Salt stress affects sterol biosynthesis in the halophilic black yeast Hortaea werneckii

Hortaea werneckii is a black yeast recently isolated from salterns in Slovenia. Some of the adaptations of halophilic microorganisms to increased salinity and osmolarity of the environment are alterations in membrane properties. By modulating the fluidity, sterols play an important role as a component of eukaryotic biological membranes. We studied the regulation of sterol biosynthesis in H. werneckii through the activity and amount of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG R), a key regulatory enzyme in the biosynthesis of sterols. We found some differences in the characteristics of HMG R and in its regulation by different environmental salinities in H. werneckii when compared to the mesophilic baker's yeast, Saccharomyces cerevisiae. Our results suggest that halophilic black yeast regulates sterol biosynthesis through HMG R in a different way than mesophiles, which might be a consequence of the different ecophysiology of halophilic black yeasts. From this perspective, H. werneckii is an interesting novel model organism for studies on salt stress-responsive proteins as well as on sterol biosynthesis in eukaryotes.     

Hypersaline waters in salterns - natural ecological niches for halophilic black yeasts

Hypersaline waters in salterns have so far been considered to be populated only with halophilic algae and bacteria and completely lacking halophilic fungi. In this paper we present population dynamics of polymorphic black yeasts, isolated from hypersaline waters (3-30% NaCl) of a saltern, in relation to different physicochemical parameters. Hortaea werneckii, Phaeotheca triangularis, Trimmatostroma salinum, Aureobasidium pullulans and Cladosporium spp. were detected with the highest frequency just before the peak of halite (NaCl) concentration. Since H. werneckii, P. triangularis and T. salinum are not known outside saline environments, these results suggest that hypersaline water is their natural ecological niche.     

Identity and biodegradative abilities of yeasts isolated from plants growing in an arid climate

Plants harvested in the Canary Islands Lanzarote and Fuerteventura were analyzed for the yeasts inhabiting their surface. Half of the isolates (22 out of 44) were identified as Debaryomyces hansenii. Black ascomycetes, viz. Hortaea werneckii and two Hormonema species were represented by 7 strains. Basidiomycetous yeasts, viz. Cryptococcus sp. (8 strains), Rhodotorula sp. (5 strains), Cerinosterus cyanescens (1 strain) and Pseudozyma sp. (1 strain) constituted a minority of 33%. Thirty strains were screened for their ability to assimilate various plant constituents including lipids of the cuticle and the cell membrane, hemicelluloses, nitrogenous compounds (protein, nucleic acids, amino acids) and benzene compounds. All strains were able to assimilate or to hydrolyze lipids, lecithin included. Many strains of D. hansenii, H. dematioides, H. werneckii, C. cyanescens, Cr. laurentii, Pseudozyma sp. and Rh. glutinis were proteolytic. Hemicelluloses like xylan and pectin were assimilated by black ascomycetous yeasts, Cryptococcus sp., Pseudozyma sp. and Rh. glutinis. Ferulic and hydroxycinnamic acids, gallic and tannic acids were assimilated by some strains of H. dematioides, C. cyanescens, Pseudozyma sp. and Rhodotorula sp.     

Properties of native and solubilized plasma membrane ATPase from the halophyte Plantago crassifolia, grown under saline and non-saline conditions

Plasma membrane ATPase from the mediterranean halophyte Plantago crassifolia Forskal was analysed in the native and solubilized state. The enzyme revealed a broad pH optimum at 6.3 when analysed in plasma membrane preparations from roots. A further purified preparation of leaf plasma membrane ATPase was obtained by a three-step solubilization method. Final solubilization of the enzyme was achieved with 1% lysophosphatidylcholine. This enzyme showed a narrow pH optimum at 6.3, inhibition by vanadate, fluoride and N,N'-dicyclohexylcarbodiimide and a high specificity for ATP. The preparations contained a major polypeptide of 107 kDa. None of the parameters analysed in this enzyme changed upon transfer to saline conditions, although the leaves accumulated Na⁺ and Cl⁻ and an enhanced formation of the compatible organic osmoticum, sorbitol, was detected. It is concluded that at least in this halophyte, the plasma membrane ATPase undergoes no changes during the physiological adaptation of the plant to a saline environment.     

出芽短梗霉在三种不同氮源的培养基中多糖积累以及形态变化

本研究旨在阐明出芽短梗霉在不同氮源培养基中形态和胞外多糖的积累及化学成分变化。采用摇瓶法培养出芽短梗霉。三种培养基的氮源分别为硝酸钠(培养基1,M1)、硫酸氨、酵母膏(培养基2,M2)和硫酸氨、蛋白胨和酵母膏(培养基3,M3)。M1培养基中,菌丝体和单细胞的生物量积累均比M2、M3低,但胞外多糖的产量则等于甚至略超过M2和M3。在指数生长的前期,白色菌丝体和酵母状细胞状态占优势。指数生长的后期,以厚垣孢子、肿大细胞和黑色菌丝体占优势。胞外多糖都能为茁霉多糖酶水解为麦芽糖和麦芽三糖,说明这些多糖的化学组成都具有(1→4,1→6)-α结构的茁霉多糖。但M1中产生的茁霉多糖结构单元为麦芽糖和麦芽三糖,且二者比例相当。M2中茁霉多糖的麦芽糖结构单元明显减少,而M3中144h后麦芽糖结构单元完全消失。这似乎表明氧化性的氮源和低溶解氧水平可能是造成茁霉多糖结构单元同时具有麦芽糖和麦芽三糖的原因。     

Microbial community biomass and structure in saline and non-saline soils associated with salt- and boron-tolerant poplar clones grown for the phytoremediation of selenium

Poplar trees (Populus spp.) are often used in bioremediation strategies because of their ability to phytoextract potential toxic ions, e.g., selenium (Se) from poor quality soils. Soil microorganisms may play a vital role in sustaining health of soil and/or tolerance of these trees grown in poor quality soils by contributing to nutrient cycling, soil structure, overall soil quality, and plant survival. The effect of naturally occurring salts boron (B) and Se on soil microbial community composition associated with poplar trees is not known for bioremediation strategies. In this study, three Populus clones 13–366, 345–1, and 347–14 were grown in spring 2006 under highly saline, B, and Se clay-like soils in the west side of the San Joaquin Valley (SJV) of CA, as well as in non-saline sandy loam soils located in the east side of the SJV. After 7 years of growing in the respective soils of different qualities, soil samples were collected from poplar clones grown in saline and non-saline soils to examine and compare soil quality effects on soil microbial community biomass and composition. The phospholipid fatty acid (PLFA) analysis was used to characterize microbial community composition in soils from trees grown at both locations. This study showed that microbial biomass and the amount and proportion of arbuscular mycorrhizal fungal (AMF) community were lower in all three poplar clones grown in saline soil compared to non-saline soil. Amounts of Gram + bacterial and actinomycetes PLFAs were significantly lower in poplar clone 13–366 grown in saline soil compared to non-saline soil; however, they did not differ significantly in poplar clones 347–14 and 345–1. Additionally, amounts of saprophytic fungal, Gram ? bacterial and eukaryotic PLFA remained similar at saline and non-saline sites under poplar clones 347–14, 345–1, and 13–366. Therefore, this study suggested that salinity and B do have an impact on microbial biomass and AMF; however, these poplar clones still recycled sufficient amount of nutrients to support and protect saprophytic fungal and bacterial communities from the effects of poor quality soils.     

Radial and axial turgor pressure measurements in individual root cells of Mesembryanthemum crystallinum grown under various saline conditions

Abstract. Radial and axial turgor pressure profiles were measured with the pressure probe in untreated and salt-treated intact roots of Mesembryanthemum crystallinum. The microcapillary of the pressure probe was inserted step-wise into the root tissue 5, 25 and 50 mm away from the root cap. For evaluation of the data, only those recordings on a given root were used in which four discontinuous increases in turgor pressure occurred. These four turgor pressure increases could be related to the rhizodermal cells and to the cells in the three cortical layers. The measurements showed that a radial turgor pressure gradient of the same magnitude (directed from the third cortical layer to the external medium) existed along the root axis. The magnitude of this turgor pressure gradient decreased with increasing salinity (up to 400 mol m^-3 NaCl) in the growth medium. Addition of 10 mol m^-3 CaCl₂ to the 400 mol m^-3 NaCl medium partly reduced the salt-induced decrease in turgor pressure, but only in cells 25–50 mm away from the root tip. Combined with this effect, a small axial turgor pressure gradient was generated, therefore, in the cortex layers which was directed to the root tip. Measurements of the volumetric elastic modulus, ?, of the wall of the individual cells showed that the presence of salt considerably reduced the magnitude of this parameter and that addition of Ca²⁺ to the strongly saline medium partially diminished this decrease. This effect was strongest in cells 50 mm away from the root tip. The magnitude of ? of rhizodermal and cortical cells increased along the root axis both in untreated and in salt-treated roots. The ? value was significantly smaller for rhizodermal cells compared to the cortical cells, with the exception of cells 50 mm from the tip. In this tissue, rhizodermal and cortical cells exhibited nearly the same values. The decrease of the ?-values with salt and the increase along the root axis under the various growth conditions could be correlated with corresponding changes in cell volume. Diurnal changes in turgor pressure could not be detected in the individual root cells, with the notable exception of the rhizodermal and cortical cells located in the region 50 mm away from the root tip of the control plants. In these cells, an increase in turgor pressure was observed during the morning hours. Determination of the average osmotic pressure in tissue sections along the roots of control and salt-treated plants revealed that at 400 mol m^-3 NaCl the osmotic pressure gradient between the tissue and the medium is exo-directed, provided that the water is not (partly) immobilized.     

Carbon metabolism and bacteroid respiration in nodules of chick-pea (Cicer arietinum L.) plants grown under saline conditions

ABSTRACT

The present work investigates the relationships between nitrogen fixation, carbon metabolism and oxygen consumption by bacteroids of Mesorhizobium ciceri in root nodules of chick-pea plants. Its aim was to establish whether some of the compounds which accumulate under salt stress may be used as respiratory substrates by bacteroids to fuel their own metabolism and nitrogenase activity. Plants were grown in a growth chamber, and salt stress was induced by adding 50 mM NaCl to the nutrient solution at sowing. The data presented here show a rise in fermentative metabolism in nodules of chick-pea plants exposed to high salinity, and suggest that proline, lactate or ethanol, may play an important role as energy-yielding substrates for bacteroids in this plant species. The bacteroids could utilize glucose as a respiratory substrate both under control and saline conditions, while malate did not appear to be the preferred substrate in the presence of salt.     

Natural deuterium and oxygen-18 enrichment in leaf water of cotton plants grown under wet and dry conditions: evidence for water compartmentation and its dynamics

Abstract. Significant differences in leaf water oxygen and hydrogen isotopic composition were observed between cotton plants grown under wet and dry conditions. The magnitude of the differences could be fully explained by the conventional model that describes the isotopic composition of an evaporating water pool under steady state conditions. The results indicate that leaf water isotopic composition is strongly influenced by transpiration rate via its effects on relative humidity adjacent to the leaf surface and on the isotopic composition of the air moisture. Our application of the model, however, provides evidence that leaf water must consist of a mixture of several isotopically distinct pools. These pools are suggested to reside in the symplast, in the cell walls and intercellular spaces and in the veins. A model is proposed suggesting that only the water residing in the cell walls and the intercellular spaces (the transpiration pool) interacts directly with the external environment. The large symplastic pool responds to the external environment to a limited extent via its relatively slow exchange with water in the transpiration pool. It is likely that the isotopic composition of water in the symplastic pool is strongly buffered against shortterm environmental variations, a possibility that would have important implications for the isotopic conditions under which organic matter biosynthesis occurs.     

Hourly and seasonal variation in photosynthesis and stomatal conductance of soybean grown at future CO(2) and ozone concentrations for 3 years under fully open-air field conditions

It is anticipated that enrichment of the atmosphere with CO(2) will increase photosynthetic carbon assimilation in C3 plants. Analysis of controlled environment studies conducted to date indicates that plant growth at concentrations of carbon dioxide ([CO(2)]) anticipated for 2050 ( approximately 550 micromol mol(-1)) will stimulate leaf photosynthetic carbon assimilation (A) by 20 to 40%. Simultaneously, concentrations of tropospheric ozone ([O(3)]) are expected to increase by 2050, and growth in controlled environments at elevated [O(3)] significantly reduces A. However, the simultaneous effects of both increases on a major crop under open-air conditions have never been tested. Over three consecutive growing seasons > 4700 individual measurements of A, photosynthetic electron transport (J(PSII)) and stomatal conductance (g(s)) were measured on Glycine max (L.) Merr. (soybean). Experimental treatments used free-air gas concentration enrichment (FACE) technology in a fully replicated, factorial complete block design. The mean A in the control plots was 14.5 micromol m(-2) s(-1). At elevated [CO(2)], mean A was 24% higher and the treatment effect was statistically significant on 80% of days. There was a strong positive correlation between daytime maximum temperatures and mean daily integrated A at elevated [CO(2)], which accounted for much of the variation in CO(2) effect among days. The effect of elevated [CO(2)] on photosynthesis also tended to be greater under water stress conditions. The elevated [O(3)] treatment had no statistically significant effect on mean A, g(s) or J(PSII) on newly expanded leaves. Combined elevation of [CO(2)] and [O(3)] resulted in a slightly smaller increase in average A than when [CO(2)] alone was elevated, and was significantly greater than the control on 67% of days. Thus, the change in atmospheric composition predicted for the middle of this century will, based on the results of a 3 year open-air field experiment, have smaller effects on photosynthesis, g(s) and whole chain electron transport through photosystem II than predicted by the substantial literature on relevant controlled environment studies on soybean and likely most other C3 plants.     

Evidence of a direct role for Bcl-2 in the regulation of articular chondrocyte apoptosis under the conditions of serum withdrawal and retinoic acid treatment

The regulation of chondrocyte apoptosis in articular cartilage may underlay age-associated changes in cartilage and the development of osteoarthritis. Here we demonstrate the importance of Bcl-2 in regulating articular chondrocyte apoptosis in response to both serum withdrawal and retinoic acid treatment. Both stimuli induced apoptosis of primary human articular chondrocytes and a rat chondrocyte cell line as evidenced by the formation of DNA ladders. Apoptosis was accompanied by decreased expression of aggrecan, a chondrocyte specific matrix protein. The expression of Bcl-2 was downregulated by both agents based on Northern and Western analysis, while the level of Bax expression remained unchanged compared to control cells. The importance of Bcl-2 in regulating chondrocyte apoptosis was confirmed by creating cell lines overexpressing sense and antisense Bcl-2 mRNA. Multiple cell lines expressing antisense Bcl-2 displayed increased apoptosis even in the presence of 10% serum as compared to wild-type cells. In contrast, chondrocytes overexpressing Bcl-2 were resistant to apoptosis induced by both serum withdrawal and retinoic acid treatment. Finally, the expression of Bcl-2 did not block the decreased aggrecan expression in IRC cells treated with retinoic acid. We conclude that Bcl-2 plays an important role in the maintenance of articular chondrocyte survival and that retinoic acid inhibits aggrecan expression independent of the apoptotic process. J. Cell. Biochem. 71:302–309, 1998. © 1998 Wiley-Liss, Inc.     

Changes in CH4 and CO2 emissions from soils under flooded conditions after being exposed to FACE (free-air CO2 enrichment) for three years

To evaluate the variations of CO₂ and CH₄ emissions from FACE (free-air CO₂ enrichment, F) soils three years after rice-wheat rotation FACE treatment, incubation experiments in the laboratory with laboratory and elevated CO₂ concentration (1000 μl L?1) were carried out under flooded conditions at 25°C. Results show that soil organic carbon is increased by 11% after exposure to FACE treatment for three years. The results indicate that at laboratory and elevated CO₂, the cumulative CO₂ emissions from FACE soils are 35% and 22% higher than those from the ambient soils, whereas the cumulative CH₄ emissions from FACE soils are 2.6 and 2.3 times that of ambient soils. Thus, there is a larger ratio of cumulative emissions of CH₄ to CO₂ in the soil F. The elevated CO₂ concentration during the incubation stimulates the cumulative CO₂ emission significantly, but its stimulation on CH₄ emission is not statistically significant. The results indicate that the elevated atmospheric CO₂ concentration stimulates the turnover rates of soil organic matter, with a net increase in soil organic matter content, and alters the CH₄/CO₂ ratio.     

Changes in CH4 and CO2 emissions from soils under flooded conditions after being exposed to FACE (free-air CO2 enrichment) for three years

To evaluate the variations of CO₂ and CH₄ emissions from FACE (free-air CO₂ enrichment, F) soils three years after rice-wheat rotation FACE treatment, incubation experiments in the laboratory with laboratory and elevated CO₂ concentration (1000 μl L?1) were carried out under flooded conditions at 25°C. Results show that soil organic carbon is increased by 11% after exposure to FACE treatment for three years. The results indicate that at laboratory and elevated CO₂, the cumulative CO₂ emissions from FACE soils are 35% and 22% higher than those from the ambient soils, whereas the cumulative CH₄ emissions from FACE soils are 2.6 and 2.3 times that of ambient soils. Thus, there is a larger ratio of cumulative emissions of CH₄ to CO₂ in the soil F. The elevated CO₂ concentration during the incubation stimulates the cumulative CO₂ emission significantly, but its stimulation on CH₄ emission is not statistically significant. The results indicate that the elevated atmospheric CO₂ concentration stimulates the turnover rates of soil organic matter, with a net increase in soil organic matter content, and alters the CH₄/CO₂ ratio.