less than 3%of that from [¹⁴C]fatty acids. With all substrates, excluding[¹⁴C]18:0, a gradual transfer of label from polar lipids totriacylglycerols was observed during the chase period. Saturatesand monoenes synthesised from [¹⁴C]acetate were mostly transferedfrom phospholipids and glycolipids to neutral lipid withoutfurther desaturation. Most of the incorporated ¹⁴C-fatty acidsremained unchanged and only with [¹⁴C]18:3(n–3) was substantialamounts of label recovered in penta- and hexaenoic fatty acids.The results indicate that, under the conditions of the study,lipid synthesis in the algae was heavily dominated by triacylglycerolformation and that the mechanisms of fatty acid desaturationin this species may differ from those in higher plants. (Received December 10, 1991; Accepted March 6, 1992)     

X-ray Microanalysis of the Seminal Root of Sorghum bicolor with Particular Reference to Silicon

Sorghum bicolor (L.) Moench. cv. P508.GB plants were grown inwater culture for 1 week, when the seminal roots were harvestedand sampled at five positions starting from the base: 0.0, 0.25,0.50 and 0.75 of the axis length, and a sub-apical position,11 mm behind the tip. Mineral distribution in bulk frozen rootsegments was investigated using SEM and X-ray microanalysis.The elements detected were potassium, chlorine, sulphur, sodium,phosphorus, calcium and silicon. The first four occurred inall root zones. Phosphorus was ubiquitous, but appeared to accumulatein the pericycle protoplasm. Calcium and silicon exhibited themost variation along the seminal axis. Calcium was present inall tissues at the base, but decreased acropetally, being detectedin only the outer cortical and epidermal walls of the subapicalzone. Silicon was present at low levels in protoplasts and wallsof most root tissues, and accumulated in the endodermal protoplastand walls. Deposition in walls is initiated coincident withthe earliest stages of secondary wall thickening. Silicon contentof the inner tangential wall of the endodermis exhibits a decreasingacropetal gradient along the axis length. It is absent frommost cell walls of the sub-apical zone. Silicon pathways inthe root, and silica aggregate formation in relation to thesurrounding ionic environment, are discussed. Sorghum bicolor (L). Moench, seminal root, cryostage, SEM, X-ray microanalysis, ion localization, silicon, endodermis