Water permeability of isolated cuticular membranes: The effect of cuticular waxes on diffusion of water

Summary The water permeability of astomatous cuticular membranes isolated from Citrus aurantium L. leaves, pear (Pyrus communis L.) leaves and onion (Allium cepa L.) bulb scales was determined before and after extraction of cuticular waxes with lipid solvents. In pear, the permeability coefficients for diffusion of tritiated water across cuticular membranes (CM) prior to extraction [P _d(CM)] decreased by a factor of four during leaf expansion. In all three species investigated P _d(CM) values of cuticular membranes from fully expanded leaves varied between 1 to 2×10^-7 cm^-3 s^-1·P _d(CM) values were not affected by pH. Extraction of cuticular waxes from the membranes increased their water permeability by a factor of 300 to 500. Permeability coefficients for diffusion of THO across the cutin matrix (MX) after extraction [P _d(MX)] increased with increasing pH. P _dvalues were not inversely proportional to the thickness of cuticular membranes. By treating the cutin matrix and cuticular waxes as two resistances acting in series it was shown that the water permeability of cuticles is completely determined by the waxes. The lack of the P _d(CM) values to respond to pH appeared to be due to structural effects of waxes in the cutin matrix. Cuticular membranes from the submerse leaves of the aquatic plant Potamogeton lucens L. were three orders of magnitude more permeable to water than the cuticular membranes of the terrestrial species investigated.Abbreviations CM cuticular membrane - MX cutin matrix - WAX waxes This study was supported by a grant from the Deutsche Forschungsgemeinschaft.     

Water permeability of plant cuticles: The effect of temperature on diffusion of water

The effect of temperature on water permeability of plant cuticles (astomatous Citrus leaf cuticles) has been investigated. The Arrhenius plot (logarithm of the permeability coefficient vs. 1/temperature) has two linear portions that intersect at 44° C. Evidence is presented to show that this intersection represents the solid/liquid phase transition of cuticular lipids. As the Arrhenius plot has only one phase transition in the temperature range of 5 to 80° C, it appears that all soluble cuticular lipids in the cuticle are present as a homogeneous mixture rather than as individual layers differing in composition. This view is supported by electron spin resonance evidence showing homogenous distribution of spin label fatty acids. The original distribution of soluble cuticular lipids is irreversibly altered by heating cuticular membranes above the transition temperature. This is accompanied by an irreversible increase in water peremeability, demonstrating the importance of the structure of cuticular lipids with regard to cuticular permeability.Abbreviations CM cuticular membranes - MX polymer matrix - SCL soluble cuticular lipids - MES morpholinoethane sulphonic acid - J flux - ESR electron spin resonance - THO tritiated water     

Water permeability of plant cuticles

Using the system vapor/membrane/liquid, permeability coefficients of cuticular transpiration (P _ct) were determined as functions of water activity in the vapor (a _wv). Enzymatically isolated cuticular membranes (CM) of Citrus aurantium L. and nonisolated CM of onion bulb scales and eggplant fruits were investigated. P _ct of Citrus and eggplant CM decreased with decreasing a _wv, while permeability coefficients of CM of onion were independent of a _wv. Extraction of soluble cuticular lipids (SCL) from the CM of Citrus increased permeability coefficients by a factor of approximately 500. This extraction had no effect on the dependence of P _ct on a _wv.Treating cuticular membranes as a resistance network consisting of SCL and the polymer matrix, it is shown that the permeability of onion CM is determined by the resistance of the SCL arranged in series with the polymer matrix. In this type of CM liquid and vapor are separated by a continuous, nonporous layer of SCL, and the driving force of transpiration is the gradient of partial pressure of water vapor across the SCL layer. In the CM of Citrus and eggplant, the SCL layer is traversed by polar pores that swell or shrink depending on a _wv. However, liquid continuity is maintained across these membranes down to a _wv=0.22, the lowest value used. In this type of membrane the driving force of transpiration is the water potential gradient across the membrane.Abbreviations CM cuticular membrane - MX polymer matrix - SCL soluble cuticular lipids - HEPES N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid - MES (N-morpholino)ethane sulfonic acid - SADH succinic acid 2,2-dimethyl hydrazide     

Ecophysiology of cuticular transpiration: comparative investigation of cuticular water permeability of plant species from different habitats

Water permeabilities of astomatous, isolated cuticular membranes (CM) of 24 different plants species were measured. Permeances varied from 1.7×10^–11 m·s^–1 (Vanilla planifolia leaf) up to 2.1×10^–9 m·s^–1 (Malus cf. domestica fruit) among different plant species, thus covering a range of over 2 orders of magnitude. Ranking of species according to permeances resulted in four distinct groups. The first group, of species with the lowest cuticular transpiration rates, included evergreen species growing in warm dry tropical climates (e.g. Vanilla planifolia and Monstera deliciosa leaves). The second class, with slightly higher water permeabilities, included evergreen species with typical scleromorphic leaf properties, adapted to a typical mediterranean type of climate with a dry period during the year (e.g. Citrus limon and Olea europaea leaves). The third group of species, where the highest leaf cuticular transpiration rates were observed, included deciduous species normally growing in a tempeate climate (e.g. Juglans regia and Forsythia suspensa leaves). Fruit cuticular membranes (CM) made up the fourth group (e.g. Capsicum annuum and Malus cf. domestica fruits), with even higher permeances than leaves of species from group 3. Thus, it appears that the plant species investigated show ecophysiological adaptations to the climatic demands of their natural habitats in cuticular water permeability.     

Gas permeability of plant cuticles

Cuticles from the adaxial surface of Citrus aurantium L. leaves and from the pericarp of Lycopersicon esculentum L. and Capsicum annuum L. were isolated enzymatically and their oxygen permeability was determined. Isolated cuticles were mounted between a gaseous and an aqueous compartment with the physiological outer side of the membrane facing the gaseous compartment. Permeability for oxygen was characterized by permeability (P) and diffusion (D) coefficients. P and D were independent of the driving force (gradient of oxygen concentration) across the cuticle, thus, Henry's law was obeyed. P values for the diffusion of oxygen varied between 3·10^-7 (Citrus), 1.4·10^-6 (Capsicum), and 1.1·10^-6 (Lycopersicon) m·s^-1. Extraction of soluble lipids from the cuticles increased the permeability. By treating the cutin matrix and the soluble lipids as resistances in series, it could be demonstrated that the soluble lipids were the main resistance for oxygen permeability in Citrus cuticles. However, in Lycopersicon and Capsicum, both the cutin matrix and the soluble lipids determined the total resistance. P values were not affected by either the proton concentration (pH 3–9) or the cations (Na⁺, Ca²⁺) present at the morphological inner side of the cuticles. It is concluded that the water content of cuticles does not affect the permeability properties for oxygen. Partition coefficients indicated a high solubility of oxygen in the cuticle of Citrus. The data suggest a solubility process in the cuticle of Citrus with respect to oxygen permeation.Abbreviations CM cuticular membrane - MX cutin polymer matrix - SCL soluble cuticular lipids     

Phase transitions and thermal expansion coefficients of plant cuticles

The temperature-induced volume expansion of enzymatically isolated cuticular membranes of twelve plant species was measured. All cuticular membranes exhibited distinct second-order phase transitions in the temperature range of about 40 to 50° C. Increases in the volumes of fruit cuticles (Lycopersicon, Cucumis, Capsicum, Solanum and Malus) were fully reversible, while leaf cuticular membranes (Ficus, Hedera, Nerium, Olea, Pyrus, Picea and Citrus) underwent irreversible structural changes. Below the phase-transition temperature, volumetric expansion coefficients ranged from 0.39·10^–6 m³·kg^–1·K^–1 to 0.62·10^–6 m³·kg^–1·K^–1, and above from 0.60·10⁶ m³·kg^–1·K^\-1 to 1.41· 10^–6 m³·kg^–1·K^–1. Densities of cuticles at 25° C ranged from 1020 kg·m^–3 to 1370 kg·m^–3. Expansion coefficients and phase transitions were characteristic properties of the polymer matrix as a composite material, rather than of cutin alone. It is argued that the sudden increase of water permeability above the transition temperature, is caused by an increase of disorder at the interface between the polymer matrix and the soluble cuticular lipids. Possible ecological and physiological consequences of these results for living plants are discussed.Abbreviations CM Cuticular membrane - CU cutin - MX polymer matrix - SCL soluble cuticular lipids (waxes) The authors greatfully acknowledge stimulating discussions with Drs. H. Gruler (Exp. Physik 3, Universität Ulm, FRG) and M. Riederer (Institut für Botanik und Mikrobiologie, Technische Universität München, München, FRG) and financial support by the Deutsche Forschungsgemeinschaft.     

The Use of the [13C]/[12C] Ratio for the Assay of the Microbial Oxidation of Hydrocarbons

The study deals with a comparative analysis of the relative abundances of the carbon isotopes ¹²C and ¹³C in the metabolites and biomass of the Burkholderia sp. BS3702 and Pseudomonas putida BS202-p strains capable of utilizing aliphatic (n-hexadecane) and aromatic (naphthalene) hydrocarbons as sources of carbon and energy. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of Burkholderia sp. BS3702 on n-hexadecane (¹³C = –44.6 ± 0.2) were characterized by the values of ¹³C_{CO 2} = –50.2 ± 0.4, ¹³C_biom = –46.6 ± 0.4, and ¹³C_exo = –41.5 ± 0.4, respectively. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of the same bacterial strain on naphthalene (¹³C = –21 ± 0.4) were characterized by the isotope effects ¹³C_{CO 2} = –24.1 ± 0.4, ¹³C_biom = –19.2 ± 0.4, and ¹³C_exo = –19.1 ± 0.4, respectively. The possibility of using the isotope composition of metabolic carbon dioxide for the rapid monitoring of the microbial degradation of petroleum hydrocarbons in the environment is discussed.     

Transport properties of cuticular waxes of Fagus sylvatica L. and Picea abies (L.) Karst.: Estimation of size selectivity and tortuosity from diffusion coefficients of aliphatic molecules

Diffusion coefficients for thirteen lipophilic organic compounds in reconstituted waxes of Fagus sylvatica L. and Picea abies (L.) Karst. were measured to characterise the transport properties of the cuticular waxes that form the transport-limiting barrier of plant cuticles. Desorption kinetics (relative amounts desorbed versus time) were asymptotic, but could be linearized up to 50% desorption by plotting relative amounts desorbed versus the square root of time. Diffusion coefficients calculated from the slopes of the linear regressions ranged from 10^–22 to 10^–17 m²·s^–1 and decreased with increasing molecular size. This size dependency of diffusion coefficients was analysed, assuming an exponential dependence of the diffusion coefficients on molar volumes, which allowed cuticular transport properties to be related to the physical structure of the wax. Furthermore, the fact that the barrier properties of Fagus wax are less pronounced than those of Picea is interpreted as an ecological adaptation of the respective tree species to their habitats at the level of their cuticular transport barriers.Abbreviation D diffusion coefficient The authors gratefully acknowledge financial support by the Schwerpunkt Programm Baumphysiologie of the Deutsche Forschungsgemeinschaft.     

The effect of the environment on the permeability and composition of Citrus leaf cuticles

The constituents of the soluble cuticular lipids (SCL) of the leaf blades of Citrus aurantium L. were identified by gas chromatography-mass spectrometry and quantified. Major components were 1-alkanols (C₂₄ to C₄₀), n-alkyl esters (C₃₆ to C₅₆), n-alkanoic acids (C₂₈ to C₃₄), n-alkanes (C₂₂ to C₄₀) and triterpenones, while n-alkanals (C₂₉ to C₃₈), sterols, and alkyl benzenes (molecular weights 260, 274 and 288) made minor contributions. Leaf age and side significantly affected the quantitative composition of SCL. Increased day temperature during the development of leaves led to decreased amounts per unit area of n-alkanes, 1-alkanols, n-alkanoic acids and n-alkyl esters while increased night temperatures resulted in increased amounts of n-alkanes n-alkanoic acids and 1-alkanols. Relative humidity had no effect on the amounts or composition of SCL. The permeability of cuticular membranes to water (described in part I of this paper) and the composition of SCL were not related. A model for the molecular structure of the transport-limiting barrier of plant cuticles and for the transport of water across it is proposed.Abbreviations CM cuticular membrane - GC gas chromatogra-phy - MS mass spectroscopy - TLC thin-layer (planar) chromatography - SCL soluble cuticular lipids The authors are indebted to Dr. R. Winkler and H. Krause, Laboratorium für Strukturchemie des Fachbereichs Chemie, Biologie und Geowissenschaften, Technische Universität München, FRG, for performing the GC-MS analyses and their valuable help in the identification of SCL constituents. This work has been supported by the Deutsche Forschungsgemeinschaft and the Bayerische Staatsministerium für Wissenschaft und Kunst.     

n-3 PUFA productivity in chemostat cultures of microalgae

Eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid productivities from chemostat cultures of an isolate of Isochrysis galbana have been studied. The productivities reached in the interval of dilution rates between 0.0295 h^–1 and 0.0355 h^–1 were 1.5mg·1^–1·h^–1 for lipids, 300 g·1^–1·h^–1 for EPA and 130g1·1^–1·h^–1 for DHA. Furthermore, light attenuation by mutual shading, and agitation speed influences on growth and fatty acid composition were analysed. A model relating steady-state dilution rates to internal average light intensity has been proposed, the parameter values of which obtained by non-linear regression were: maximum specific growth rate (_max)=0.0426 h^–1; the affinity of cells to light (I_k) = 10.92 W·m^–2; the exponent (n) = 5.13; regression coefficient (r ²)=0.9999. Correspondence to: E. Molina Grima     

Ion exchange properties of isolated tomato fruit cuticular membrane: Exchange capacity,nature of fixed charges and cation selectivity

Summary Isolated tomato fruit cuticular membrane, free of extractable materials, was titrated potentiometrically using various bases. Three dissociable groups were observed in the pH ranges 3–6 (0.2 meq g^-1), 6–9 (0.3 meq g^-1) and 9–12 (0.55 meq g^-1). The first group was tentatively assigned to-COOH groups of pectic materials and protein embedded in the membrane, the second to nonesterified-COOH groups of the cutin polymer and the third to phenolic-OH groups, such as non-extractable flavenoids present in the membrane, and to a small amount of-NH ₃ ⁺ groups of proteins. The cuticular membrane exhibited a behavior typical of highly cross-linked, high-capacity ion exchange resins of the weak-acid type. Ion exchange capacity increased with increasing pH and neutral salt concentration. At constant pH and salt concentration, the exchange capacity increased with increasing counter ion valence and decreasing crystal radius, e.g. [tris (ethylenediamine) Co]³⁺Ca²⁺>Ba²⁺>Li⁺>Na⁺>Rb⁺>N(CH₃) ₄ ⁺ . The cutin polymer exhibited a pronounced selectivity for Ca²⁺ over Na⁺ which increased with increasing neutralization of fixed charges. The large trivalent [Co(en)₃]³⁺ was preferred only at low equivalent ionic fractions in the polymer. These results are discussed in relation to the structure and function of cuticular membranes.     

The electrical response ofAvena coleoptile cortex to auxins

Solute mobilities of 28 compounds in isolated cuticular membranes (CM) from Capsicum annuum L. fruit, Citrus aurantium L. and Pyrus communis L. leaves were studied using unilateral desorption from the outer surface. First-order rate constants of desorption (k^*), which are directly proportional to the diffusion coefficient in the waxy outer limiting skins of cuticles were measured. When log k^* was plotted vs. molar volumes of test compounds linear graphs were obtained. The y-intercepts of these graphs (k^*) represent the mobility of a hypothetical molecule having zero molar volume and the slopes of the graphs () represent the size selectivity of the barrier and are related to the free volume available for diffusion. Thus, solute mobilities in cuticles are composed of two independent terms which are subtractive. If k^* and are known, k^* can be estimated for any solute from its molar volume (V_x) using the equation log k^*=log k^* –V_x. These parameters were used to analyse the effects of plant species, extraction of cuticular waxes and molecular structure of solutes on solute mobilities in plant cuticles. For aliphatic solutes, k^* was a factor of 10 smaller than for cyclic compounds, while was 0.011 and 0.012, respectively. The k^*-values for CM of the three species were very similar, but was higher for bitter-orange CM (0.012) than for those of pepper fruits and pear leaves (0.009). This has the consequence that differences in solute mobilities (k^*) among cuticles from different plan species increase with increasing molar volumes of solutes. Our data and our analysis provide evidence that constituents of cuticular waxes are mobile, at least in the solid amorphous wax fraction, but mobility decreases rapidly with increasing molar volume. For instance, if amounts to 0.01, mobilities of wax monomers decrease by a factor of 10 for every increase in molar volume of 100 cm³ · mol^–1. Thus, hexadecanoic acid is quite mobile in the amorphous wax fraction of Citrus (k^*=1.5×10^–6·s^–1), but for dotriacontane having twice the molar volume, k^* was only 2.5×10^–9·s^–1, which is almost three orders of magnitude smaller. Wax esters have even higher molar volumes and their mobilities will be even smaller (about 4×10^–12·s^–1 for a C₄₈-ester). Since low chain mobilities are a prerequisite for low mobilities and permeabilities, the selective advantage of high-molecular-weight wax monomers in plant cuticular waxes becomes obvious. Extracting cuticular waxes from pear leaf CM increased solute mobilities by a factor of 182, but it had no effect on size selectivity. We interpret this result as evidence to the effect that cuticular waxes reduce mobility by increasing tortuosity of the diffusion path, rather than by decreasing the mean free path of diffusional jumps and jump frequencies of diffusants.Abbreviations CM cuticular membrane(s) - 2,4-D 2,4-dichloro-phenoxyacetic acid - LAB lactic acid buffer - MX polymer matrix membranes - UDOS unilateral desorption from the outer surface     

Wax and suberin development of native and wound periderm of potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) and its relation to peridermal transpiration

Native and wound periderm was isolated enzymatically from potato (Solanum tuberosum L. cv. Desirée) tubers at different time intervals between 0 days up to 4 weeks after harvesting. Wound periderm formation was induced by carefully removing native periderm from freshly harvested tubers before storage. The chemical composition of lipids (waxes) obtained by chloroform extraction, as well as the monomeric composition of native and wound suberin polymer after transesterification by boron trifluoride/methanol, was analyzed using gas chromatography and mass spectrometry. Both types of periderm contained up to 20% extractable lipids. Besides linear long-chain aliphatic wax compounds, alkyl ferulates were detected as significant constituents. In wound periderm they amounted to more than 60% of the total extracts. Within 1 month of storage, suberin amounts in the polymer increased 2-fold in native periderm (180 g cm^–2), whereas in wound periderm about 75.0 g cm^–2 suberin polymer was newly synthesized. Native potato tuber periderm developed a very efficient transport barrier for water with permeances decreasing from 6.4×10^–10 m s^–1 to 5.5×10^–11 m s^–1 within 1 month of storage. However, the water permeability of wound periderm was on average 100 times higher with permeances decreasing from 4.7×10^–8 m s^–1 after 3 days to only 5.4×10^–9 m s^–1 after 1 month of storage, although suberin and wax amounts in wound periderm amounted to about 60% of native periderm. From this result it must be concluded that the occurrence of suberin with wax depositions in cell walls does not necessarily allow us to conclude that these cell walls must be nearly perfect barriers to water transport. In addition to the occurrence of the lipophilic biopolymer suberin and associated waxes, the still unknown molecular arrangement and precisely localized deposition of suberin within the cell wall must contribute to the efficiency of suberin as a barrier to water transport.     

Permeation characteristics of isolated cuticles of Citrus aurantium L. for monoterpenes

Summary Permeation parameters of isolated cuticular membranes of Citrus aurantium L. for gaseous monoterpenes were determined by an isostatic system. For -pinene and d-limonene permeability coefficients range from 4.3 × 10^–11 m^–2 s^–1 to 7.3 × 10^–11 m^–2 s^–1. These values can be compared to that measured for benzene gas at the cuticle of Citrus. The permeability coefficients of the two monoterpenes did not differ significantly, in contrast to their diffusioin coefficients. The diffusion coefficient values are 3.7 × 10^–15 m^–2 s^–1 for limonene and 15.5 × 10^–15 m^–2 s^–1 for -pinene. The reason for this difference is still unclear. A dependence of the permeation parameters on the direction of the monoterpene transport could not be observed. Moreover, there are some indications that, in spite of its heterogeneous character, the cuticular membrane of Citrus is homogeneous in respect to the transport of small gaseous molecules. An exposure to environmentally relevant ozone concentrations for 6 months did not change the permeation characteristics of the membrane. Due to the high variability of the samples only a tendency towards higher permeability coefficients of cuticles treated with 80 ppb ozone was observed. This may be attributed to a reduced tension of the membrane caused by chain fractions.This paper is dedicated to Prof. Dr. Otto Härtl, Graz, on the occasion of his 80th birthday.     

Proton conductance through phospholipid bilayers: Water wires or weak acids?

The proton/hydroxide (H⁺/OH^–) permeability of phospholipid bilayer membranes at neutral pH is at least five orders of magnitude higher than the alkali or halide ion permeability, but the mechanism(s) of H⁺/OH^– transport are unknown. This review describes the characteristics of H⁺/OH^– permeability and conductance through several types of planar phospholipid bilayer membranes. At pH7, the H⁺/OH^– conductances (G _H/OH) range from 2–6 nS cm^–2, corresponding to net H⁺/OH^– permeabilities of (0.4–1.7)×10^–5 cm sec^–1. Inhibitors ofG _H/OH include serum albumin, phloretin, glycerol, and low pH. Enhancers ofG _H/OH include chlorodecane, fatty acids, gramicidin, and voltages >80 mV. Water permeability andG _H/OH are not correlated. The characteristics ofG _H/OH in fatty acid (weak acid) containing membranes are qualitatively similar to the controls in at least eight different respects. The characteristics ofG _H/OH in gramicidin (water wire) containing membranes are qualitatively different from the controls in at least four different respects. Thus, the simplest explanation for the data is thatG _H/OH in unmodified bilayers is due primarily to weakly acidic contaminants which act as proton carriers at physiological pH. However, at low pH or in the presence of inhibitors, a residualG _H/OH remains which may be due to water wires, hydrated defects, or other mechanisms.     

Permeability of ammonia,methylamine and ethylamine in the cyanobacterium,Synechococcus R-2 (Anacystis nidulans) PCC 7942

Summary Permeabilities of ammonia (NH₃), methylamine (CH₃NH₂) and ethylamine (CH₃CH₂NH₂) in the cyanobacterium (cyanophyte)Synechococcus R-2 (Anacystis nidulans) have been measured. Based on net uptake rates of DCMU (dichlorophenyldimethylurea) treated cells, the permeability of ammonia was 6.44±1.22 m sec^–1 (n=13). The permeabilities of methylamine and ethylamine, based on steady-state¹⁴C labeling were more than ten times that of ammonia (P _methylamine=84.6±9.47 m sec^–1 (76),P _ethylamine=109±11 m sec^–1 (55)). The apparent permeabilities based on net uptake rates of methylamine and ethylamine uptake were significantly lower, but this effect was partially reversible by ammonia, suggesting that net amine fluxes are rate limited by proton fluxes to an upper limit of about 700 nmol m^–2 sec^–1. Increasing concentrations of amines in alkaline conditions partially dissipated the pH gradient across the cell membrane, and this property could be used to calculate the relative permeabilities of different amines. The ratio of ethylamine to methylamine permeabilities was not significantly different from that calculated from the direct measurements of permeabilities; ammonia was much less effective in dissipating the pH gradient across the cell membrane than methylamine or ethylamine. An apparent permeability of ammonia of 5.7±0.9 m sec^–1 could be calculated from the permeability ratio of ammonia to methylamine and the experimentally measured permeability of methylamine. The permeability properties of ammonia and methylamine are very different; this poses problems in the interpretation of experiments where¹⁴C-methylamine is used as an ammonia analogue.     

Plant response to drought stress simulated by ABA application: Changes in chemical composition of cuticular waxes

Plant cuticles form the interface between epidermal plant cells and the atmosphere. The cuticle creates an effective barrier against water loss, bacterial and fungal infection and also protects plant tissue from UV radiation. It is composed of the cutin matrix and embedded soluble lipids also called waxes. Chemical composition of cuticular waxes and physiological properties of cuticles are affected by internal regulatory mechanisms and environmental conditions (e.g. drought, light, and humidity). Here, we tested the effect of drought stress simulation by the exogenous application of abscisic acid (ABA) on cuticular wax amount and composition. ABA-treated plants and control plants differed in total aboveground biomass, leaf area, stomatal density and aperture, and carbon isotope composition. They did not differ in total wax amount per area but there were peculiar differences in the abundance of particular components. ABA-treated plants contained significantly higher proportions of aliphatic components characterized by chain length larger than C₂₆, compared to control plants. This trend was consistent both between and within different functional groups of wax components. This can lead to a higher hydrophobicity of the cuticular transpiration barrier and thus decrease cuticular water loss in ABA-treated plants. At both ABA-treated and control plants alcohols with chain length C₂₄ and C₂₆ were predominant. Such a shift towards wax compounds having a higher average chain length under drought conditions can be interpreted as an adaptive response of plants towards drought stress.     

Homologous long-chain δ-lactones in leaf cuticular waxes of Cerinthe minor

A homologous series of eleven δ-lactones (1,5-alkanolides) was identified in cuticular waxes from leaves of Cerinthe minor L., six of them representing novel compounds. They accounted for 79% of the total coverage of 41 μg wax per cm² leaf area. Various chemical transformations with product identification by GC-mass spectrometry and GC-FTIR were employed to assign the structures. The chain-lengths of the δ-lactones ranged from C₂₂ to C₃₂ and even-numbered homologues were prevalent. Additionally, aldehydes (C₂₆–C₃₀), alkanes (C₂₃–C₂₉), primary alcohols (C₂₆–C₃₂), alkanoic acids (C₂₀–C₃₂), wax esters (C₄₀–C₅₆) and lupeol were detected.     

Enteromorpha as a monitor of heavy metals in estuaries

An account is given of the use of Enteromorpha to monitor zinc, cadmium, mercury and lead pollution in six estuaries and the British North Sea coast. The ranges for each element were: Zn, 19–437 µg g^–1; µg g^–1 Cd, 0.07–4.8 µg g^–1; Hg, 0.02–0.23 µg g^–1. It is suggested that tissue analysis of Enteromorpha is one of the most useful biological techniques available in estuaries for pin-pointing aqueous (as opposed to sediment) metal contamination, and also for providing data suitable for world-wide comparisons. Provisional values are given for concentrations corresponding to moderate and high pollution.Deceased     

Water permeability and lipid composition of toad urinary bladder: The influence of temperature

Summary The water diffusional permeability, its activation energy and the lipid composition were studied in urinary bladders from toads adapted to different temperatures. It was observed that the unidirectional water flux greatly depends on the temperature at which the experiments are performed. This dependence is greater in the animals adapted to higher temperatures. Toads adapted to cold show strong reduction in the activation energy for water diffusion permeability (from 11.4±1.9 kcal·mol^–1 to 4.4±1.1 kcal·mol^–1) and an increase of 30% in the amount of total lipids from bladder epithelial cells. There were no significant changes in the phospholipid/cholesterol ratio, composition of the paraffinic chains or protein concentration between toads adapted to both temperatures. The possibility that water translocates through the mucosal border of the toad bladder by partitioning in the polar zone and diffusioning between the hydrocarbon chains of the membrane lipids and that cold adaptation would induce a stronger packing of lipids in the membrane is discussed.