Secretion of Free Fatty Acids by Prokaryotic and Eukaryotic Algae at Optimal,Supraoptimal, and Suboptimal Growth Temperatures

The paper describes the composition of extracellular free fatty acids (FFAs) and intracellular fatty acids (FAs) in the enrichment cultures of the prokaryotic alga Spirulina platensisand the eukaryotic alga Chlorella vulgarisgrown at optimal, supraoptimal, and suboptimal growth temperatures. With increasing growth temperature, the degree of unsaturation of the intracellular FAs of both algae decreased, while that of the extracellular FFAs of S. platensisincreased. The composition of the extracellular FFAs of C. vulgarispractically did not depend on the growth temperature.     

Photosynthesis and Respiration of Maize Seedlings at Suboptimal Temperatures

Growth of maize seedlings is limited by low temperature andceases at about 10 °C. In a search for processes underlyingthis limitation, rates of photosynthesis and dark respirationwere measured in controlled environments at night/day temperaturesof 5/10, 10/15, and 15/20 °C. CO₂ exchange was measuredon whole seedlings including the root system. Two maize hybrids,which differed in dry matter production at low temperature,were tested from the 4th-leaf until the 8th-leaf stage. Grossphotosynthesis, net photosynthesis, and respiration increasedproportionally with leaf area. The response to low temperaturewas independent of seedling stage. The decline of photosynthesisand respiration with decreasing temperature was similar in bothhybrids. At 10 °C, rates of photosynthesis and respirationwere relatively high indicating a minimum temperature for thoseprocesses much lower than 10 °C. Possible reasons for growthlimitation at 10 °C are discussed.     

Nitrate Signaling and the Two Component High Affinity Uptake System in Arabidopsis

Nitrate transporters are important for nitrogen acquisition by plants and in algae some require two gene products, NRT2 and NAR2, for function. The NRT2 family was already described and the recent identification of a family of the NAR2-type genes in higher plants showed that there was a homologue in Arabidopsis, AtNAR2.1. Using heterologous expression in yeast and oocytes we showed that the two Arabidopsis AtNRT2.1 and AtNAR2.1 proteins interacted to give a functional high affinity nitrate transport system (HATS). The gene knock out mutant atnar2.1-1 is deficient specifically for HATS activity and the resulting growth phenotype on low nitrate concentration is more severe than for the atnrt2.1-1 knock out mutant. Physiological characterisation of the plant N status and gene expression revealed a pattern that was characteristic of severe nitrogen deficiency. Consistent with the down regulation of AtNRT2.1 expression, the atnar2.1-1 plants also displayed the same phenotype as atnrt2.1 mutants in lateral root (LR) response to low nitrate supply. Using atnar2.1-1 plants constitutively expressing the NpNRT2.1 gene, we now show a specific role for AtNAR2.1 in LR response to low nitrate supply. AtNAR2.1 is also involved in the repression of LR initiation in response to high ratios of sucrose to nitrogen in the medium. Therefore the two component system itself is likely to be involved in the signaling pathway integrating nutritional cues for LR architecture regulation. Using a green fluorescent protein-NRT2.1 protein fusion we show the essential role of AtNAR2.1 for the presence of AtNRT2.1 to the plasma membrane.Key Words: high affinity nitrate transport, nitrate transporter, nitrate signalling, root growth     

Nitrogen and Nitrate Accumulation in Species Having Different Relationships Between Nitrate Uptake and Reduction

Species differ in the relationship of nitrate reductase activityto nitrate uptake. In Capsicum annuum different diurnal patternsof leaf nitrate reductase activity and nitrate uptake have beenreported. As a consequence, the relationship of free nitratein the plant to nitrate supplied has a higher level of significancethan has reduced nitrogen to nitrate supplied. In Zea mays ithas been reported that leaf nitrate reductase activity respondsdirectly to nitrate translocation to the leaf and in this speciesthe relationship of greatest significance is reduced nitrogencontent to nitrate supplied. In both species, and also in Cucumis melo, the proportion oftotal plant free nitrate and reduced nitrogen in the roots decreases,and in the stem increases, with increasing nitrate supplied. The accumulation of free nitrate in leaves is accompanied bya quantitatively different relationship between reduced nitrogenand dry weight compared to leaves not accumulating nitrate. Capsicum annuum. L., Cucumis melo L., melon, Zea mays L., maize, sweet corn, nitrate reductase, nitrate uptake     

Isolation of Bacteria Which Can Grow at Both High pH and Low Temperature

Fifty bacterial strains able to grow at pH 10 and 0°C were isolated from soils, and growth characteristics of three selected strains were investigated. Strain 207, which showed the best growth rate of all the isolates at the conditions described above, could grow at a temperature of −5 to 39°C at pH 8.5. The optimum pH for this strain changed from 9.5 at 10°C to 9.0 at 20°C.     

Cultivar Differences in the Performance of Bean Seedlings at Suboptimal Temperatures

Growth analysis of plants raised under controlled environments(10–5, 12, 15, 18 and 20 °C, and 21 h photoperiod)was used to examine whether varietal differences in the minimumgermination temperature of four bean cultivars persist duringgrowth at suboptimal temperatures. A method to estimate theminimum vegetative growth temperature, based on axis relativegrowth rate, was developed. In order to compensate for ontogeneticdrift, the harvests were conducted at the same stage of developmentof the plants. Axis relative growth rates, reduction rates ofthe cotyledons and other growth parameters were calculated inorder to compare the cultivars. Cultivar ‘Marschall’showed better growth potential at 12 °C than the others,‘Pergousa‘ at 15 °C, and ‘Marschall’,‘Olsok’ and ‘Pergousa’ at 18 and 20°C. The effect of temperature on axis RGR was similar for‘Marschall’, ‘Olsok’ and ‘Pergousa’(Q₁₀ = 2·1) and more pronounced than for ‘Processor’(Q₁₀ = 1·3). Although there were significant differencesin the growth parameters among the cultivars within each temperatureused, the differences did not correspond with the differencesduring germination at low temperatures. The minimum vegetativegrowth temperature was close to 10 °C for all the cultivarstested. Phaseolus vulgaris L., beans, suboptimum temperature, growth analysis, minimum germination temperature, minimum vegetative growth temperature     

Spectral Dependence of Photoregulation of Inorganic Nitrogen Metabolism in Chlamydomonas reinhardii

The utilization of NO₃⁻ by green algae growing photoautotrophically under air, which are growth conditions close to their more habitual situations in nature, is associated with the excretion of NO₂⁻ and NH₄⁺ to the culture medium. The entire process is promoted by blue light and depends on photosynthetically active radiation for the required reducing equivalents. The stimulation of NO₃⁻ utilization and of its associated NO₂⁻ and NH₄⁺ excretions saturated at very low quantum fluxes of blue light (15 microequivalents per square meter per second) in Chlamydomonas reinhardii cells sparged with CO₂-free air and irradiated with 50 microequivalents per square meter per second background red light. The wavelength dependence data of this stimulation correlated closely with the in situ photoactivation of nitrate reductase and also with the light induced increase in its biosynthesis and/or assembly.

These results indicate that the photoregulation of inorganic N metabolism in C. reinhardii is mainly due to the blue light modulation of nitrate reductase. Although flavins are the most suitable candidates to act as physiological photoreceptors, the wavelength dependence data only show a major peak in the blue region between 400 and 500 nanometers.

     

Bacterial Oxidation of Sulfide Minerals in Column Leaching Experiments at Suboptimal Temperatures

The purpose of the work was to quantitatively characterize temperature effects on the bacterial leaching of sulfide ore material containing several sulfide minerals. The leaching was tested at eight different temperatures in the range of 4 to 37°C. The experimental technique was based on column leaching of a coarsely ground (particle diameter, 0.59 to 5 mm) ore sample. The experimental data were used for kinetic analysis of chalcopyrite, sphalerite, and pyrrhotite oxidation. Chalcopyrite yielded the highest (73 kJ/mol) and pyrrhotite yielded the lowest (25 kJ/mol) activation energies. Especially with pyrrhotite, diffusion contributed to rate limitation. Arrhenius plots were also linear for the reciprocals of lag periods and for increases of redox potentials (dmV/dt). Mass balance analysis based on total S in leach residue was in agreement with the highest rate of leaching at 37 and 28°C. The presence of elemental S in leach residues was attributed to pyrrhotite oxidation.     

Inhibition of Nodule Development in Soybean by Nitrate or Reduced Nitrogen

Imsande, J. 1986. Inhibition of nodule development in soybeanby nitrate or reduced nitrogen.—J. exp. Bot. 37: 348–355. Nodulation of hydroponically grown soybean plants [Glycine max(L.) Merr.] is inhibited by continuous growth in the presenceof 4· mol m^–3 KNO₃ The presence of 4·0 molm^–3 ‘starter nitrate’ for 3-6 d during noduledevelopment, however, subsequently stimulates nodule dry weightaccumulation and nitrogenase activity. These stimulations occureven though 4· mol m^–3 nitrate temporarily delaysnodule development, i.e. the late steps of nodule developmentare reversibly inhibited by a short-term exposure to 4·0mol m^–3 nitrate. On the other hand, treatment with 4·0mol m^–3 nitrate in excess of 14 d significantly reducesnodule dry weight Thus, extended growth in the presence of 4·0mol m^–3 KNO₃ seems to block both early and late stepsof nodule development. Nodulation of hydroponically grown soybeansis also inhibited by continuous growth in the presence of 2·0mol m^–3 (NH₄)₂SO₄ This inhibition is not caused by acidityof the growth medium. On the other hand, nodule development6 d after inoculation with Rhizoblum japonicum is not delayedby a 7-d exposure to 2·0 mol m^–3 (NH₄)₂SO₄ butis partially inhibited by a prolonged exposure to (NH₄)₂SO₄Because repression of nodulation by 4·0 mol m^–3KNO₃ is more severe than that by 2·0 mol m^–3 (NH₄)₂SO₄and because ammonium taken up by the soybean plant is not activelyoxidized to nitrate, it is suggested that there are at leasttwo mechanisms by which nitrate utilization represses noduleformation in soybean. Key words: Glycine max, nitrogen, nitrogen fixation, nodulation     

Nitrate Nutrition and Temperature Effects on Wheat: a Synthesis of Plant Growth and Nitrogen Uptake in Relation to Metabolic and Physiological Processes

Lawlor, D. W., Boyle, F. A., Keys, A. J., Kendall, A. C. andYoung, A. T. 1988. Nitrate nutrition and temperature effectson wheat: a synthesis of plant growth and nitrogen uptake inrelation to metabolic and physiological processes.—J.exp. Bot. 39: 329-343. Growth of spring wheat was measured in cool (13°C day/10°Cnight) or warm (23°C/18°C) temperatures, combined withlarge and small amounts of nitrate fertilizer. The rate of growthof dry matter was less at cool temperatures but total growthover the same period of development was slightly greater inthe cool than in the warm. Main-shoot and tiller leaves grewslower and, despite growing for a longer period, were shorterin the cool than in the warm. They had greater fresh and drymass and content of starch and fructosans per unit area. Coolconditions increased root dry mass, root to shoot ratio andnitrogen content in dry matter. Additional nitrate increasedleaf area of main shoots slightly but of tillers greatly; itincreased leaf and tiller dry matter and total plant dry mass.Additional nitrate decreased the proportion of dry matter inroots and in stems and the N content of dry matter in all plantparts. Regulation of growth by temperature, nitrate supply andthe rôle of photosynthesis and nitrogen uptake, is consideredin relation to the mechanisms of incorporation of carbon andnitrogen into biochemical constituents. It is concluded thattemperature regulates the rate of protein synthesis, which determinesplant growth rate. Nitrogen flux into the plant is not directlylinked to protein synthesis so that the content of NO^–₃and of amino acids is related both to growth and to conditionsgoverning NO^–₃ uptake and its reduction. When nitrogensupply is large, growth is limited by temperature, not NO^–₃.Inadequate nitrate supply decreases protein synthesis (and thereforegrowth) more than it decreases carbon assimilation, so thatorgans such as roots and stems increase in dry matter relativeto shoots and all tissues have smaller proportions of nitrogenin dry matter. Cool conditions, although decreasing the rateof protein synthesis, increase its duration and decrease thesize of leaves, so that the content of protein per unit leafarea is greater in cool than in warm grown leaves. Consequencesof changes in the balance of N and C supply and growth ratefor dry matter distribution in plants are discussed. Key words: Wheat, nitrate nutrition, temperature     

Cyclic Variations in Nitrogen Uptake Rate of Soybean Plants: Effects of External Nitrate Concentration

Net uptake of by non-nodulated soybean plants [Glycine max (L.) Merr.cv. Ransom] growing in flowing hydroponiccultures containing 0–5, 1.0 and 10-0 mol m^–3 was measured daily during a 24-d period of vegetativedevelopment to determine if amplitude of maximum and minimumrates of net uptake are responsive to external concentrations of . Removal of from the replenished solutions during each 24-h period was determinedby ion chromatography. Neither dry matter accumulation nor theperiodicity of oscillations in net uptakerate was altered bythe external concentrations. The maxima of the oscillations in net uptake rate, however, increased nearly3-fold in response to external concentrations. The maxima and minima, respectively, changed from 4.0 and 0–6mmol per gram root dry weight per day at an external solution level of 05 mol m^–3 to 15–2and -2–7 mmol per gram root dry weight per day at an external solution level of 10–0mol m^–3 . The negative values for minimum net uptake rate from10–0 mol m^–3 solutions show that net efflux was occurring andindicate thatthe magnitude of the efflux component of net uptake was responsiveto external concentration of . Key words: Nitrate uptake rate, carbon-nitrogen partitioning, flowing solution culture     

Relationship between Nitrate Uptake, Flux, and Reduction and the Accumulation of Reduced Nitrogen in Maize (Zea mays L.): I. GENOTYPIC VARIATION

The study presented here was an extension of a preceding field project concerned with changes in N metabolism of four maize hybrids during grain development. The objectives were to relate uptake, flux, and reduction of nitrate to accumulation of reduced N in growth-chamber-grown seedlings of the same four hybrids and to compare these results with those obtained in the field study.     

Rates of Digestion of Bacteria by Marine Phagotrophic Protozoa: Temperature Dependence

The effect of temperature on length of time for digestion of bacteria was evaluated, by using fluorescently labeled bacteria (FLB), for phagotrophic flagellates and ciliates isolated from coastal northwest Mediterranean waters. Accumulation of FLB in protozoan food vacuoles was followed until a plateau of FLB per cell occurred; then after a 1:10 dilution of FLB with unlabeled bacteria, disappearance of FLB in food vacuoles was monitored. For both 3- to 5-μm flagellates and 10- to 40-μm ciliates, the absolute linear slopes of FLB uptake and disappearance were nearly identical in individual experiments over a temperature range of 12 to 22°C. We inferred from these results that the leveling off of the uptake curves resulted when equilibrium between ingestion and digestion of bacteria was attained. The time to leveling off then represented the average time needed for complete digestion of the bacteria ingested at the start of the experiment, and the inverse of this time represented a bacterial digestion rate. The digestion rate increased exponentially from 12 to 22°C for both a mixed flagellate assemblage and the oligotrichous ciliate Strombidium sulcatum, with a Q₁₀ of 2.8 for the flagellates and 2.0 for the ciliate. Although bacterial ingestion rates varied greatly, depending on protozoan cell size, total bacterial abundance, and temperature, digestion times appeared to be significantly influenced only by protozoan cell size (or type of protozoan) and by temperature.     

Nitrogen Utilization in Lemna: II. Studies of Nitrate Uptake Using NO(3)

¹³N-labeled nitrate was used to trace short-term nitrate influx into Lemna gibba L. G3 in experiments where disappearance of both radioactivity and total nitrate from the incubation medium was measured continuously and simultaneously. In plants performing net nitrate uptake from an initial nitrate concentration of 40 to 60 micromolar, there was no discrepancy between net uptake and influx, irrespective of the N status of the plants, indicating that concomitant nitrate efflux was low or nil. Plants treated with tungstate to inactivate nitrate reductase were able to take up nitrate following induction of the uptake system by exposure to a low amount of nitrate. Also, in this case, net uptake was equivalent to influx. In tungstate-treated plants preloaded with nitrate, both net uptake and influx were nil. In contrast to these observations, a clear discrepancy between net uptake and influx was observed when the plants were incubated at an initial nitrate concentration of approximately 5 micromolar, where net uptake is low and eventually ceases. It is concluded that plasmalemma nitrate transport is essentially unidirectional in plants performing net uptake at a concentration of 40 to 60 micromolar, and that transport is nil when internal nitrate sinks (vacuole, metabolism) are eliminated. The efflux component becomes increasingly important when the external concentration approaches the threshold value for net nitrate uptake (the nitrate compensation point) where considerable exchange between internal and external nitrate occurs.     

Concurrent Rates of N2 Fixation, Nitrate and Ammonium Uptake by White Clover in Response to Growth at Different Root Temperatures

Nodulated white clover plants (Trifolium repens L. cv. Huia)were grown for 71 d in flowing nutrient solutions containingN as 10 mmol m_–3 NH₄NO₃, under artificial illumination,with shoots at 20/15°C day/night temperatures and root temperaturereduced decrementally from 20 to 5°C. Root temperatureswere then changed to 3, 7, 9, 11, 13, 17 or 25°C, and theacquisition of N by N₂ fixation, NH₄+ and NO₃^– uptakewas measured over 14 d. Shoot specific growth rates (d. wt)doubled with increasing temperature between 7 and 17°C,whilst root specific growth rates showed little response; shoot:root ratios increased with root temperature, and over time at11°C. Net uptake of total N per plant (N₂ fixation + NH₄++ NO₃^–) over 14 d increased three-fold between 3 and 17°C.The proportion contributed by N₂ fixation decreased with increasingtemperature from 51% at 5°C to 18% at 25°C. Uptake ofNH₄+ as a proportion of NH₄+ + NO₃^– uptake over 14 d variedlittle (55–62%) with root temperature between 3 and 25°C,although it increased with time at most temperatures. Mean ratesof total N uptake per unit shoot f. wt over 14 d changed littlebetween 9 and 25°C, but decreased progressively with temperaturebelow 9°C, due to the decline in the rates of NH₄+ and NO₃^–uptake, even though N₂ fixation increased. The results suggestthat N₂ fixation in the presence of sustained low concentrationsof NH₄+ and NO₄^– is less sensitive to low root temperaturethan are either NH₄+ or NO₃^– uptake systems. White clover, Trifolium repens L. cv. Huia, root temperature, nitrogen fixation, ammonium, nitrate     

Plant Growth Promoting Rhizobacteria and Soybean [ Glycine max (L.) Merr.] Nodulation and Nitrogen Fixation at Suboptimal Root Zone Temperatures

Co-inoculation of plant growth promoting rhizobacteria (PGPR)withBradyrhizobium has been shown to increase legume nodulationand nitrogen fixation at optimal soil temperatures. Nine rhizobacteriaco-inoculated withBradyrhizobium japonicum532C were tested fortheir ability to reduce the negative effects of low root zonetemperature (RZT) on soybean [Glycine max(L.) Merr.] nodulationand nitrogen fixation. Three RZTs were tested: 25 (optimal),17.5 (somewhat inhibitory), and 15°C (very inhibitory).At each temperature some PGPR strains increased the number ofnodules formed and the amount of fixed nitrogen when co-inoculatedwithB. japonicum,but the stimulatory strains varied with temperatures.The strains that were most stimulatory varied among temperaturesand were as follows: 15°C,Serratia proteamaculans 1-102;17.5°C,S. proteamaculans 1-102andAeromonas hydrophilaP73;25°C,Serratia liquefaciens2-68. Bradyrhizobium japonicum ; Glycine max; plant growth promoting rhizobacteria; suboptimal root zone temperatures     

Growth of Bacteria Degrading Naphthalene and Salicylate at Low Temperatures

A total of 58 bacterial strains degrading naphthalene and salicylate were isolated from soil samples polluted with oil products, collected in different regions of Russia during winter and summer. The isolates were assessed for their ability to grow at low temperatures (4, 8, and 15°C); bacteria growing at 4°C in the presence of naphthalene or salicylate accounted for 65 and 53%, respectively, of the strains isolated. The strains differed in the temperature dependence of their growth rates. It was demonstrated that the type of expression of the Nah⁺ phenotype at low temperatures depended on the combination of host bacterium and plasmid.     

Isolation of Cave Bacteria and Substrate Utilization at Different Temperatures

Bacteria isolated from the soils of three northern Spain caves were investigated for their growth at temperatures in the range 5-45°C, substrate utilization pattern, and cellular fatty acid components. All tested bacteria were able to grow comparatively well in a range of temperatures from 13° to 45°C. At 13°C the bacteria oxidized on average 87% of carbon sources provided while incubation at 28°C reduced to about 50% carbon source utilization. Changes in fatty acid profiles reflected an adaptative response to temperature. The results suggest the need of using low temperatures to detect maximum diversity of culturable bacteria (other than actinomycetes) and efficiency of carbon utilization by cave bacteria.     

A Temperature Dependence of the Intra- and Extracellular Fatty-Acid Composition of Green Algae and Cyanobacterium

The effect of ambient temperature on the composition of intracellular fatty acids and the release of free fatty acids (FFA) into a medium by cyanobacterium Spirulina platensis and eukaryotic microalgae, Chlorella vulgaris and Botryococcus braunii, was studied using their batch cultures. It was found that all the species studied, regardless of their taxonomic status, responded to the temperature regime by similar changes in their intracellular fatty acid composition: the relative content of more unsaturated fatty acids decreased with the elevation of temperature. At the same time, in the prokaryote, this temperature shift blocked, first of all, the elongation of 16:0 to 18:0 and then their further desaturation. In eukaryotes, the change in the desaturation of dienoic to trienoic fatty acids was the most pronounced process. The ratio of dienoic to trienoic fatty acids remained almost unchanged in S. platensis. The relative content of extracellular unsaturated FFA increased in the prokaryotic organism S. platensis at a higher temperature. But no significant changes in the composition of extracellular unsaturated FFA were detected in eukaryotic algae upon temperature elevation.