Kinetics study of pyridine biodegradation by a novel bacterial strain,Rhizobium sp. NJUST18

Biodegradation of pyridine by a novel bacterial strain, Rhizobium sp. NJUST18, was studied in batch experiments over a wide concentration range (from 100 to 1,000 mg l^?1). Pyridine inhibited both growth of Rhizobium sp. NJUST18 and biodegradation of pyridine. The Haldane model could be fitted to the growth kinetics data well with the kinetic constants μ* = 0.1473 h^?1, K _s = 793.97 mg l^?1, K _i = 268.60 mg l^?1 and S _m = 461.80 mg l^?1. The true μ _max, calculated from μ*, was found to be 0.0332 h^?1. Yield coefficient Y _X/S depended on S _i and reached a maximum of 0.51 g g^?1 at S _i of 600 mg l^?1. V _max was calculated by fitting the pyridine consumption data with the Gompertz model. V _max increased with initial pyridine concentration up to 14.809 mg l^?1 h^?1. The q _S values, calculated from $V_{ \hbox{max} }$ , were fitted with the Haldane equation, yielding q _Smax = 0.1212 g g^?1 h^?1 and q* = 0.3874 g g^?1 h^?1 at S _m′ = 507.83 mg l^?1, K _s′ = 558.03 mg l^?1, and K _i′ = 462.15 mg l^?1. Inhibition constants for growth and degradation rate value were in the same range. Compared with other pyridine degraders, μ _max and S _m obtained for Rhizobium sp. NJUST18 were relatively high. High K _i and K _i′ values and extremely high K _s and K _s′ values indicated that NJUST18 was able to grow on pyridine within a wide concentration range, especially at relatively high concentrations.     

Rapid treatment of vessels fouled with an invasive polychaete,Sabella spallanzanii,using a floating dock and chlorine as a biocide

Chlorine solution was added to the water encapsulated within a proprietary ‘floating dock’ to treat a vessel infested with the invasive polychaete Sabella spallanzanii. The chlorine was added as sodium dichloroisocyanurate (‘dichlor’) at an initial concentration of 200 mg l^?1 of free available chlorine (FAC). This concentration killed 99% of S. spallanzanii in their tubes during a 4-h exposure in laboratory tests (EC₉₉ 160 mg FAC l^?1). The concentration of FAC in the floating dock declined to ~50 mg l^?1 after 4 h and < 10 mg l^–1 after 16 h. Residual FAC was neutralised with thiosulphate at completion of exposure. A sample of 30 S. spallanzanii individuals collected from the hull after treatment all showed morphological damage and 28 showed no response to touch. Re-examination of the hull after 6 d found no live worms or other fouling organisms. This method provides a cost-effective, rapid means of treating hull fouling.     

LEAD INHIBITION OF CHLOROPHYCEAN MICROALGAE1

The effects of lead on the growth of 9 microalgae representing 4 chlorophycean genera were investigated in axenic laboratory cultures. Variations in sensitivity to lead existed among the algae tested with availability of phosphate often limiting growth. Under conditions in which phosphate was growth limiting at pH 6.2, Chlorella pyrenoidosa Chick, C. ellipsoidea Gerneck, C. vulgaris Beij. Scenedesmus sp. (IUCC 1591).Sc. Obtusiusculus Chodat and Selenastrum capricornutum Printz first showed toxicity at 0.5 mg·l^?1 pb. Two planktonic Scenedesmus spp. And an Ankistrodesmus sp. Were more tolerant, developing toxicity at 1 mg· l^?1 Pb. The relationship of phosphate supply to Pb toxicity was investigated with S. capricornutum and Sc. Obtusiusculus. Al pH 8.0, Selenastrum developed toxicity at 3 mg·l^?1 pb either with or without phosphate. When phosphate supply was growth-limiting, 6 times as much pb (i.e., 3 vs. 0.5 mg l^?1 was required to demonstrate toxicity with Selenastrum at pH 8.0 as opposed to pH 6.2 Sc. Obtusiusculus was inhibited at 0.5 mg· l^?1 pb in phosphate-limiting medium and developed toxicity at 1 mg·l^?1 in media containing phosphate. Control of colonial morphology with Sc. Obtusiusculus was inhibited. Phosphate availability as well as pH would seem to be important factors determining lead toxicity with some algae.     

Some physical and chemical characteristics of Lake Atnsjøen

The physical and chemical characteristics of the dimictic, ultraoligotrophic, and subalpine Lake Atnsjøen are described based on data from the period 1985–2001. The temperature stratification of the lake is weakly developed, with the thermocline at about 10 m depth. The surface temperature during summer is usually lower than 14–15?°C. The Secchi disk transparency is normally higher than 8–10 m in summer and autumn, but can be as low as 4.3 m during spring. The pH is about 6.0, and the specific conductivity about 1.0 mS m^{? 1}. The dominating cation and anion are Ca (≈0.8 mg l^?1 or 42 μeq l^?1), and SO₄ (≈1.5 mg l^?1 or 33 μeq l^?1), respectively. The concentration of bicarbonate (HCO₃, μeq l^?1) is sometimes lower, sometimes higher than that of SO₄.     

Delayed consequences of extremely low‐frequency magnetic fields and the influence of adverse environmental conditions on roach Rutilus rutilus embryos

This study presents data collected over a 6 year period on the effects of extremely low‐frequency magnetic fields (MFs) (1·4–1·6 µT, 500 Hz and 1·4–1·6 µT, 72·5 Hz) and MFs in combination with other environmental stressors (elevated temperature, 0·01 mg l^?1 trichlorfon, 0·01 mg l^?1 copper sulphate pentahydrate) on roach Rutilus rutilus embryos. Effects were studied during different stages of early development. Rutilus rutilus were raised in ponds for 4 months after exposure to MFs. The mass, standard length (L_S) and morphological characteristics of underyearlings which were exposed as embryos were recorded. An increase in embryo mortality and a decrease in L_S and mass indices in underyearlings were noted after they had been exposed to a combination of MFs and different adverse environmental factors. In addition, exposure to MFs led to changes in the total number of vertebrae and the number of seismosensory system openings in the mandibular bones of underyearlings. MFs of different frequency caused both increases (500 Hz) and decreases (72·5 Hz) in morphological diversity. The stressors used in this study, however, did not increase the fluctuating asymmetry of bilateral morphological characteristics. The possible microevolutionary effects of exposure to MFs alone and in combination with other adverse environmental factors upon natural fish populations are discussed.     

LIGHT AND TEMPERATURE AS FACTORS REGULATING SEASONAL GROWTH AND DISTRIBUTION OF ULOTHRIX ZONATA (ULVOPHYCEAE)1

Ulothrix zonata (Weber and Mohr) Kütz. is an unbranched filamentous green alga found in rocky littoral areas of many northern lakes. Field observations of its seasonal and spatial distribution indicated that it should have a low temperature and a high irradiance optimum for net photosynthesis, and at temperatures above 10°C it should show an increasingly unfavorable energy balance. Measurements of net photosynthesis and respiration were made at 56 combinations of light and temperature. Optimum conditions were 5°C and 1100 μE·m^?2·s^?1 at which net photosynthesis was 16.8 mg O₂·g^?1·h^?1. As temperature increased above 5° C optimum irradiance decreased to 125 μE·m^?2·s^?1 at 30°C. Respiration rates increased with both temperature and prior irradiance. Light-enhanced respiration rates were significantly greater than dark respiration rates following irradiance exposures of 125 μE·m^?2·s^?1 or greater. Polynomials were fitted to the data to generate response surfaces. Polynomial equations represent statistical models which can accurately predict photosynthesis and respiration for inclusion in ecosystem models.     

EFFECT OF TEMPERATURE AND IRRADIANCE ON GROWTH OF HAEMATOCOCCUS PLUVIALIS (CHLOROPHYCEAE)1

The growth characteristics of Haematococcus pluvialis Flotow were determined in batch culture. Optimal temperature for growth of the alga was between 25° and 28°C, at which the specific growth rate was 0.054 h^?1. At higher temperatures, no cell division was observed, and cell diameter increased from 5 to 25 μm. The saturated irradiance for growth of the alga was 90 μmol quanta · m^?2·s^?1; under higher irradiances (e.g. 400 μmol quanta·m^?2·s^?1) astaxanthin accumulation was induced. Growth rate, cell cycle, and astaxanthin accumulation were significantly affected by growth conditions. Careful attention should be given to the use of optimal growth conditions when studying these processes.     

THE INABILITY OF THE TEXAS “BROWN TIDE” ALGA TO USE NITRATE AND THE ROLE OF NITROGEN IN THE INITIATION OF A PERSISTENT BLOOM OF THIS ORGANISM1

A planktonic alga similar in general morphology and pigments to Aureococcus anophagefferens Hargraves and Sieburth has caused persistent and ecologically damaging blooms along the south Texas coast. Experiments using 100 μM NO₃^?, NO₂^?, and NH₄⁺ demonstrated that the alga could not use NO₃^? for growth but could use NO₂^? and NH₄⁺. Doubling iron or trace metal concentrations did not permit growth on NO₃^?. Chemical composition data for cultures grown in excess NO₃^? or NH₄⁺, respectively, were as follows: N·cell^?1 (0.88 vs. 1.3 pg), C:N ratio (25:1 vs. 6.4:1), C:chlorophyll a (chl a) (560:1 vs. 44:1), and chl a·cell^?1 (0.033 vs. 0.16 pg). These data imply that cells supplied with NO₃^? were N-starved. Culture addition of 10 mM final concentration chlorate (a nitrate analog) did not affect the Texas isolate while NO₃^? utilizing A. anophagefferens was lysed, suggesting that the NO₃^? reductase of the Texas isolate is nonfunctional. Rates of primary productivity determined during a dense bloom indicated that light-saturated growth rates were ca. 0.45 d^?1, which is similar to maximum rates determined in laboratory experiments (0.58 d^?1± 0.16). However, chemical composition data were consistent with the growth rate of these cells being limited by N availability (C:N 28, C:chl a 176, chl a·cell^?1 0.019). Calculations based on a mass balance for nitrogen suggest that the bloom was triggered by an input of ca. 69 μM NH₄⁺ that resulted from an extensive die-off of benthos and fish.     

Plant growth regulators affect biosynthesis and accumulation profile of isoflavone phytoestrogens in high-productive in vitro cultures of Genista tinctoria

The influence of plant growth regulators on biomass growth and the accumulation of medicinally-relevant isoflavone phytoestrogens, derivatives of genistein and daidzein (8 compounds including aglycones, glucosides and glucoside esters) in callus cultures of Genista tinctoria (Fabaceae) was examined. The experiments included 10 auxins [2,4-dichlorophenoxyacetic acid (2,4-D), p-chlorophenoxyacetic acid, indole-3-acetic acid, indole-3-butyric acid, indole-3-propionic acid, 1-naphthaleneacetic acid, β-naphthoxyacetic acid, picloram, 2,3,5-triiodobenzoic acid (TIBA), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T)] and 7 cytokinins [6-benzylaminopurine, forchlorfenuron, 1,3-diphenylurea, 2-isopentenyladenine, kinetin (KIN), thidiazuron, zeatin] applied at 0.5 and 5.0 mg l^?1, jointly with 5.0 or 0.5 mg l^?1 KIN or 2,4-D (for auxins and cytokinins, respectively—36 phytohormone combinations in total). Statistical analysis of the relationships between callus growth [expressed as growth index (Gi)] and the accumulation of isoflavones showed positive correlation in the cytokinin group (r_xy values from 0.13 to 0.61) and negative correlation within auxins (r_xy values from ?0.31 to ?0.39). Among the cytokinins tested, the highest isoflavone content (6,436.26 mg/100 g dry weight) and the fastest biomass growth (Gi = 892.46 %) were obtained for 0.5 mg l^?1 KIN used jointly with 5.0 mg l^?1 2,4-D. In the group of auxins, the combination of 0.5 mg l^?1 TIBA and 5.0 mg l^?1 KIN provided the fastest culture growth (Gi = 983.07 %) and the isoflavone concentration of 10,474.23 mg/100 g dry weight, which is so far the highest amount of these metabolites achieved in callus cultures of higher plants.     

Protein recovery by continuous flotation

Summary Bovine serum albumin (BSA) was recovered from aqueous solutions by foam flotation. The protein concentrations in foam liquid C _S, in feed C _Pand in residue C _Rwere determined. The protein enrichment C _S/C_Pand the separation C _S/C_Ras well as the protein fraction in the foam liquid % BSA and foam liquid volume flow were determined as functions of the medium properties, operational conditions, and equipment parameters as well as concentrations of solid particles. At low protein concentrations in feed (e.g., C _P=40 mg · l^-1), and at 40° C, high performance was attained (C _X=2,000 mg · l^-1, C _R=4.4 mg · l^-1, C _S/C_P=50, C _S/C_R=450, 90% BSA. Continuous foam flotation is an efficient procedure for the recovery of low concentrations of proteins from liquid cultures.Abbreviations BSA bovine serum albumine - C _P protein concentration in feed (mg · l^-1) - C _R protein concentration in residue (mg · l^-1) - C _S protein concentration in foam liquid (mg · l^-1) - C _S/C_R protein separation (-) - C _S/C_P protein enrichment (-) - V _P feed rate (ml · min^-1) - V _R residue flow rate (ml · min^-1) - V _S foam liquid volume flow (ml · min^-1) - N number of theoretical stages in an ideal cascade (-) - temperature (° C) - mean residence time (min)     

OSCILLATORIA (TRICHODESMIUM) THIEBAUTII (CYANOPHYTA) IN THE CENTRAL NORTH ATLANTIC OCEAN12

Physiological rate measurements were made with Oscillatoria thiebautii (Gom.) Geitler in the subtropical north Atlantic Ocean between Spain and Bermuda during May and June of 1975. The near surface C:N fixation ratios averaged 6.5, and the cellular composition ratio was 6.2, suggesting that N₂ fixation is the major path of nitrogenous nutrition for this alga. Compared to other oceanic phytoplankters, it has a low affinity for orthophosphate at oceanic concentrations (k_s= 9.0); however, it has a high potential for utilizing phosphomonoesters (170–300 ng atoms P ·μg chl a^?1· h^?1). Maximal photosynthesis occurred at 450–700 μ Einstein · m^?2· s^?1, and was inhibited by full sunlight. Calculated cell division rates (ca. 180 days) suggest that relative to other phytoplankters in this oceanic region, O. thiebautii must be subjected to negligible grazing pressure. No major differences in C, N, chl a or ATP were observed between the tuft (fusiform) and puff (spherical) colonies. ATP concentrations relative to other cellular constituents varied greatly between colonies, suggesting a general inter-colony physiological variability in the open Atlantic. With increasing depth in the euphotic zone, there was no evidence for chromatic adaption. The observations that O. thiebautii represents only a small fraction of total phytoplankton biomass and that its growth rate is 10–100 times slower than that of the other indigenous phytoplankton, strongly suggest that N₂ fixation by this alga is a virtually insignificant component of the nitrogenous nutrition for the phytoplankton of the North Atlantic central gyre in late Spring.     

Use chemometric techniques in the optimization of a specific bioassay for betalactams in milk

Aims: A microbiological bioassay using Geoacillus stearothermophilus was optimized to detect betalactams at concentrations near to the Maximum Residue Limits (MRLs), with low cross‐specificity for tetracycline. Methods and Results: A factorial design (3 × 4) was used to evaluate the effects of concentration of spores (2·0 × 10⁶, 4·0 × 10⁶ and 8·0 × 10⁶ spores ml^?1) and incubation time (3·0, 3·5, 4·0 and 4·5 h) on the response of the bioassay. Then, desirability function to raise the detection capabilities (CC_β) of tetracyclines and increase sensitivity to betalactams was implemented. Significant effects of Log[S] and incubation time [It] on the CC_β of betalactams and tetracyclines were observed. Finally, high value of global desirability (D = 0·853), adequate betalactams CC_β (3·8 μg l^?1 of penicillin ‘G’, 27 μg l^?1 of oxacillin, 8·1 μg l^?1 of ampicillin, 48 μg l^?1 of cloxacillin) and high tetracyclines CC_β (5260 μg l^?1 chlortetracycline, 1550 μg l^?1 of oxytetracycline, 1070 μg l^?1 of tetracycline) were calculated. Conclusions: The application of chemometric tools allows the optimization of a bioassay that detects betalactam residues in milk. The more robust conditions have been achieved in Log[S] = 6·30 and [It] = 4·20 h. Significance and Impact of the Study: The logistic regression model and the desirability function are adequate chemometric techniques to improve the properties of the methods, because it is possible to increase sensitivity and decrease cross‐specificity simultaneously.     

Effects of ascorbic acid,alpha-tocopherol,and glutathione on microspore embryogenesis in Brassica napus L.

The impact of culture conditions and addition of antioxidants to media on microspore embryogenesis in rapeseed (Brassica napus cv. ‘PF704’) was investigated. Different concentrations of ascorbic acid (0, 5, 10, 20, 50, 100, and 200 mg l^?1) and alpha (α)-tocopherol (0, 5, 10, 20, 50, 100, and 200 mg l^?1) were evaluated along with two temperature pretreatments (18 d at 30°C; 2 d at 32.5°C followed by 16 d at 30°C). In addition, combinations of reduced glutathione (0, 10, 50, and 100 mg l^?1) and ascorbic acid (5 and 10 mg l^?1) were tested. Microspore embryogenesis was significantly enhanced using 10 mg l^?1 ascorbic acid (334 embryos per Petri dish) compared with untreated cultures (184 embryos per Petri dish) at 30°C. α-Tocopherol (5 and 10 mg l^?1) enhanced (312 and 314 embryos per Petri dish, respectively) microspore embryogenesis relative to untreated cultures (213 embryos per Petri dish) at 30°C, although there were no significant differences among cultures treated with 5–50 mg l^?1 α-tocopherol. When 50 mg l^?1 α-tocopherol was combined with 5 or 10 mg l^?1 ascorbic acid, embryogenesis was significantly enhanced (308 and 328 embryos per Petri dish, respectively) relative to other ascorbic acid levels. Moreover, 10 mg l^?1 of reduced glutathione and 5 mg l^?l ascorbic acid enhanced microspore embryogenesis (335 embryos per Petri dish) compared to cultures without reduced glutathione (275 embryos per Petri dish). Microspore embryogenesis could be improved by adding ascorbic acid, α-tocopherol, and reduced glutathione when the appropriate combination and temperature pretreatment were selected.     

SEASONAL EFFECT ON THE RELATIONSHIP BETWEEN THE PHOTOSYNTHETIC CAPACITY OF INTERTIDAL MICROPHYTOBENTHOS AND TEMPERATURE1

The response of the photosynthetic capacity (P_max) of microphytobenthos to short-term variations of temperature (in the range 5–35° C) was assessed on a seasonal basis. The relationship is described mathematically, and relevant physiological parameters are identified: P_MAX, the maximum value of P_max achieved at T_opl, the optimum temperature. Estimated values of T_opt do not change significantly throughout the year and remain close to 25° C. It is thus concluded that T_opt is not influenced by seasonal variations in the daily range of mud surface temperature. Identical conclusions hold for T_max (ca. 38° C), the thermal threshold beyond which no photosynthesis occurs. Conversely, P_MA estimates exhibit substantial variability: P_MAX (mean ± root mean square error) is highest in April (11.18 ± 0.42 [μg C · [μg Chl a]^?1· h^?1) during the beginning of the annual increase in temperature, photoperiod, and maximum irradiance and is lowest in December (3.04 ± 0.16 μg C · [μg Chl a]^?1· h^?l). From an ecological point of view, the short-term and seasonal variations of P_MAX suggest that the microphytobenthic community takes advantage of the abiotic spring environmental conditions, allowing the onset of the bloom. Nevertheless, no “acclimation strategy” (i.e. shifts in T_opt and T_max that prevent temperature inhibition in summer or improve photosynthetic rates in winter) is apparent from our results.     

Ventilatory and behavioural responses of the marbled sole Pseudopleuronectes yokohamae to progressive hypoxia

This study identified ventilatory and behavioural responses in the marbled sole Pseudopleuronectes yokohamae under experimentally induced progressive decreases in dissolved oxygen (DO) levels. Ventilation frequency showed an increase with decreasing DO levels from normoxia to 2·75 mg O₂ l^?1, followed by a decrease in ventilation frequency at decreased DO levels from 2·00 to 0·75 mg O₂ l^?1. At DO levels below 2·00 mg l^?1, behaviours at the bottom were suppressed, whereas avoidance behaviours increased. A decrease in avoidance behaviours was observed from 1·00 to 0·75 mg O₂ l^?1. Upside‐down reversal and incapacitation at DO levels of 1·00–0·75 mg O₂ l^?1 suggested that sublethal effects on P. yokohamae were induced. The responses observed before the sublethal DO level could be interpreted as an effort to maintain oxygen uptake, reduce routine activities and facilitate avoidance. The observed DO level thresholds that induce behavioural responses, in addition to sublethal effects, indicate hypoxia‐tolerance that is important for understanding the effects of hypoxia on coastal ecosystems.     

Geothermal diatoms: a comparative study of floras in hot spring systems of Iceland,New Zealand,and Kenya

Diatom floras were examined from geothermal environments in three contrasting tectonic settings. These included subduction-related acid and alkaline springs in New Zealand; alkaline springs along a divergent plate boundary on Iceland; and alkaline springs in the Kenya Rift. These shallow (<1 cm) aquatic environments vary considerably (e.g., temperature: 21.3–99°C; pH: 2.1–9.65; 56.41–643 mg l⁻¹ SiO₂). Diatoms form an important component of geothermal floras at temperatures of <45°C. The floras from New Zealand are distinguished by the common occurrence of Pinnularia. Icelandic springs have a variety of Fragilariaceae. Navicula and Anomoeoneis are most common in the Kenyan springs. Statistical analyses suggest that the diatoms cluster into seven major groups. The most common taxa include: Achnanthidium exiguum v. heterovalvum (Kras.) Czarn., Anomoeoneis sphaerophora (Ehrenb.) Pfitz, Brachysira brebissonii f. thermalis Grun., Caloneis bacillum (Grun.) Cl., Craticula cuspidata (Kütz.) Mann, Diadesmis confervacea Kütz., Epithemia argus (Ehrenb.) Kütz., Frustulia rhomboides (Ehrenb.) DeT., Hantzschia amphioxys (Ehrenb.) Grun., Navicula tenelloides Hust., Nitzschia amphibia Grun., Nitzschia inconspicua Grun., Nitzschia invisitata Hust., Nitzschia frustulum (Kütz.) Grun., Nitzschia sigma (Kütz.) W, Smith., Pinnularia chapmaniana Fog., Pinnularia appendiculata (Ag.) Cl., Pinnularia molaris (Grun.) Cl., Pinnularia acoricola Hust., Rhopalodia gibberula (Ehrenb.) O. Müll., Staurosira construens v. venter (Ehrenb.) Ham., Staurosira elliptica (Schum.) Will. & Round, and Staurosirella pinnata (Ehrenb.) Will. & Round. Canonical correspondence analysis shows clear correlations between species, alkalinity, pH, and conductivity, with less strong correlations for silica and temperature. Other factors include substrate type, current velocity, and light conditions. The preservation potential of host deposits varies considerably, being lowest for springs on clastic deltas and highest where travertine or sinter is accumulating. Handling editor: J. Padisak     

EFFECT OF THE SINKING RATE OF TWO DIATOMS (THALASSIOSIRA SPP.) ON UPTAKE FROM LOW CONCENTRATIONS OF PHOSPHATE1

The effect of the sinking rate, or rate of medium flow (φ) on the rate of phosphate incorporation (V) by the planktonic diatoms Thalassiosira fluviatilis Hust. and T. pseudonana Hasle & Heimdal in batch and chemostat cultures was determined by passing medium at defined flow rates (0.5–25.0 mm·min^?1) over algae on membrane filters. At concentrations from 1 to 100 μg phosphorus·l^?1 V, increases with increasing velocity of flow, approaching a maximum value (V_m) as described by the empirical relationship: where K_φ is the sinking rate value when V = 1/2 V_m+ V_o and V_o is the uptake at 0 rate of flow. By comparing uptake at controlled flow with uptake in a vigorously stirred medium, the phosphate concentration in the cell boundary layer can be determined. The sinking rate that reduces the phosphate concentration in the boundary layer to half of nominal concentration in the medium is much lower for the larger T. fluviatilis than for T. pseudonana. For both diatoms, it is inversely related to the nominal concentration.     

THE EFFECT OF NUTRIENT LIMITATION AND ITS INTERACTION WITH LIGHT UPON THE PRODUCTS OF PHOTOSYNTHESIS IN MERISMOPEDIA TENUISSIMA (CYANOPHYCEAE)1

The metabolic fate of photosynthetically-fixed CO₂ was determined by labeling samples of Merismopedia tenuissima Lemmerman for 30 min with NaH¹⁴CO₃ and analyzing its incorporation into low molecular weight compounds, polysaccharide and protein. In N- and P-sufficient cultures, relative incorporation into protein increased as the irradiance used during the labeling period was decreased to 20 μE · m^-2 s^-1. This pattern was found for cells grown at irradiances of either 20 or 180 μE · m^-2· s^-1, although incorporation into protein was greater in cultures grown at the higher irradiance. In N-limited continuous cultures, relative incorporation into protein was low, independent of growth rate, and the same for samples tested at 20 or 180 μE · m^-2· s^-1 irradiance. In contrast, ¹⁴C incorporation into protein by P-limited cultures increased as growth rate increased, and at relative growth rates greater than 0.25, the incorporation was greater at 20 than at 180 μE · m^-2· s^-1. However, the total RNA content and maximum photosynthetic rate of the cultures was the same at all growth rates tested. The interaction between nutrient concentration and light intensity was studied by growing-limited continuous cultures at the same dilution rate, but different irradiances. Relative incorporation into protein was highest in cultures grown at 20 μE · m^-2· s^-1, in which the relative growth rate was 0.4. These results suggest that photosynthetic carbon metabolism may respond to relative growth rate μ/μ_max rather than to growth rate directly.     

THE THERMOPHILIC CYANOPHYTES OF ICELAND AND THE UPPER TEMPERATURE LIMIT1

The upper temperature limit for the growth of chlorophyllous cyanophytes was estimated at about 63–64 C by observations of field populations and by the growth of culture isolates at that temperature. Several isolates of Mastigocladus laminosus were made from different Iceland springs and temperatures. A few continued to grow at 62 C and one at 64 C. M. laminosus was the cyanophyte in Iceland tolerant of the highest temperature. The several species or races of narrow-cell-form Synechococcus, which almost exclusively inhabit North American and other springs above 62 C and up to about 74 C, were not found in Iceland. According to my observations and those of others, a maximum of 6–8 cyanophyte species occur in waters over 45 C in all of Iceland. Many times this number has been reported from continental regions of great thermal activity. The low number of species and the absence of particular thermophilic cyanophytes may result from inadequate dispersal from other thermal areas since the end of the last glaciation. Environmental inadequacies (e.g., winter darkness, or minimum-maximum concentrations of nutrients or other solutes) may also be factors in the absence of certain species.     

Plant regeneration of the arsenic hyperaccumulator <Emphasis Type="Italic">Pteris vittata</Emphasis> L<Emphasis Type="Italic">.</Emphasis> from spores and identification of its tolerance and accumulation of arsenic and copper

An in vitro plant regeneration system was established from the spores of Pteris vittata and identification of its tolerance, and accumulation of gametophytes and callous, to arsenic (As) and copper (Cu) was investigated. The highest frequency (100%) of callus formation was achieved from gametophyte explants treated with 0.5 mg l^?1 6-benzylaminopurine (6-BA) + 0.5 mg l^?1 gibberellin acid (GA). Furthermore, sporophytes were differentiated from the callus tissue derived from gametophyte explants on MS medium supplemented with 0.5 mg l^?1 6-BA, 0.5–1.0 mg l^?1 GA and additional 300 mg l^?1 lactalbumin hydrolysate (LH) for 4 weeks. The optimum combination of ½ MS + 1.0 mg l^?1 GA + 0.5 mg l^?1 6-BA + 300 mg l^?1 LH promoted sporophyte formation on 75 ± 10% of the callus. Every callus derived from gametophyte explants could achieve 3–4 sporophytes. The in vitro growth of gametophyte and callus was accelerated in the medium containing Na₃AsO₄ lower than 0.5 mM, but this growth was inhibited with 2 mM Na₃AsO₄. And with the increase of Na₃AsO₄ in the culture medium from 0 to 2 mM, the As accumulation in gametophytes and callus increased and achieved a level of 763.3 and 315.4 mg kg^?1, respectively. Gametophytes and calluses transplanted to culture medium, supplemented with different concentrations of CuSO₄, are similar to those in Na₃AsO₄, and the Cu accumulation in gametophytes could achieve 7,940 mg kg^?1 when gametophytes were subcultured in medium containing 3 mM CuSO₄. These results suggested that the high efficiency propagation system could be a useful and rapid means to identify other heavy metal tolerance and accumulation. Further, the regeneration ability of callus made it possible for genetic transformation of this fern.