Adaptation to warming but not cooling at slow rates of stimulus change in thermal threshold measurements

The relationship between thermal detection threshold and rate of temperature change of the thermal stimulus when slow (<1 degrees C s(-1)) rates of change are employed was investigated. Using both the reaction time (RT) inclusive Method of Limits and RT exclusive Method of Levels healthy volunteers had warming (WDT) and cooling detection thresholds (CDT) measured at four different rates of temperature change (0.3, 0.5, 0.7 and 1.0 degrees C s(-1)) from the thenar and/or mental regions using a contact thermode. With the Method of Limits, CDT increased linearly with rate of temperature change suggesting increments were due to RT artefacts. This was further supported by threshold assessment with the Method of Levels which showed CDT were unaffected by the rate of change in the RT exclusive method (P > 0.1). In contrast, WDT did not increase linearly with rate of stimulus temperature change when the Method of Limits was used and threshold assessment with the Method of Levels showed WDT assessed using a 0.3 degrees C s(-1) ramp rate were significantly higher than those measured with a 1 degrees C s(-1) rate of change (P < 0.05). This study indicates that adaptation to a warming stimulus can occur at faster rates of stimulus change than previously anticipated and identifies differences in warming and cooling pathways in sensitivity to adaptation.     

Measurement of cooling and warming rates in vitrification‐based plant cryopreservation protocols

Cryopreservation protocols include the use of additives and pretreatments aimed to reduce the probability of ice nucleation at all temperatures, mainly through micro‐viscosity increase. Still, there is a risk of ice formation in the temperature region comprised between the equilibrium freezing (T_f) and the glass transition (T_G) temperatures. Consequently, fast cooling and warming, especially in this region, is a must to avoid ice‐derived damage. Vitrification and droplet‐vitrification techniques, frequently used cryopreservation protocols based in fast cooling, were studied, alongside with the corresponding warming procedures. A very fast data acquisition system, able to read very low temperatures, down to that of liquid nitrogen, was employed. Cooling rates, measured between ?20°C and ?120°C, ranged from ca. 5°C s^?1 to 400°C s^?1, while warming rates spanned from ca. 2°C s^?1 to 280°C s^?1, for the different protocols and conditions studied. A wider measuring window (0°C to ?150°C) produced lower rates for all cases. The cooling and warming rates were also related to the survival observed after the different procedures. Those protocols with the faster rates yielded the highest survival percentages. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1177–1184, 2014     

Mediterranean warming triggers seagrass (Posidonia oceanica) shoot mortality

Rapid warming of the Mediterranean Sea threatens marine biodiversity, particularly key ecosystems already stressed by other impacts such as Posidonia oceanica meadows. A 6‐year monitoring of seawater temperature and annual P. oceanica shoot demography at Cabrera Archipelago National Park (Balearic Islands, Western Mediterranean) allowed us to determine if warming influenced shoot mortality and recruitment rates of seagrasses growing in relative pristine environments. The average annual maximum temperature for 2002–2006 was 1 °C above temperatures recorded in 1988–1999 (26.6 °C), two heat waves impacted the region (with seawater warming up to 28.83 °C in 2003 and to 28.54 °C in 2006) and the cumulative temperature anomaly, above the 1988–1999 mean annual maximum temperature, during the growing season (i.e. degree‐days) ranged between 0 °C in 2002 and 70 °C in 2003. Median annual P. oceanica shoot mortality rates varied from 0.067 year^?1 in 2002 to 0.123 year^?1 in 2003, and exceeded recruitment rates in all stations and years except in shallow stations for year 2004. Interannual fluctuations in shoot recruitment were independent of seawater warming (P>0.05). P. oceanica meadows experienced a decline throughout the study period at an average rate of ?0.050±0.020 year^?1. Interannual variability in P. oceanica shoot mortality was coupled (R²>0.40) to seawater warming variability and increasing water depth: shoot mortality rates increased by 0.022 year^?1 (i.e. an additional 2% year^?1) for each additional degree of annual maximum temperature and by 0.001 year^?1 (i.e. 0.1% year^?1) for each accumulated degree water temperature remained above 26.6 °C during the growing season. These results demonstrate that P. oceanica meadows are highly vulnerable to warming, which can induce steep declines in shoot abundance as well indicating that climate change poses a significant threat to this important habitat.     

The relative impacts of daytime and night-time warming on photosynthetic capacity in Populus deltoides

In order to investigate the relative impacts of increases in day and night temperature on tree carbon relations, we measured night‐time respiration and daytime photosynthesis of leaves in canopies of 4‐m‐tall cottonwood (Populus deltoides Bartr. ex Marsh) trees experiencing three daytime temperatures (25, 28 or 31 °C) and either (i) a constant nocturnal temperature of 20 °C or (ii) increasing nocturnal temperatures (15, 20 or 25 °C). In the first (day warming only) experiment, rates of night‐time leaf dark respiration (R_dark) remained constant and leaves displayed a modest increase (11%) in light‐saturated photosynthetic capacity (A_max) during the day (1000–1300 h) over the 6 °C range. In the second (dual night and day warming) experiment, R_dark increased by 77% when nocturnal temperatures were increased from 15 °C (0·36 µmol m^?2 s^?1) to 25 °C (0·64 µmol m^?2 s^?1). A_max responded positively to the additional nocturnal warming, and increased by 38 and 64% in the 20/28 and 25/31 °C treatments, respectively, compared with the 15/25 °C treatment. These increases in photosynthetic capacity were associated with strong increases in the maximum carboxylation rate of rubisco (V_cmax) and ribulose‐1,5‐bisphosphate (RuBP) regeneration capacity mediated by maximum electron transport rate (J_max). Leaf soluble sugar and starch concentration, measured at sunrise, declined significantly as nocturnal temperature increased. The nocturnal temperature manipulation resulted in a significant inverse relationship between A_max and pre‐dawn leaf carbohydrate status. Independent measurements of the temperature response of photosynthesis indicated that the optimum temperature (T_opt) acclimated fully to the 6 °C range of temperature imposed in the daytime warming. Our findings are consistent with the hypothesis that elevated night‐time temperature increases photosynthetic capacity during the following light period through a respiratory‐driven reduction in leaf carbohydrate concentration. These responses indicate that predicted increases in night‐time minimum temperatures may have a significant influence on net plant carbon uptake.     

TEMPERATURE AND IRRADIANCE EFFECTS ON GROWTH OF PITHOPHORA OEDOGONIA (CHLOROPHYCEAE) AND SPIROGYRA SP. (CHAROPHYCEAE)1

Growth responses of Pithophora oedogonia (Mont.) Wittr. and Spirogyra sp. to nine combinations of temperature (15°, 25°, and 35°C) and photon flux rate (50, 100, and 500 μmol·m^?2·s^?1) were determined using a three-factorial design. Maximum growth rates were measured at 35°C and 500 pmol·m^?2·s^?1 for P. oedogonia (0.247 d^?1) and 25°C and 500 μmol·m^?2·s^?1 for Spirogyra sp. (0.224 d^?1). Growth rates of P. oedogonia were strongly inhibited at 15°C (average decrease= 89%of maximum rate), indicating that this species is warm stenothermal. Growth rates of Spirogyra sp. were only moderately inhibited at 15° and 35°C (average decrease = 36 and 30%, respectively), suggesting that this species is eurythermal over the temperature range employed. Photon flux rate had a greater influence on growth of Spirogyra sp. (31% reduction at 50 pmol·m^?2·s^?1 and 25°C) than it did on growth of P. oedogonia (16% reduction at 50 μmol·m^?2·s^?1 and 35°C). Spirogyra sp. also exhibited much greater adjustments to its content of chlorophyll a (0.22–3.34 μg·mg fwt^?1) than did P. oedogonia (1.35–3.08 μg·mg fwt^?1). The chlorophyll a content of Spirogyra sp. increased in response to both reductions in photon flux rate and high temperatures (35°C). Observed species differences are discussed with respect to in situ patterns of seasonal abundance in Surrey Lake, Indiana, the effect of algal mat anatomy on the internal light environment, and the process of acclimation to changes in temperature and irradiance conditions.     

Temperature × light interaction and tolerance of high water temperature in the planktonic freshwater flagellates Cryptomonas (Cryptophyceae) and Dinobryon (Chrysophyceae)

Using microcosm experiments, we investigated the interactive effects of temperature and light on specific growth rates of three species each of the phytoplanktonic genera Cryptomonas and Dinobryon. Several species of these genera play important roles in the food web of lakes and seem to be sensitive to high water temperature. We measured growth rates at three to four photon flux densities ranging from 10 to 240 μmol photon · m^?2 · s^?1 and at 4–5 temperatures ranging from 10°C to 28°C. The temperature × light interaction was generally strong, species specific, and also genus specific. Five of the six species studied tolerated 25°C when light availability was high; however, low light reduced tolerance of high temperatures. Growth rates of all six species were unaffected by temperature in the 10°C–15°C range at light levels ≤50 μmol photon · m^?2 · s^?1. At high light, growth rates of Cryptomonas spp. increased with temperature until the temperature optimum was reached and then declined. The Dinobryon species were less sensitive than Cryptomonas spp. to photon flux densities of 40 μmol photon · m^?2 · s^?1 and 200 μmol photon · m^?2 · s^?1 over the entire temperature range but did not grow under a combination of very low light (10 μmol photon · m^?2 · s^?1) and high temperature (≥20°C). Among the three Cryptomonas species, cell volume declined with temperature and the maximum temperature tolerated was negatively related to cell size. Since Cryptomonas is important food for microzooplankton, these trends may affect the pelagic carbon flow if lake warming continues.     

Respiration of thermogenic inflorescences of skunk cabbage Symplocarpus renifolius in heliox

The respiration rate of the thermogenic inflorescences of Japanese skunk cabbage Symplocarpus renifolius can reach 300 nmol s^?1 g^?1, which is sufficient to raise spadix temperature (T_s) up to 15 ° C above ambient air temperature (T_a). Respiration rate is inversely related to T_a, such that the T_s achieves a degree of independence from T_a, an effect known as temperature regulation. Here, we measure oxygen consumption rate (?o ₂) in air (21% O₂ in mainly N₂) and in heliox (21% O₂ in He) to investigate the diffusive conductance of the network of gas‐filled spaces and the thermoregulatory response. When T_s was clamped at 15 ° C, the temperature that produces maximal ?o ₂ in this species, exposure to high diffusivity heliox increased mean ?o ₂ significantly from 137 ± 17 to 202 ± 43 nmol s^?1 g^?1 FW, indicating that respiration in air is normally limited by diffusion in the gas phase and some mitochondria are unsaturated. When T_a was clamped at 15 ° C and T_s was allowed to vary, exposure to heliox reduced T_s 1 ° C and increased ?o ₂ significantly from 116 ± 10 to 137 ± 19 nmol s^?1 g^?1, indicating that enhanced heat loss by conduction and convection can elicit the thermoregulatory response.     

Temperature response of photosynthetic light‐ and carbon‐use characteristics in the red seaweed Gracilariopsis lemaneiformis (Gracilariales,Rhodophyta)

The red seaweed Gracilariopsis is an important crop extensively cultivated in China for high‐quality raw agar. In the cultivation site at Nanao Island, Shantou, China, G. lemaneiformis experiences high variability in environmental conditions like seawater temperature. In this study, G. lemaneiformis was cultured at 12, 19, or 26°C for 3 weeks, to examine its photosynthetic acclimation to changing temperature. Growth rates were highest in G. lemaneiformis thalli grown at 19°C, and were reduced with either decreased or increased temperature. The irradiance‐saturated rate of photosynthesis (P_max) decreased with decreasing temperature, but increased significantly with prolonged cultivation at lower temperatures, indicating the potential for photosynthesis acclimation to lower temperature. Moreover, P_max increased with increasing temperature (~30 μmol O₂ · g^?1FW · h^?1 at 12°C to 70 μmol O₂ · g^?1FW · h^?1 at 26°C). The irradiance compensation point for photosynthesis (I_c) decreased significantly with increasing temperature (28 μmol photons · m^?2 · s^?1 at high temperature vs. 38 μmol photons · m^?2 · s^?1 at low temperature). Both the photosynthetic light‐ and carbon‐use efficiencies increased with increasing growth or temperatures (from 12°C to 26°C). The results suggested that the thermal acclimation of photosynthetic performance of G. lemaneiformis would have important ecophysiological implications in sea cultivation for improving photosynthesis at low temperature and maintaining high standing biomass during summer. Ongoing climate change (increasing atmospheric CO₂ and global warming) may enhance biomass production in G. lemaneiformis mariculture through the improved photosynthetic performances in response to increasing temperature.     

PHYSIOLOGICAL RESPONSES OF ANABAENA VARIABILIS (CYANOPHYCEAE) TO INSTANTANEOUS EXPOSURE TO VARIOUS COMBINATIONS OF LIGHT INTENSITY AND TEMPERATURE1

Light intensity and temperature interactions have a complex effect on the physiological process rates of the filamentous bluegreen alga Anabaena variabilis Kütz. The optimum temperature for photosynthesis increased with increasing light intensity from 10°C at 42 μE·m^?2·s^?1 to 35°C at 562 μE·m^?2·s^?1. The light saturation parameter, I_K, increased with increasing temperatures. The maximum photosynthetic rate (2.0 g C·g dry wt.^?1·d^?1) occurred at 35°C and 564 μE·m^?2·s^?1. At 15°C, the maximum rate was 1.25 g C·g dry wt.^?1·d^?1 at 332 μE·m^?2·s^?1. The dark respiration rate increased exponentially with temperature. Under favorable conditions of light intensity and temperature the percent of extracellular release of dissolved organic carbon was less than 5% of the total C fixed. This release increased to nearly 40% under combinations of low light intensity and high temperature. A mathematical model was developed to simulate the interaction of light intensity and temperature on photosynthetic rate. The interactive effects were represented by making the light-saturation parameters a function of temperature.     

Influence of light and temperature on photosynthesis and respiration by Pithophora oedogonia (Mont.) Wittr. (chlorophyceae)

Rates of net photosynthesis and respiration were determined for Pithophora oedogonia (Mont.) Wittr. acclimatized to 56 combinations of light (7–1200 μE m^?2 s^?1) and temperature (5–35°C). Conditions for maximum net photosynthesis were estimated to be 26°C and 970 μE m^?2 s^?1. The rate of net photosyntheses varied considerably with temperature, with the maximum measured value (9.67 mg O₂ h^?1 g dry wt.^?1) occurring at 25°C. Respiration rate increased with temperature and the light received just prior to measurement. The maximum respiration rate (7.05 mg O₂ g^?1 h^?1) occurred at 30°C and 1200 μE m^?2 s^?1. Exposure of Pithophora to light levels of 600 or 1200 μE m^?2 s^?1 prior to determination of the respiration rate resulted in significantly elevated levels of oxygen consumption at temperatures ≥ 15°C. The relationship between light, temperature and photosynthesis and respiration were summarized as three-dimensional response surfaces.     

EFFECTS OF LIGHT AND TEMPERATURE ON NET PHOTOSYNTHESIS AND DARK RESPIRATION OF GAMETOPHYTES AND EMBRYONIC SPOROPHYTES OF MACROCYSTIS PYRIFERA1

The rates of net photosynthesis as a function of irradiance and temperature were determined for gametophytes and embryonic sporophytes of the kelp, Macrocystis pyrifera (L.) C. Ag. Gametophytes exhibited higher net photosynthetic rates based on oxygen and pH measurements than their derived embryonic sporophytes, but reached light saturation at comparable irradiance levels. The net photosynthesis of gametophytes reached a maximum of 66.4 mg O₂ g dry wt^?1 h^?1 (86.5 mg CO₂ g dry wt^?1 h^?1), a value approximately seven times the rate reported previously for the adult sporophyte blades. Gametophytes were light saturated at 70 μE m^?2 s^?1 and exhibited a significant decline in photosynthetic performance at irradiances 140 μE m^?1 s^?1. Embryonic sporophytes revealed a maximum photosynthetic capacity of 20.6 mg O₂ g dry wt^?1 h^?1 (25.3 mg CO₂ g dry wt^?1 h^?1), a rate about twice that reported for adult sporophyte blades. Embryonic sporophytes also became light saturated at 70 μE m^?2 s^?1, but unlike their parental gametophytes, failed to exhibit lesser photosynthetic rates at the highest irradiance levels studied; light compensation occurred at 2.8 μE m^?2 s^?1. Light-saturated net photosynthetic rates of gametophytes and embryonic sporophytes varied significantly with temperature. Gametophytes exhibited maximal photosynthesis at 15° to 20° C, whereas embryonic sporophytes maintained comparable rates between 10° and 20° C. Both gametophytes and embryonic sporophytes declined in photosynthetic capacity at 30° C. Dark respiration of gametophytes was uniform from 10° to 25° C, but increased six-fold at 30° C; the rates for embryonic sporophytes were comparable over the entire range of temperatures examined. The broader light and temperature tolerances of the embryonic sporophytes suggest that this stage in the life history of M. pyrifera is well suited for the subtidal benthic environment and for the conditions in the upper levels of the water column.     

SHORT‐TERM EFFECT OF TEMPERATURE ON THE PHOTOKINETICS OF MICROALGAE FROM THE SURFACE LAYERS OF ANTARCTIC PACK ICE1

Microalgae growing within brine channels (85 psu salinity) of the surface ice layers of Antarctic pack ice showed considerable photosynthetic tolerance to the extreme environmental condition. Brine microalgae exposed to temperatures above ?5°C and at irradiances up to 350 μmol photons·m^?2·s^?1 showed no photosynthetic damage or limitations. Photosynthesis was limited (but not photoinhibited) when brine microalgae were exposed to ?10°C, provided the irradiance remained under 50 μmol photons·m^?2·s^?1. The highest level of photosynthetic activity (maximum relative electron transport rate [rETR_max]) in brine microalgae growing within the surface layer of sea ice was at approximately 18 μmol electrons·m^?2·s^?1, which occurred at ?1.8°C. Effective quantum yield of PSII and rETR_max of the halotolerant brine microalgae exhibited a temperature‐dependent pattern, where both parameters were higher at ?1.8°C and lower at ?10°C. Relative ETR_max at temperatures above ?5°C were stable across a wide range of irradiance.     

Helix-coil dynamics of a Z-helix hairpin

The helix–coil transition of a Z-helix hairpin formed from d(C-G)₅T₄(C-G)₅ has been characterized by equilibrium melting and temperature jump experiments in 5M NaClO₄ and 10 mM Na₂HPO₄, pH 7.0. The melting curve can be represented by a simple all-or-none transition with a midpoint at 81.6 ± 0.4°C and an enthalpy change of 287 ± 15 kJ/mole. The temperature jump relaxation can be described by single exponentials at a reasonable accuracy. Amplitudes measured as a function of temperature provide equilibrium parameters consistent with those derived from equilibrium melting curves. The rate constants of Z-helix formation are found in the range from 1800 s^?1 at 70°C to 800 s^?1 at 90°C and are associated with an activation enthalpy of ?(50 ± 10) kJ/mole, whereas the rate constants of helix dissociation are found in the range from 200 s^?1 at 70°C to 4500 s^?1 at 90°C with an activation enthalpy +235 kJ/mole. These parameters are consistent with a requirement of 3–4 base pairs for helix nucleation. Apparently nucleation occurs in the Z-helix conformation, because a separate slow step corresponding to a B to Z transition has not been observed. In summary, the dynamics of the Z-helix–coil transition is very similar to that of previously investigated right-handed double helices.     

Effect of irradiance and temperature on the photosynthesis of a cultivated red alga,Pyropia tenera (= Porphyra tenera), at the southern limit of distribution in Japan

The effect of irradiance and temperature on the photosynthesis of the red alga, Pyropia tenera, was determined for maricultured gametophytes and sporophytes collected from a region that is known as one of the southern limits of its distribution in Japan. Macroscopic gametophytes were examined using both pulse‐amplitude modulated fluorometry and/or dissolved oxygen sensors. A model of the net photosynthesis–irradiance (P‐E) relationship of the gametophytes at 12°C revealed that the net photosynthetic rate quickly increased at irradiances below the estimated saturation irradiance of 46 μmol photons m^?2 s^?1, and the compensation irradiance was 9 μmol photons m^?2 s^?1. Gross photosynthesis and dark respiration for the gametophytes were also determined over a range of temperatures (8–34°C), revealing that the gross photosynthetic rates of 46.3 μmol O₂ mg_chl‐a^?1 min^?1 was highest at 9.3 (95% Bayesian credible interval (BCI): 2.3–14.5)°C, and the dark respiration rate increased at a rate of 0.93 μmol O₂ mg_chl‐a^?1 min^?1°C^?1. The measured dark respiration rates ranged from ?0.06 μmol O₂ mg_chl‐a^?1 min^?1 at 6°C to ?25.2 μmol O₂ mg_chl‐a^?1 min^?1 at 34°C. The highest value of the maximum quantum yield (Fv/Fm) for the gametophytes occurred at 22.4 (BCI: 21.5–23.3) °C and was 0.48 (BCI: 0.475–0.486), although those of the sporophyte occurred at 12.9 (BCI: 7.4–15.1) °C and was 0.52 (BCI: 0.506–0.544). This species may be considered well‐adapted to the current range of seawater temperatures in this region. However, since the gametophytes have such a low temperature requirement, they are most likely close to their tolerable temperatures in the natural environment.     

Temperature effects on oxygen thresholds for hypoxia in marine benthic organisms

The effect of warming on the oxygen requirements and the survival of benthic organisms under hypoxia was tested using a meta‐analysis of published results of experiments evaluating the effects of temperature on the median lethal time and median lethal concentration of benthic macrofauna under hypoxia. The meta‐analysis confirmed that survival times under hypoxia were reduced by on average 74% and that median lethal concentration increased by on average 16% when marine benthic organisms were exposed to warmer temperatures. Warming reduced survival times of marine benthic macrofauna under hypoxia by a median of 3.95±1.67 h °C^?1 and increased the oxygen thresholds for hypoxia‐driven mortality by a median of 1.02±0.15% saturation °C^?1 or 0.07±0.01 mg O₂ L^?1 °C^?1. The corresponding Q₁₀ values averaged 3.01±0.29 for the median survival time and 2.09±0.20 for the median lethal oxygen concentration. Use of these Q₁₀ values predicts that the 4 °C warming expected during the 21st century will lead to survival times 35.6% lower under hypoxia and that the threshold oxygen concentrations for high mortality to occur will increase by, on average, 25.5% if bottom water temperature increased by 4 °C. Hence, ocean warming is expected to increase the vulnerability of benthic macrofauna to reduced oxygen concentrations and expand the area of coastal ecosystems affected by hypoxia.     

COMPARATIVE STUDY ON THE PHOTOSYNTHETIC PROPERTIES OF PRASIOLA (CHLOROPHYCEAE) AND NOSTOC (CYANOPHYCEAE) FROM ANTARCTIC AND NON‐ANTARCTIC SITES1

The terrestrial cyanobacterium Nostoc commune Vaucher ex Bornet et Flahault occurs worldwide, including in Japan and on the Antarctic continent. The terrestrial green alga Prasiola crispa (Lightf.) Kütz. is also distributed in Antarctica. These two species need to acclimate to the severe Antarctic climate including low ambient temperature and desiccation under strong light conditions. To clarify this acclimation process, the physiological characteristics of the photosynthetic systems of these two Antarctic terrestrial organisms were assessed. The relative rate of photosynthetic electron flow in N. commune collected in Japan and in Antarctica reached maxima at 900 and 1,100 μmol photons · m^?2 · s^?1, respectively. The difference seemed to reflect the presence of high amounts of UV‐absorbing substances within the Antarctic cyanobacterium. On the other hand, the optimal temperatures for photosynthesis at the two locations were 30°C–35°C and 20°C–25°C, respectively. This finding suggested a decreased photosynthetic thermotolerance in the Antarctic strain. P. crispa exhibited desiccation tolerance and dehydration‐induced quenching of PSII fluorescence. Re‐reduction of the photooxidized PSI reaction center, P700, was also inhibited at fully dry states. Photosynthetic electron flow in P. crispa reached a maximum at 20°C–25°C and at a light intensity of 700 μmol photons ? m^?2 ? s^?1. Interestingly, the osmolarity of P. crispa cells suggested that photosynthesis is performed using water absorbed in a liquid form rather than water absorbed from the air. Overall, these data suggest that these two species have acclimated to optimally photosynthesize under conditions of the highest light intensity and the highest temperature for their habitat in Antarctica.     

Characterization and comparisons of five N2-fixing Azolla-Anabaena associations,I. Optimization of growth conditions for biomass increase and N content in a controlled environment*

Abstract Biomass increase, C and N content, C₂H₂ reduction, percentage dry weight and chlorophyll a/b ratios were determined for clones of Azolla caroliniana Willd., A. filiculoides Lam., A. mexicana Presl., and A. pinnata R. Br. as a function of nutrient solution, pH, temperature, photoperiod, and light intensity in controlled environment studies. These studies were supplemented by a glasshouse study. Under a 16 h, 26°C day at a light intensity of 200 μmol m^?2 s^?1 and an 8 h, 19° C dark period, there was no significant difference in the growth rates of the individual species on the five nutrient solutions employed. Growth was comparable from pH 5 to pH 8, but decreased at pH 9. Using the same photoperiod and light intensity but constant growth temperatures of 15–40°C, at 5°C intervals, the individual species exhibited maximum growth, nitro-genase (N²ase) activity and N content at either 25° or 30°C. There was no difference in the temperature optima at pH 6 and pH 8. The tolerance of the individual species to elevated temperature was indicated to be A. mexicana> A. pinnata> A. caroliniana> A.filiculoides. At the optimum temperature, growth rates increased with increasing photoperiod at both pH 6 and pH 8 but N₂ase activity was usually highest at a 16 h light period. At photon flux densities of 100, 200, 400 and 600 μmol m^?2 s^?1, during a 16 h light period and optimum growth temperature of the individual species, N₂ase activity was saturated at less than 200 μmol m^?2 s^?1 and growth at 400 μmol m^?2 s^?1.No interacting effects of light and pH were noted for any species, nor were light intensities up to 1700 μmol m^?2 s^?1 detrimental to the growth rate or N content of any species in a 5 week glasshouse study with a natural 14.5 h light period and a constant temperature of 27.5°C. Using the optimum growth temperature, a 16 h light period, and a photon flux density of at least 400 μmol m^?2 s^?1, the Azolla species all doubled their biomass in 2 days or less and contained 5–6% N on a dry weight basis.     

Displacement and Return Movement of Chloroplasts in the Marine Dinophyte Pyrocystis noctiluca. Experiments with Optical Tweezers

Infrared laser traps (optical tweezers) were used to study laser-induced organelle movements in the marine alga Pyrocystis noctiluca (Dinophyta). These cells are highly suitable for optical micromanipulation due to their large size and extensive vacuole. Experiments were done with plastids held by optical tweezers and moved from the nuclear area into the vacuole. The subsequent retraction movement was analysed for speed. The displaced organelles remained connected to their original position by a thin cytoplasmic strand, often less than 1 μm in diameter. When the organelles were released they rapidly returned at an initial rate of 81.7 ± 7.8 μm . s^?1 (overall displacement 50 μm, measured distance 20 μm, 25 °C ± 1 °C, number of cells 22), slowing down with progressive retraction of the connecting strand. The return movement was reduced to 4.2 ± 0.2 μ .s^?1 (n = 10) when the organelles were displaced and held for 1 min. Displacement to a longer distance increased the rate of return movement. A change from a high to a low environmental temperature significantly reduced movement from 94.5 ± 9.0 . s^?1 (30 °C ± 1 °C, n = 22) to 34.5 ± 2.7 μm .s^?1 (5°C ± 1 °C, n = 22). Nocodazole and N-ethylmaleimide (NEM), inhibitors of microtubules and acto-myosin, respectively, did not affect the retraction of the connecting strand, but at high concentrations of NEM it became increasingly difficult to move organelles away from the nuclear area. We suggest that the return movement of organelles within laser-induced artificial strands mainly depends on the viscoelastic properties of the tonoplast. The quantification of these properties by optical tweezers allows determination of reactions of plant cells to temperature changes.     

Ecophysiological adaptations of two Mediterranean red algae in relation to distribution

Effects of irradiance and temperature on the Mediterranean red algae Eupogodon spinellus and Eupogodon planus were tested. Growth of both species was saturated at an irradiance of 10–20?μmol?m^?2?s^?1, which is in accordance with their sublittoral habitat. Eupogodon spinellus and E. planus survived permanently at temperatures between 8 and 30?°C. The temperature optimum for growth was 25?°C with suboptimal growth occurring at (10?)15 and 30?°C in both species. At their collection locality (Corsica), potential monthly growth yields would be highest in summer and in winter would be only about 20% of the maximum. Reproductive requirements could be determined only in E. planus. Gametophytes reproduced both in long and in short days but only at 20?°C. Tetrasporophytes reproduced at 15–20?°C but only in short days. Geographic distribution boundaries are not set by growth or survival limits. However, the reproductive requirements of E. planus did account for its restricted distribution in the Mediterranean and on the Canary Islands.