COMPARISON OF METHYL JASMONATE AND CADMIUM EFFECT ON SELECTED PHYSIOLOGICAL PARAMETERS IN SCENEDESMUS QUADRICAUDA (CHLOROPHYTA,CHLOROPHYCEAE)1

Effect of methyl jasmonate (MeJA) in the concentrations of 10 or 100 μM on selected physiological parameters in Scenedesmus quadricauda (Turp.) Bréb. after 24 h of exposure was studied. Results were compared with the application of general toxic metal (cadmium, Cd) to identify MeJA‐specific responses. Accumulation of reactive oxygen species (ROS; hydrogen peroxide and superoxide) was the most elevated by 10 μM MeJA and 100 μM Cd, while total chls showed decrease (Cd) and increase (MeJA) in these variants. The amount of carotenoids and cell viability were affected neither by MeJA nor by Cd application. The sum of free amino acids was considerably elevated by 10 μM Cd (increase in histidine, threonine, arginine, leucine, and lysine mainly) but depleted by 100 μM MeJA (14 from 17 compounds decreased), while accumulation of soluble proteins was unaffected by Cd and enhanced by MeJA. Cadmium application reduced the amount of Ca and also Mg in the case of 100 μM Cd, while MeJA had no effect on the content of mineral nutrients. Total Cd content reached 557 and 1,334 μg · g^?1 dry weight (dwt) in 10 and 100 μM Cd variant, respectively. Intracellular Cd uptake was ca. 55% from total Cd content in both Cd variants. The present findings are discussed in the context of the available literature, and possible explanations are suggested.     

菠萝叶片中有机酸含量、转氨酶和脱氢酶活性的昼夜变化

研究了菠萝叶片中苹果酸、柠檬酸、草酰乙酸、α－酮戊二酸和草酸等有机酸和转氨酶、脱氢酶活性的昼夜变化。结果表明,除了草酸外,其他４种有机酸的含量都呈白天降低晚上增高的趋势。有机酸的含量在上午９∶００时有最高值,下午１８∶００时有最低值。苹果酸、柠檬酸、草酰乙酸、α－酮戊二酸和草酸的昼夜最大差值分别为１４０、６１、３１．８、８．２和６．７ｍｍｏｌ·ｋｇ－１ＦＷ。异柠檬酸脱氢酶和琥珀酸脱氢酶活性的昼夜变化与有机酸的趋势一致,白天降低晚上增高。可溶性蛋白和游离氨基酸含量以及谷草转氨酸、谷丙转氨酶活性在下午１８∶００时有最高值,凝胶电泳显示菠萝叶片中有新的蛋白带出现。     

FATTY ACIDS AND HYDROXY FATTY ACIDS IN THREE SPECIES OF FRESHWATER EUSTIGMATOPHYTES

The monocarboxylic fatty acids and hydroxy fatty acids of three species of freshwater microalgae—Vischeria punctata Vischer, Vischeria helvetica (Vischer et Pascher) Taylor, and Eustigmatos vischeri (Hulbert) Taylor, all from the class Eustigmatophyceae— were examined. Each species displayed a very similar distribution of fatty acids, the most abundant of which were 20:5n-3, 16:0, and 16:1n-7; C₁₈ polyunsaturated fatty acids were minor components. These fatty acid distributions closely resemble those found in marine eustigmatophytes but are quite distinct from those found in most other algal classes. These microalgae also contain long-chain saturated and unsaturated monohydroxy fatty acids. Two distinct types of hydroxy fatty acids were found: a series of saturated α-hydroxy acids ranging from C₂₄ to C₃₀ with a shorter series of monounsaturated α-hydroxy acids ranging from C₂₆ to C₃₀ together with a series of saturated β-hydroxy acids ranging from C₂₆ to C₃₀. The latter have not previously been reported in either marine or freshwater microalgae, although C₃₀ to C₃₄ midchain (ω-18)-hydroxy fatty acids have been identified in hydrolyzed extracts from marine eustigmatophytes of the genus Nannochloropsis, and C₂₂ to C₂₆ saturated and monounsaturated α-hydroxy fatty acids have been found in three marine chlorophytes. These findings have provided a more complete picture of the lipid distributions within this little studied group of microalgae as well as a range of unusual compounds that might prove useful chemotaxonomic markers. The functions of the hydroxy fatty acids are not known, but a link to the formation of the lipid precursors of highly aliphatic biopolymers is suggested.     

CHANGES IN PHOTOSYNTHETIC YIELD,AMINO ACIDS,AND ORGANIC ACIDS ARE INDUCED BY AMMONIUM ADDITION TO CELLS OF PHORMIDIUM LAMINOSUM (CYANOPHYCEAE)1

The effect of NH₄⁺ addition to NO₃^?-growing cells of the non-N₂-fixing cyanobacterium Phormidium laminosum (Agardh) Gomont (strain OH-1-pCl₁) on photo-synthetic and respiratory electron transport as well as on the intracellular levels of amino acids and some organic acids was studied. Addition of ammonium to nitrate-growing cells resulted in substantial increases in the pool size of most amino acids and a transient decrease in the pool size of organic acids. The high demand for organic acids was partially overcome by degradation of stored carbohydrates, more than by newly fixed carbon, as indicated by the large stimulation of the respiration rate upon ammonium addition. Following ammonium addition, the photosynthetic yield of the in vivo noncyclic electron transport decreased, and the sensitivity of photosystem II to photodamage increased. Results indicate that cells balance their photosynthetic and respiratory activities depending on nitrogen availability and point to an important involvement of respiration in providing energy for ammonium assimilation until adaptation of bioenergetic processes to the new nitrogen source is complete.     

THE KINETICS OF ORGANIC ACID UPTAKE BY THREE CHLOROPHYTA IN AXENIC CULTURE1

The uptake of 8 ¹⁴C-labelled dissolved carboxylic acids by Ankistrodesmus falcatus (Corda) Ralfs, Chlamydomonas segnis Ettl and Chlorella sp. was measured. The acids were acetic, citric, fumaric, glycollic, lactic, malic, pyruvic and succinic acids. For some substrates uptake was active, but for others it was apparently passive. The respiratory loss of substrates ranged from 5.0 to 95.0% of the total amount of C assimilated. The potential ability of these organisms to compete with bacterial populations for such substrates in natural waters was examined. In terms of the kinetics of assimilation it appeared that A. falcatus might compete successfully for glycollate, lactate, pyruvate and succinate; C. segnis for glycollate and lactate and Chlorella sp. for lactate, malate and possibly glycollate. Calculated replacement times of the algae on the 8 substrates did, however, indicate that although they might compete successfully for some substrates they were generally unable to grow upon them at the concentrations provided.     

PHYLOGENETIC VARIATION IN THE TOLERANCE AND UPTAKE OF ORGANIC CONTAMINANTS

An investigation into the phylogenetic variation of plant tolerance and the root and shoot uptake of organic contaminants was undertaken. The aim was to determine if particular families or genera were tolerant of, or accumulated organic pollutants. Data were collected from sixty-nine studies. The variation between experiments was accounted for using a residual maximum likelihood analysis to approximate means for individual taxa. A nested ANOVA was subsequently used to determine differences at a number of differing phylogenetic levels. Significant differences were observed at a number of phylogenetic levels for the tolerance to TPH, the root concentration factor and the shoot concentration factor. There was no correlation between the uptake of organic pollutants and that of heavy metals. The data indicate that plant phylogeny is an important influence on both the plant tolerance and uptake of organic pollutants. If this study can be expanded, such information can be used when designing plantings for phytoremediation or risk reduction during the restoration of contaminated sites.     

金鱼对铅和镉的吸收蓄积

研究了铅、镉暴露实验中金鱼不同器官经有收积累铅、镉的过程及蓄积量。结合到金鱼鳃上的铅、镉可用Langmuir吸附等温式描述。最大结合容量Bmax为：铅0.9424mmol/kg干重;镉17662μmol/kg条件稳定常数logK为：铅7.80;镉6.38。Hill结合特征常数为：铅1.012;镉1.175,表明金鱼鳃上仅有一种类型的Pb^2 或Cd^2 结合位点。鳃中铅、镉含量与血液含量以及血液铅、镉含量与肾、肝中的含量呈正相关关系。肾脏对铅、镉的蓄积量高于其他器官。     

SCANNING ELECTRON MICROSCOPY OBSERVATIONS OF DEFORMITIES IN SMALL PENNATE DIATOMS EXPOSED TO HIGH CADMIUM CONCENTRATIONS1

Different types of malformations are likely to affect the morphology of diatoms when exposed to particularly unstable environmental conditions, the most easily identifiable being distortion of the whole frustule. In the present study, we investigated, by means of SEM, valve abnormalities induced by high cadmium contamination (100 μg · L^?1) in small pennate diatoms. Changes in the shape of Amphora pediculus (Kütz.) Grunow and anomalous sculpturing of the cell wall of many species, such as Encyonema minutum (Hilse) D. G. Mann, Mayamaea agrestris (Hust.) Lange‐Bert., Gomphonema parvulum (Kütz.) Kütz., or Eolimna minima (Grunow) Lange‐Bert., were observed, which were not, or almost not, noticeable in the LM. With consideration to current knowledge of diatom morphogenesis, metal uptake by the cell would induce, directly or indirectly, damage to many cytoplasmic components (e.g., microtubules, cytoskeleton, Golgi‐derived vesicles) involved in the precisely organized silica deposition. This study confirms that many species, whatever their size, are likely to exhibit morphological abnormalities under cadmium stress, and that this indicator may be valuable for the biomonitoring of metal contamination, even if SEM observations are not necessary for routine studies.     

CADMIUM AND MANGANESE ACCUMULATION IN PHYTOLACCA AMERICANA L. AND THE ROLES OF NON-PROTEIN THIOLS AND ORGANIC ACIDS

Phytolacca americana L. can accumulate large amounts of heavy metals in its aerial tissues, especially cadmium (Cd) and manganese (Mn). It has great potential for use in phytoextraction of metals from multi-metal-contaminated soils. This study was conducted to further investigate the Cd- and Mn-tolerance strategies of this plant. Concentrations of non-protein thiols (NPTs) and phytochelatins (PCs) in leaves and roots increased significantly as the concentration of Cd in solution increased. The molar ratios of PCs:soluble Cd ranged from 1.8 to 3.6 in roots and 8.1 to 31.6 in leaves, suggesting that the cellular response involving PC synthesis was sufficient to complex Cd ions in the cytosol, especially that of leaves. In contrast, excess Mn treatments did not result in a significant increase in NPT or PC concentrations in leaves or roots. Oxalic acid concentrations in leaves of plants exposed to 2 or 20 mM Mn reached 69.4 to 89.3 mg (0.771 to 0.992 mmol) g^–1 dry weight, respectively, which was approximately 3.7- to 8.6-fold higher than the Mn level in the 0.6 M HCl extract. Thus, oxalic acid may play an important role in the detoxification of Mn.     

UPTAKE OF LIPOPHILIC CADMIUM COMPLEXES BY THREE GREEN ALGAE: INFLUENCE OF HUMIC ACID AND ITS pH DEPENDENCE1

Cadmium forms neutral, lipophilic CdL₂⁰ complexes with diethyldithiocarbamate (L = DDC) and with ethylxanthate (L = XANT). In a synthetic solution and in the absence of natural dissolved organic matter (DOM), for a given total Cd concentration, uptake of these complexes by unicellular algae is much faster than the uptake of the free Cd²⁺ cation. The objective of the present study was to determine how this enhanced uptake of the lipophilic CdL₂⁰ complexes was affected by the presence of natural DOM (Suwannee River humic acid, SRHA). Experiments were performed with Cd(DDC)₂⁰ and Cd(XANT)₂⁰ at two pH values (7.0 and 5.5) and with the three chlorophytes [Chlamydomonas reinhardtii P. A. Dang., Pseudokirchneriella subcapitata (Korshikov) Hindák, Chlorella fusca var. vacuolata Shihira et R. W. Krauss]. Short‐term uptake (30–40 min) of the CdL₂⁰ complexes was followed in the absence and presence of SRHA (6.5 mg C · L^?1). Acidification from pH 7.0 to 5.5 decreased CdL₂⁰ uptake by the three algae, in the presence or absence of humic acid (HA). The dominant effect of the HA was to decrease Cd uptake, due to its interaction with the CdL₂⁰ complexes in solution. However, if uptake of the free CdL₂⁰ complexes was compared in the presence and absence of HA, in four of eight cases initial uptake rate constants (k_i) were significantly higher (P < 0.05) in the presence of the HA, suggesting the operation of an interfacial effect of the HA at the algal cell membrane, favoring uptake of CdL₂⁰. Overall, the experimental results suggest that neutral metal complexes will be less bioavailable in natural waters than they are in synthetic laboratory media in the absence of natural DOM.     

VETIVER GRASS,VETIVERIA ZIZANIOIDES: A CHOICE PLANT FOR PHYTOREMEDIATION OF HEAVY METALS AND ORGANIC WASTES

Glasshouse and field studies showed that Vetiver grass can produce high biomass (>100t/tha^?1 year^?1) and highly tolerate extreme climatic variation such as prolonged drought, flood, submergence and temperatures (?15°–55°C), soils high in acidity and alkalinity (pH 3.3–9.5), high levels of Al (85% saturation percentage), Mn (578 mg kg^?1), soil salinity (ECse 47.5 dS m^?1), sodicity (ESP 48%), and a wide range of heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb, Se, and Zn). Vetiver can accumulate heavy metals, particularly lead (shoot 0.4% and root 1%) and zinc (shoot and root 1%). The majority of heavy metals are accumulated in roots thus suitable for phytostabilization, and for phytoextraction with addition of chelating agents. Vetiver can also absorb and promote biodegradation of organic wastes (2,4,6-trinitroluene, phenol, ethidium bromide, benzo[a]pyrene, atrazine). Although Vetiver is not as effective as some other species in heavy metal accumulation, very few plants in the literature have a wide range of tolerance to extremely adverse conditions of climate and growing medium (soil, sand, and tailings) combined into one plant as vetiver. All these special characteristics make vetiver a choice plant for phytoremediation of heavy metals and organic wastes.     

萘乙酸、激动素和伤害对绿豆子叶愈伤组织形成的作用以及和色氨酸、吲(口朶)乙酸生物合成的关系

本文以绿豆子叶为材料研究了切伤、外源萘乙酸及激动素诱导形成愈伤组织的作用及其与内源色氨酸和吲哚乙酸生物合成的关系。实验结果表明,切伤对于愈伤组织的形成具有重要作用,切伤面积的大小与愈伤组织的增殖成正比。在绿豆子叶愈伤组织形成的初期,游离色氨酸和内源吲哚乙酸的水平均降低,而在后期,组织内部游离色氨酸和吲哚乙酸的含量都有增加。在培养基中加入外源色氨酸可以部分代替萘乙酸促进愈伤组织的形成。可以认为,外源激素诱导愈伤组织的形成是通过促进内源色氨酸和内源吲哚乙酸的生物合成而实现的。受伤对愈伤组织的形成也起了重要的协同作用。     

乙酸积累对基因工程菌培养的影响及与培养基pH的关系

在rIL-2工程菌K_(802)(pLY—4)高密度培养中,发现培养液中有大量代谢副产物乙酸积累,乙酸的存在对工程菌的生长和产物的表达均有明显的抑制作用,这种抑制作用是制约工程菌高密度培养的重要因素。为了减小这种抑制作用,初步研究了培养基pH与乙酸抑制作用的关系,发现适当提高培养基pH值,能减小乙酸的抑制作用;高密度培养时,提高培养基的pH后,虽然仍有大量乙酸积累,但产物的表达水平和菌密度都有提高。     

SYNTHESIS OF MYCOSPORINE-LIKE AMINO ACIDS IN CHONDRUS CRISPUS (FLORIDEOPHYCEAE) AND THE CONSEQUENCES FOR SENSITIVITY TO ULTRAVIOLET B RADIATION

The induction and protective role of the UV-absorbing compounds known as mycosporine-like amino acids (MAAs) were examined in sublittoral Chondrus crispus Stackh. transplanted for 2 weeks in the spring and summer to shallow water under three irradiance conditions: PAR (photosynthetically active radiation; 400–700 nm), PAR + UVA (PAR + 320– 400 nm), PAR + UVA + UVB (PAR + UVA + 280– 320 nm). Sublittoral thalli collected around Helgoland, North Sea, Germany, from 6 m below the mean low water of spring tides contained less than 0.1 mg·g⁻¹ dry weight (DW) total MAAs, whereas eulittoral samples contained over 1 mg·g⁻¹ DW. Transplantation to shallow water led to the immediate synthesis of three MAAs in the following temporal order: shinorine (λ_max 334 nm), asterina (λ_max 330 nm), and palythine (λ_max 320 nm), with the shinorine content peaking and then declining after 2 days (exposure to 100 mol photons·m⁻²). Maximum total MAA content (2 mg·g⁻¹ DW) also occurred after 2 days of induction, exceeding the content normally found in eulittoral samples. Furthermore, the relative proportion of the different MAAs at this time was different than that in eulittoral samples. After 2 days the total content declined to the eulittoral value, with palythine as the principal MAA. Similar data were obtained for all treatments, indicating that MAA synthesis in C. crispus was induced by PAR and not especially stimulated by UV radiation. The ability of photosystem II (PSII) to resist damage by UVB was tested periodically during the acclimation period by exposing samples to a defined UVB dose in the lab. Changes in chlorophyll fluorescence (F_v/F_m and effective quantum yield, φ_II) indicated that PSII function was inhibited during the initial stage of acclimation but gradually improved with time. No difference among screening treatments was detected except in spring for the samples acclimating to PAR + UVA + UVB. In this treatment F_v/F_m and φ_II were significantly lower than in the other treatments. During the first week of each experiment, growth rates were also significantly reduced by UVB. The reductions occurred despite maximum MAA content, indicating an incomplete protection of photosynthetic and growth-related processes.     

EFFECT OF HYPOTHERMIA UPON ORGANIC PHOSPHATES, GLYCOLYTIC METABOLITES, CITRIC ACID CYCLE INTERMEDIATES AND ASSOCIATED AMINO ACIDS IN RAT CEREBRAL CORTEX

—The influence of hypothermia upon the metabolism of the brain was studied by reducing body temperature in N₂O-anaesthetized rats to 32, 27 or 22°C, with subsequent measurements of organic phosphates, glycolytic metabolites, citric acid cycle intermediates and associated amino acids. Hypothermia was maintained for either 1 or 2 h and the effect of anaesthesia was evaluated by maintaining unanaesthetized animals at 22°C. Hypothermia had no influence on the cerebral cortical concentrations of ATP, ADP or AMP and there was only a small increase in phosphocreatine. Since the tissue concentrations of glucose and glycogen were reduced, it is concluded that the well known resistance of the hypothermie brain to ischaemia is unrelated to increased energy stores. Hypothermia was accompanied by decreases in the tissue concentrations of fructose-1,6-diphosphate, dihydroxyacetone phosphate, 3-phosphoglycerate, pyruvate, lactate, α-ketoglutarate, succinate and malate, but not of glucose-6-phosphate or citrate. These results indicate that metabolic flux is retarded mainly at the phosphofructokinase and isocitrate dehydrogenase steps. The largest relative reduction was seen in α-ketoglutarate, which was possibly secondary to accumulation of ammonia. There was no change in GABA, but a decrease in glutamate and increases in aspartate and alanine. These, changes are compatible with shifts in the aspartate and alanine aminotransferase reactions, possibly induced by the fall in α-ketoglutarate.     

ENHANCEMENT OF INTRACELLULAR PROLINE LEVEL IN CELLS OF ANACYSTIS NIDULANS (CYANOBACTERIA) EXPOSED TO DELETERIOUS CONCENTRATIONS OF COPPER1

The intracellular proline level in Anacystis nidulans cells was enhanced when the cells were exposed to sublethal concentrations of Cu²⁺; the degree of enhancement was positively related to the concentration of Cu²⁺. Analysis by high-performance liquid chromatography confirmed that the enhancement of proline levels was the most pronounced change in the composition of the free amino acid pool during copper treatment. A direct supply of exogenous proline to the cultures lowered the inhibitory influence of Cu²⁺on the growth of cells. Further experiments showed that the supply of exogenous proline lowered the leakage of potassium ions from cells exposed to deleterious concentrations of Cu²⁺. The inhibition of potassium leakage was particularly pronounced when proline was supplied prior to Cu²⁺treatment. The present study suggests that enhanced proline protects cell membranes from being affected by deleterious concentrations of Cu²⁺.