Abundance and composition of the summer phytoplankton community along a transect from the Barguzin River to the central basin of Lake Baikal

The abundance and composition of phytoplankton were investigated at six stations along a transect from the Barguzin River inflow to the central basin of Lake Baikal in August 2002 to clarify the effect of the river inflow on the phytoplankton community in the lake. The water temperature in the epilimnion was high near the shore at Station 1 (17.3°C), probably due to the higher temperature of the river water, and gradually decreased offshore at Station 6 (14.5°C). Thermal stratification developed at Stations 2–6, and a thermocline was observed at a 17–22-m depth at Stations 2–4 and an 8–12-m depth at Stations 5 and 6. The concentrations of nitrogen and phosphorus nutrients in the epilimnion at all stations were <1.0 μmol N l⁻¹ and <0.16 μmol P l⁻¹, respectively. Relatively high concentrations of nutrients (0.56–7.38 μmol N l⁻¹ and 0.03–0.28 μmol P l⁻¹) were detected in the deeper parts of the euphotic zone. Silicate was not exhausted at all stations (>20 μmol Si l⁻¹). The chlorophyll a (chl. a) concentration was high (>10 μg l⁻¹) near the shore at Station 1 and low (<3 μg l⁻¹) at five other stations. The <2 μm fraction of chl. a in Stations 2–6 ranged between 0.80 and 1.85 μg l⁻¹, and its contribution to total chl. a was high (>60%). In this fraction, picocyanobacteria were abundant at all stations and ranged between 5 × 10⁴ and 5 × 10⁵ cells ml⁻¹. In contrast, chl. a in the >2 μm fraction varied significantly (0.14–11.17 μg l⁻¹), and the highest value was observed at Station 1. In this fraction, the dominant phytoplankton was Aulacoseira and centric diatoms at Station 1 and Cryptomonas, Ankistrodesmus, Asterionella, and Nitzschia at Stations 2–6. The present study demonstrated the dominance of picophytoplankton in the pelagic zone, while higher abundance of phytoplankton dominated by diatoms was observed in the shallower littoral zone. These larger phytoplankters in the littoral zone probably depend on nutrients from the Barguzin River.     

Thidiazuron-induced high-frequency direct shoot organogenesis of <Emphasis Type="Italic">Cannabis sativa</Emphasis> L.

Induction of high-frequency shoot regeneration using nodal segments containing axillary buds from a 1-yr-old mother plants of Cannabis sativa was achieved on Murashige and Skoog (MS) medium containing 0.05–5.0 μM thidiazuron. The quality and quantity of regenerants were better with thidiazuron (0.5 μM thidiazuron) than with benzyladenine or kinetin. Adding 7.0 μM of gibberellic acid into a medium containing 0.5 μM thidiazuron slightly increased shoot growth. Elongated shoots when transferred to half-strength MS medium supplemented with 500 mg l⁻¹ activated charcoal and 2.5 μM indole-3-butyric acid resulted in 95% rooting. The rooted plants were successfully acclimatized in soil. Following acclimatization, growth performance of 4-mo-old in vitro propagated plants was compared with ex vitro vegetatively grown plants of the same age. The photosynthesis and transpiration characteristics were studied under different light levels (0, 500, 1,000, 1,500, or 2,000 μmol m⁻² s⁻¹). An increase in photosynthesis was observed with increase in the light intensity up to 1,500 μmol m⁻² s⁻¹ and then decreased subsequently at higher light levels in both types of plants. However, the increase was more pronounced at lower light intensities below 500 μmol m⁻² s⁻¹. Stomatal conductance and transpiration increased with light intensity up to highest level (2000 μmol m⁻² s⁻¹) tested. Intercellular CO₂ concentration (C _i) and the ratio of intercellular CO₂ concentration to ambient CO₂ (C _i/C _a) decreased with the increase in light intensity in both in vitro as well as ex vitro raised plants. The results show that in vitro propagated and hardened plants were functionally comparable to ex vitro plants of same age in terms of gas and water vapor exchange characteristics, within the limits of this study.     

Concentration dependent growth/non-growth linked kinetics of endosulfan biodegradation by <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>

The rates of biodegradation of endosulfan by P. aeruginosa were determined with different initial endosulfan concentrations (10, 50, 100, 150, 200 and 250 mg l⁻¹) and different growth linked kinetic models were fitted at these concentrations. At 10 mg endosulfan l⁻¹, Monod no growth model was well fitted. Monod with growth model described the biodegradation pattern at an initial concentration of 50, 100 and 150 mg endosulfan l⁻¹. Significant increases of P. aeruginosa MN2B14 density in broth culture during incubation further support this result. Conversely, zero order kinetic model was well fitted into the biodegradation data if initial endosulfan concentration was ≥200 mg endosulfan l⁻¹. The kinetics of endosulfan biodegradation by P. aeruginosa MN2B14 in liquid broth was highly dependent upon its initial concentration. The results of this study could be employed for predicting the persistence of endosulfan in water environment containing P. aeruginosa as an endosulfan degrading bacterium.     

Physiological differences in the growth of <Emphasis Type="Italic">Sargassum horneri</Emphasis> between the germling and adult stages

The effects of temperature, irradiance, and daylength on Sargassum horneri growth were examined at the germling and adult stages to discern their physiological differences. Temperature–irradiance (10, 15, 20, 25, 30°C × 20, 40, 80 μmol photons m⁻²s⁻¹) and daylength (8, 12, 16, 24 h) experiments were carried out. The germlings and blades of S. horneri grew over a wide range of temperatures (10–25°C), irradiances (20–80 μmol photons m⁻²s⁻¹), and daylengths (8–24 h). At the optimal growth conditions, the relative growth rates (RGR) of the germlings were 21% day⁻¹ (25°C, 20 μmol photons m⁻²s⁻¹) and 13% day⁻¹ (8 h daylength). In contrast, the RGRs of the blade weights were 4% day⁻¹ (15°C, 20 μmol photons m⁻²s⁻¹) and 5% day⁻¹ (12 h daylength). Negative growth rates were found at 20 μmol photons m⁻²s⁻¹ of 20°C and 25°C treatments after 12 days. This phenomenon coincides with the necrosis of S. horneri blades in field populations. In conclusion, we found physiological differences between S. horneri germlings and adults with respect to daylength and temperature optima. The growth of S. horneri germlings could be enhanced at 25°C, 20 μmol photons m⁻²s⁻¹, and 8 h daylength for construction of Sargassum beds and restoration of barren areas.     

Effects of nutrients on growth of the red-tide dinoflagellate <Emphasis Type="Italic">Gyrodinium instriatum</Emphasis> Freudenthal et Lee and a possible link to blooms of this species

We investigated the impact of different nitrogen (N) and phosphorus (P) compounds and concentrations on the growth of Gyrodinium instriatum Freudenthal et Lee in laboratory experiments, and possible links to blooms of this species at Hakozaki Fishing Port, Fukuoka, Japan. G. instriatum utilized only inorganic N compounds as N sources for growth. In contrast, G. instriatum utilized many inorganic and organic phosphorus compounds. We used the Monod equation to describe the growth rate of G. instriatum in N- or P-limited batch cultures as a function of ambient nutrient concentrations. Kinetic growth parameters for maximum specific growth rate (μ_max) and half-saturation nutrient concentration (K _S) were 0.57 divisions d⁻¹ and 14.2 μmol l⁻¹, respectively, under N-limitation and 0.65 divisions d⁻¹ and 1.75 μmol l⁻¹, respectively, under P-limitation. Compared with these K _S values, all in situ average dissolved inorganic nitrogen (DIN) concentrations in Hakozaki Fishing Port were higher than K _S for N, but all in situ average dissolved inorganic phosphorus (DIP) concentrations were lower than K _S for P, whether a red tide occurred or not bloom. Moreover, average DIP concentration in April (a month critical to red-tide genesis) of 2004 (a non-red-tide year) was less than half those in 2002 and 2003 (red-tide years). Thus, differences in DIP concentrations may be an important factor controlling blooms of G. instriatum in Hakozaki Fishing Port.     

Growth retardants stimulate guggulsterone production in the presence of fungal elicitor in fed-batch cultures of <Emphasis Type="Italic">Commiphora wightii</Emphasis>

Guggulsterone, a hypolipidemic natural agent, is produced in resin canals of the plant Commiphora wightii. In this study, the stimulatory effects of growth retardants [ALAR (N,N-dimethylaminosuccinamic acid) and CCC (chlormequat chloride)] and fungal elicitor on guggulsterone accumulation in cell cultures of C. wightii are reported. CCC at 1 mg l⁻¹ enhanced guggulsterone content (~123 μg l⁻¹) when added on the fifth day after inoculation, while ALAR at 2.5 mg l⁻¹ increased guggulsterone content (~116 μg l⁻¹) when added on the tenth day. In a two-stage fed-batch process, combined treatment with fungal elicitor and growth retardant caused a significant increase (~353 μg l⁻¹) in guggulsterone content in cell cultures after 17 days of growth. This represents an approximately fivefold increase over the guggulsterone contents in initial cultures of this plant.     

Heterotrophic production of eicosapentaenoid acid by the diatom Nitzschia laevis: effects of silicate and glucose

The effects of silicate and glucose on growth and eicosapentaenoic acid (EPA) production by the diatom Nitzschia laevis were studied. By alternately altering the concentrations of silicate (2.7–64 mg l⁻¹) and glucose (1–40 g l⁻¹) in the medium, the highest cell dry weight (ca. 5.5 g l⁻¹) was obtained at 20 g l⁻¹ glucose and 32 mg l⁻¹ silicate, while the highest specific growth rate (ca. 0.65 day⁻¹) was obtained at a relatively low glucose concentration (5 g l⁻¹) and high silicate concentrations (32–64 mg l⁻¹). At glucose levels of 5 and 20 g l⁻¹, EPA content was higher with lower silicate concentrations (2.7 and 16 mg l⁻¹ silicate, respectively), while at a silicate level of 16 mg l⁻¹, higher glucose concentrations (20–40 g l⁻¹) facilitated EPA formation. The highest EPA yield (131 mg l⁻¹) was obtained at 20 g l⁻¹ glucose and 32 mg l⁻¹ silicate, while the highest EPA productivity (15.1 mg l⁻¹ day⁻¹) was obtained at 20 g l⁻¹ glucose and 64 mg l⁻¹ silicate. Journal of Industrial Microbiology & Biotechnology (2000) 25, 218–224. Received 08 May 2000/ Accepted in revised form 21 July 2000     

CO<Subscript>2</Subscript>-enriched microenvironment affects sucrose and macronutrients absorption and promotes autotrophy in the <Emphasis Type="Italic">in vitro</Emphasis> culture of kiwi (<Emphasis Type="Italic">Actinidia deliciosa</Emphasis> Chev. Liang and Ferguson)

In traditional in vitro culture, the low CO₂ concentration inside the vessels restricts photosynthesis and necessitates the addition of sucrose to the culture medium as the main energy source, thus bringing about changes in the absorption of mineral elements from the culture medium. In this study, we investigated macronutrient absorption and sugar consumption in Actinidia deliciosa Chevalier Liang and Ferguson cv. Hayward (kiwi), cultured on medium supplemented with varying amounts of sucrose (0, 10, and 20 g l⁻¹) under both heterotrophy and autotrophy, flushed with different concentrations of CO₂ (non-ventilation, 300, 600, and 2,000 μl l⁻¹). In ventilated systems with 20 g l⁻¹ of sucrose, sucrose absorption was less than under non-ventilation. The lowest rate of sucrose absorption was recorded when the explants were cultured on medium supplemented with 20 g l⁻¹ of sucrose and flushed with 600 μl l⁻¹ CO₂. Absorption of NO₃ ⁻, PO₄ ³⁻, and Mg²⁺ were high (maximum) at the end of the culture period (40 d) in explants flushed with 600 μl l⁻¹ CO₂ that have been cultured 20 d in the presence of sucrose and then transferred to a sucrose-free medium. These autotrophic conditions promoted maximum plant growth in terms of both fresh and dry mass as well as the length and number of shoots and leaves. The study shows that to maintain an optimum regime of mineral nutrition for prolonged culture of kiwi in vitro, an increased amount of these three ions should be supplemented in Murashige and Skoog’s medium.     

Seasonal variations of microphytoplankton assemblages and environmental variables in the coastal zone of Bozcaada Island in the Aegean Sea (NE Mediterranean Sea)

Seasonal changes in the microphytoplankton assemblages were examined in the coastal zone of Bozcaada Island with regard to some major physical and chemical variables. Samples were collected from May 2000 to December 2001 at four stations. A total of 108 dinoflagellates, 102 diatoms, 1 chrysophycean, 3 dictyochophycean, and 1 prasinophycean species were identified and quantified during the study period. Diatoms and dinoflagellates were the most important in terms of species number and abundance. The maximum values of total microphytoplankton were observed at 0.5 m depth (46.2 × 10³ cells l⁻¹ at st. 3) in May as this was the month when the diatom Pseudo-nitzschia pungens bloomed. Chlorophyll (chl) a concentration ranged between 0.08 (August) and 0.78 μg l⁻¹ (February). May was another important month in which chlorophyll a increased (0.41–0.47 μg l⁻¹). Species diversity values (H′_log2) ranged from 1.66 bits (June, 20 m) to 4.11 bits (November, 0.5 m). The increase was attributed to a more balanced distribution of abundance among species. The amounts of nitrate + nitrite (0.6−3.7 μg-at N l⁻¹), phosphate (0.2−0.6 μg-at P l⁻¹) and silicate (0.7−2.5 μg-at Si l⁻¹) were recorded on each sampling occasion. Nutrient concentrations and chl a values of the research area were found to be poorer than those of the many other coastal areas in the northeastern Mediterranean. The mean atomic ratio of nitrogen to phosphorus varied from 1.3 (June) to 12.9 (February). This ratio was lower than the Redfield ratio of 16 for ocean phytoplankton, and phytoplankton was potentially limited by nitrogen for most of the months. The result of this study confirms and emphasizes the oligotrophic nature of the eastern Mediterranean.     

Enhanced extracellular production of trans-resveratrol in Vitis vinifera suspension cultured cells by using cyclodextrins and methyljasmonate

In this work, the effect of different inducing factors on trans-resveratrol extracellular production in Monastrell grapevine suspension cultured cells is evaluated. A detailed analysis provides the optimal concentrations of cyclodextrins, methyljasmonate and UV irradiation dosage, optimal cell density, elicitation time and sucrose content in the culture media. The results indicate that trans-resveratrol production decreases as the initial cell density increases for a constant elicitor concentration in Monastrell suspension cultured cells treated with cyclodextrins individually or in combination with methyljasmonate; the decrease observed in cell cultures elicited with cyclodextrins alone is far more drastic than those observed in the combined treatment. trans-Resveratrol extracellular production observed by the joint use of cyclodextrins and methyljasmonate (1,447.8 ± 60.4 μmol trans-resveratrol g⁻¹ dry weight) is lower when these chemical compounds are combined with UV light short exposure (669.9 ± 45.2 μmol trans-resveratrol g⁻¹ dry weight). Likewise, trans-resveratrol production is dependent on levels of sucrose in the elicitation medium with the maximal levels observed with 20 g l⁻¹ sucrose and the joint action of cyclodextrins and 100 μM methyljasmonate. The sucrose concentration did not seem to limit the process although it affects significantly the specific productivity since the lowest sucrose concentration is 10 g l⁻¹, the highest productivity is reached (100.7 ± 5.8 μmol trans-resveratrol g⁻¹ dry weight g⁻¹ sucrose) using cyclodextrins and 25 μM methyljasmonate.     

Long-term production of soluble human Fas ligand through immobilization of <Emphasis Type="Italic">Dictyostelium discoideum</Emphasis> in a fibrous bed bioreactor

The production of recombinant glycoproteins in Dictyostelium discoideum by conventional cell culture methods was limited by low cell density as well as low growth rate. In this work, cotton towel with a good adsorption capability for D. discoideum cells was used as the immobilization matrix in an external fibrous bed bioreactor (FBB) system. With batch cultures in the FBB, the concentration of immobilized cells in the cotton fiber carrier increased to 1.37 × 10⁸ cells per milliliter after 110-h cultivation, which was about tenfold higher than the maximal cell density in the conventional free-cell culture. Correspondingly, a high concentration of soluble human Fas ligand (hFasL; 173.7 μg l⁻¹) was achieved with a high productivity (23 μg l⁻¹ h⁻¹). The FBB system also maintained a high density of viable cells for hFasL production during repeated-batch cultures, achieving a productivity of 9∼10 μg l⁻¹ h⁻¹ in all three batches studied during 15 days. The repeated-batch culture using immobilized cells of D. discoideum in the FBB system thus provides a good method for long-term and high-level production of hFasL.     

Indirect regeneration of the Cancer bush (<Emphasis Type="Italic">Sutherlandia frutescens</Emphasis> L.) and detection of <Emphasis Type="SmallCaps">l</Emphasis>-canavanine in <Emphasis Type="Italic">in vitro</Emphasis> plantlets using NMR

The present study reports a simple protocol for indirect shoot organogenesis and plant regeneration of Sutherlandia using rachis and stem segments. Different concentrations (0.0–68.08 μmol l⁻¹) of thidiazuron (TDZ) were used for callus induction and shoot organogenesis. The highest percentage of callus formation (97.5%) and the highest percentage of explants forming shoots (88.8%) were obtained from rachis explants cultured onto Murashige and Skoog (MS) medium (Murashige and Skoog, Physiol. Plant. 15:473–495, 1962) supplemented with 45.41 μmol l⁻¹ TDZ. Scanning electron microscopy demonstrated the early development of adventitious shoots derived from callus cultures. Shoot clusters were further developed and grown in MS hormone-free medium. The presence of l-canavanine was determined by thin-layer chromatography and confirmed after column fractionation using silica gel and nuclear magnetic resonance spectroscopy. Individual shoots were rooted on different concentrations and combinations of MS salt strength and IBA. Half-strength MS salt medium supplemented with 24.6 μmol l⁻¹ IBA was optimal for root induction in which 78% of shoots were rooted. The in vitro plants were successfully acclimatized in a growth chamber with a 90% survival rate.     

Critical limits for acid neutralizing capacity of brown trout (<Emphasis Type="Italic">Salmo trutta</Emphasis>) in Norwegian lakes differing in organic carbon concentrations

A regional data set on water chemistry from 1995 was used to set critical values for the survival of brown trout in Norwegian lakes (n = 790) in relation to pH, inorganic Al and acid neutralizing capacity (ANC). ANC was estimated both traditionally (ANC_trad) and modified by treating one-third of the dissolved organic matter as part of the strong acid anions (ANC_mod). The threshold value to avoid fish damage (ANC_limit) was compared with that found in a similar study from 1986. Brown trout populations were categorized as unaffected, damaged or extinct on the basis of questionnaires. In 1995, threshold values to avoid fish damage on the basis of ANC_trad and ANC_mod were 67 and 48 μeq l⁻¹, respectively, compared with 20 and 8 μeq l⁻¹, respectively, in 1986. The higher ANC_limit found for the data from 1995 is probably caused by a lower pH and a higher inorganic Al concentration at a given ANC value in 1995 than in the 1980s. ANC_limit was highly related to organic carbon concentrations in the study lakes, being estimated at 33, 73 and >100 μeq l⁻¹ for three different TOC categories (<2 mg C l⁻¹, 2–5 mg C l⁻¹ and >5 mg C l⁻¹). These differences in ANC_limit are due to lower pH and higher concentrations of inorganic Al in humic lakes than in clear water lakes at the same level of ANC. It is suggested that the change in ANC_limit for fish in acidified lakes is linked to increased concentrations of TOC in recent years.     

Stimulation of menthol production in <Emphasis Type="Italic">Mentha piperita</Emphasis> cell culture

Plant cell culture provides an alternative means for producing secondary metabolites. In this study, experiments were carried out to study the impact of several parameters, independently and in combination, on the stimulation of menthol production in the cell suspension culture of Mentha piperita. Callus was obtained from leaf segments of in vitro grown plantlets on Murashige and Skoog (MS) medium supplemented with 0.2 mg l⁻¹of 2,4-dichlorophenoxy acetic acid to initiate cell suspension culture. This culture was maintained in half-strength MS medium supplemented with 0.2 mg l⁻¹of 2,4-dichlorophenoxy acetic acid at 15 d interval and used for further studies. Precursor feeding alone, i.e., menthone, at 35 μM concentration showed slightly improved productivity. γ-Cyclodextrin alone at 60 μM concentration and in combination with menthone feeding at 35 μM increased menthol yield up to 92 and 110 mg l⁻¹ in comparison to 77 mg l⁻¹ of control culture. Synergistic potentiation effect of menthone feeding at 35 μM and γ-cyclodextrin at 60 μM treatment followed by in situ adsorption with RP-8 also showed potential stimulation of menthol production in M. piperita cell culture. Fungal elicitor treatment showed enhanced production level up to 140.8 mg l⁻¹ in comparison to that of control. Further studies were carried out with the establishment of Agrobacterium tumefaciens (Ach5) gall-mediated calli, and consequently, cell suspension culture and results showed the significant enhancement of menthol yield up to 278 mg l⁻¹. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Plant regeneration via somatic embryogenesis of <Emphasis Type="Italic">Camptotheca acuminata</Emphasis> in temporary immersion system (TIS)

The present study describes a protocol for plant regeneration via somatic embryogenesis in temporary immersion system (TIS) for Camptotheca acuminata. Somatic embryos were induced by culturing hypocotyl segments from 14-day-old in vitro grown C. acuminata seedlings in TIS. Hypocotyl segments were placed in culture vessels modified with a mechanical device to support the fixation of explants. Cultures were maintained under a 16 h photoperiod with a light intensity of 60 μmol m⁻² s⁻¹ PPF at 25 ± 1°C. After 16 weeks of incubation embryogenic calli were formed above the edge of the mechanical device in the basal Murashige and Skoog (MS) medium containing 35 g l⁻¹ sucrose and without hormonal supplementation. For plantlet regeneration, somatic embryos at cotyledonary stage were cultured in three different concentrations of 6-benzylamino-purine (0.5, 1.0 and 1.5 mg l⁻¹ BAP) and in plant growth regulator (PGR) free medium. In general, 0.5 mg l⁻¹ BAP was found to be the most effective concentration for growth and development of Camptotheca embryos in TIS. Conversion of somatic embryos into plantlets was also successfully achieved on sterile substrates moistened with 0.5 mg l⁻¹ BAP. Plantlets derived from cotyledonary embryos were rooted in vitro with 0.5 mg l⁻¹ indole-3-butyric acid (IBA) before transfer to ex vitro conditions.     

Effect of irradiation intensity on cell growth and kalata B1 accumulation in <Emphasis Type="Italic">Oldenlandia affinis</Emphasis> cultures

Light irradiation had remarkable effects on callus growth of Oldenlandia affinis with an optimum intensity of 35 μmol m⁻² s⁻¹. Biosynthesis of kalata B1, the main cyclic peptide in O. affinis, was induced and triggered with rising irradiation intensities. The highest concentration of kalata B1, 0.49 mg g⁻¹ DW characterised by the maximum productivity of 3.88 μg per litre and day was analysed at 120 μmol m⁻² s⁻¹, although callus growth was repressed. The light saturation point was established to be 35 μmol m⁻² s⁻¹, where kalata B1 productivity was in a similar order (3.41 μg per day) due to the higher growth index. O. affinis suspension cultures were shown to accumulate comparable specific kalata B1 concentrations in a delayed growth associated production pattern. These were dependent on irradiation intensity (0.16 mg g⁻¹ at 2 μmol m⁻² s⁻¹; 0.28 mg g⁻¹ at 35 μmol m⁻² s⁻¹). The batch cultivation process resulted in a maximum productivity of 27.30 μg per litre and day with culture doubling times of 1.16 d⁻¹. Submers operation represented a 8-fold product enhancement compared to callus cultivation.     

Physical and chemical limnology of a wind-swept tropical highland reservoir

Valle de Bravo (VB) is a tropical reservoir located (19°21′30″ N, 100°11′00″ W) in the highlands of Mexico. The reservoir is daily swept by strong (7.4 m s⁻¹ mean speed) diurnal (12:00–19:00 h) winds that blow along its two main arms. As expected from its fetch (6.9 km) and its depth (21.1 m mean), the reservoir behaves as a warm monomictic water body. During 2001, VB was stratified from February to October, and well mixed from November to January. Its mean temperature was 19.9°C; the maximum found was 23.8°C in the epilimnion, while a minimum of 17.8°C was registered during mixing. VB exhibited a thermal regime similar to other water bodies of the Mexican tropical highlands, except for a steady increase of its hypolimnetic temperature during stratification, which is attributed to entrainment of epilimnetic water into the hypolimnion. During stratification, the hypolimnion was anoxic, while the whole water column remained under-saturated (60%) during mixing. The flushing time is 2.2 years. Mineralization and total alkalinity are low, which allows strong changes in pH. Ammonia remained low (2.4 μmol l⁻¹ mean) in the epilimnion, but reached up to 60 μmol l⁻¹ in the hypolimnion. Soluble reactive phosphorous had a mean of 0.28 μmol l⁻¹ in the epilimnion and a mean of 1.25 μmol l⁻¹ in the hypolimnion. Nitrate exhibited maxima (up to 21 μmol l⁻¹) during mixing, and also in the metalimnion (2 μmol l⁻¹) during stratification. Low dissolved inorganic nitrogen indicated nitrogen limitation during stratification. Eutrophication is an emerging problem in VB, where cyanobacteria dominate during stratification. At VB chlorophyll a is low during mixing (mean of 9 μg l⁻¹), and high during stratification (mean 21 μg l⁻¹), when blooms (up to 88 μg l⁻¹) are frequent. This pattern is similar to that found in other eutrophic tropical water bodies. We propose that in VB the wind regime causes vertical displacements of the thermocline (0.58–1.10 m hr⁻¹) and boundary mixing, enhancing the productivity during the stratification period in this tropical reservoir.     

Eco-physiological responses of nitrogen-fixing cyanobacteria to light

The eco-physiological responses of three nitrogen-fixing cyanobacteria (N-fixing cyanobacteria), Aphanizomenon gracile, Anabaena minderi, and Ana. torques-reginae, to light were assessed under nutrient saturation. The N-fixing cyanobacteria were isolated into monocultures from a natural bloom in a shallow colored lake and their growth irradiance parameters and pigment composition were assessed. The different ecological traits related to light use (μ_max, α, I _k) suggest that these N-fixing cyanobacteria are well adapted to low light conditions at sufficient nutrients, yet interspecific differences were observed. Aphanizomenon gracile and Anabaena minderi had high relative growth rates at low irradiances (ca. 70% of those in high light), low half saturation constant for light-limited growth (I _k < 9.09 μmol photon m⁻² s⁻¹) and high efficiency (α < 0.11 day⁻¹ μmol photon⁻¹ m² s). Conversely, Ana. torques-reginae showed poorer light competitiveness: low relative growth rates at low irradiances (ca. 40% of those in high light), low α (0.009 day⁻¹ μmol photon⁻¹ m² s) and higher I _k (35.5 μmol photon m⁻² s⁻¹). Final densities in Aphanizomenon gracile and Anabaena minderi reached bloom densities at irradiances above 30 μmol photon m⁻² s⁻¹ with different hierarchy depending on irradiance, whereas Ana. torques-reginae never achieved bloom densities. All species had very low densities at irradiances ≤17 μmol photon m⁻² s⁻¹, thus no N-fixing blooms would be expected at these irradiances. Also, under prolonged darkness and at lowest irradiance (0 and 3 μmol photon m⁻² s⁻¹) akinetes were degraded, suggesting that in ecosystems with permanently dark sediments, the prevalence of N-fixing cyanobacteria should not be favored. All species displayed peaks of phycocyanin, but no phycoeritrin, probably due to the prevailing red light in the ecosystem from which they were isolated.     

Effects of salinity on growth and compatible solutes of callus induced from <Emphasis Type="Italic">Populus euphratica</Emphasis>

Summary The present study aimed to evaluate the response to salinity of Populus euphratica, which is more salt-resistant than other poplar cultivars, at the cellular level. To this purpose, callus was induced from shoot segments of P. euphratica on Murashige and Skoog (MS) medium supplemented with 0.5 mg l⁻¹ (2.2 μM) 6-benzyladenine (BA) and 0.5 mg l⁻¹ (2.7 μM 1-naphthaleneacetic acid (NAA). Callus was transferred to MS medium supplemented with 0.25 mg l⁻¹ (1.1 μM) BA and 0.5 mg l⁻¹ NAA. The relative growth rate of callus reached a maximum in the presence of 50 mmol l⁻¹ NaCl and growth was inhibited with increasing NaCl concentrations. Examination of the changes of osmotic substances under salt stress showed that accumulation of proline, glycine betaine, and total soluble sugars increased with increasing salt concentrations. The results indicate that the response of the callus of P. euphratica to salt stress is similar to that of the whole plant.     

Photolimitation and photoinhibition as factors determining optimal dilution rate to produce eicosapentaenoic acid from cultures of the microalga Isochrysis galbana

Eicosapentaenoic acid (EPA) productivity from continuous cultures of the marine microalga Isochrysis galbana was studied, taking into account the irradiance on the reactor surface, that is, the photolimitation/photoinhibition regime to which the cells are exposed. Experiments were conducted under a wide variety of operating conditions. The dilution rate ranged from 0.005 h⁻¹ to 0.040 h⁻¹ at five external irradiances (820, 1620, 2050, 2450 and 3270 μmol photons m⁻² s⁻¹) covering photolimited to photoinhibited growth. Under these conditions, the specific growth rate (μ) was found to be the main factor influencing EPA content (ranging from 2.35% to 5.23% dryweight) and productivity (up to 0.88 mg l⁻¹ h⁻¹). The fatty acid content was not significantly affected by the external irradiance, but was influenced by the state of growth of the microalga, depending on whether the light regime was photolimiting or photoinhibiting. It might be suggested that light should no longer be considered an isolated factor affecting EPA synthesis, but an indirect influence through the photolimitation/photoinhibition regime and growth rate. At a given dilution rate, EPA content and biomass concentration are lower under photoinhibiting external irradiances than those corresponding to photolimiting conditions, and consequently EPA productivity decays. Since the effect of photoinhibition is less marked at high biomass concentration, a strategy to optimize EPA productivity from microalgal cultures could consist of reducing the dilution rate when the external irradiance increases above the phoinhibition threshold. Received: 16 January 1998 / Revised revision: 27 March 1998 / Accepted: 27 March 1998