Productivity of outdoor algal cultures in unstable weather conditions

In the outdoor cyclic fed batch cultures of Chlorella pyrenoidosa, some typical growth kinetics patterns in unstable weather conditions were observed. On cloudy days, the biomass output rate (R) was low, but the bioenergetic growth yield (Y) was generally high. In the cloudy morning-sunny afternoon condition, the values of Y were low, especially in the afternoon. In the sunny morning-cloudy afternoon condition, both R and Y were high. A few very high short-term Y values were measured during the cloudy the cloudy afternoon. (c) 1993 Wiley & Sons, Inc.     

On the applicability of adaptive bioprocess state estimators

This article presents an industrial case study, examining the application of a novel adaptive biomass estimator to an industrial microfungi production process. It is our intention that this contribution should focus upon the implementation issues of the algorithm, in preference to a rigorous theoretical development. The novel algorithm adopted is developed from Adaptive Inferential Estimation studies of Guilandoust and co-workers. The technique utilizes input-output process measurements obtained at different frequencies, thereby providing more frequent estimates of biomass concentration than are otherwise available from off-line laboratory analyses. The algorithm is particularly suited to the biotechnology industry, as it is capable of utilizing irregular assay measurements with varying delays.Although this article demonstrates the encouraging industrial implications of the adaptive algorithm, like all adaptive techniques currently developed, it is restricted by the inability to perform robust on-line system identification. The ultimate selection of a "suboptimal" "fixed parameter" algorithm for on-line implementation, is therefore directly attributable to these inadequacies. Aspects of data acquisition, data pretreatment, and data quality are critical for real process applications, and while some practical approaches are adopted here, many important implementation problems remain unresolved. (c) 1993 John Wiley & Sons, Inc.     

Use of ATP to characterize biomass viability in freely suspended and immobilized cell bioreactors

This work describes investigations into the viability of cells growing on 3,4-dichloroaniline (34DCA). Two bioreactors are employed for microbial growth, a continuous stirred tank (CST) bioreactor with a 2-L working volume, and a three-phase air lift (TPAL) bioreactor with a 3-L working volume. Experiments have been performed at several dilution rates between 0.027 and 0.115 h(-1) in the CST bioreactor and between 0.111 and 0.500 h(-1) in the TPAL bioreactor. The specific ATP concentration was calculated at each dilution rate in the suspended biomass in both bioreactors as well as in the immobilized biomass in the TPAL bioreactor. The ATP was extracted from the cells using boiling tris-EDTA buffer (pH 7.75), and the quantity determined using a firefly (bioluminescence) technique. The cultures were inspected under an electron microscope to monitor compositional changes. Results from the CST bioreactor showed that the biomass-specific ATP concentration increases from 0.44 to 1.86 mg ATP g(-1) dry weight (dw) as dilution rate increases from 0.027 to 0.115 h(-1). At this upper dilution rate the cells were washed out. The specific ATP concentration reached a limiting average value of 1.73 mg ATP g(-1) dw, which is assumed to be the quantity of ATP in 100% viable biomass. In the TPAL bioreactor, the ATP level increased with dilution rate in both the immobilized and suspended biomass. The specific ATP concentration in the immobilized biomass increased from approximately 0.051 mg ATP g(-1) dw at dilution rates between 0.111 and 0.200 h(-1) to approximately 0.119 mg ATP g(-1) dw at dilution rates between 0.300 and 0.500 h(-1). This indicates that the immobilized biomass contained a viable cell fraction of around 5%. Based on these results, kinetic data for freely suspended cells should not be applied to the modeling of immobilized cell systems on the assumption that immobilized biomass is 100% viable. (c) 1993 John Wiley & Sons, Inc.     

Seasonal variation in the physical properties of agar and biomass of Gracilaria sp. (chorda type) from Tosa Bay,southern Japan

Plants of Gracilaria sp.(chorda type), which grow along the coast of Uranouchi Inlet in Tosa Bay, southern Japan, showed the highest biomass in the summer (26 °C to 31 °C) and spring season (15.1 °C to 24.9 °C). Maximum biomass was 6952 g m^–2 in July, but gradually decreased in the autumn (30.5 °C in September to 20 °C in November) and winter (19.5 °C in December to 14.9 °C in February). Variation in yields and gel strength of the agars, were shown to depend on the time in the season. After alkali treatment (5% NaOH, 2 h) at three different temperatures (70, 80, and 90 °C), the agars showed gel strengths essentially that of commercial grade agars, with the best gel obtained at 80 °C. Maximum gel strength (1455 g cm^–2 of 1.5% agar gel) occurred in winter when the biomass and agar yield were low. Minimum gel strength was in spring. Gel strength was inversely correlated with agar yield, but was positively correlated with apparent viscosity. Maximum viscosity was 40 cP. in December. Gelling temperatures, pH of 1.5% agar gel, and moisture content in agars showed little variation.     

Seasonal succession of phytoplankton in a lake of the Paraná river floodplain,Argentina

Phytoplankton biomass, morphological and taxonomic composition, species diversity and productivity were analyzed in a shallow lake of the Middle Paraná River floodplain (El Tigre, 31 ° 41 S and 60° 42 W), between November 1986 and July 1988. Lake inundation (filling and through-flow phases) constituted an intense long-term perturbation in the physical and chemical environment. As the lake filled with river water, K-selected species (netplanktonic filamentous bluegreens, > 37 µm, with low surface area/volume (SA/V) ratios) that had existed prior to filling (late spring 1986) were replaced in summer-fall by r-selected species (nannoplanktonic chlorophytes and cryptophytes, < 37 µm, mainly stout forms with high SA/V ratios). During the through-flow phase, lentic phytoplankton was replaced by lotic flagellate populations due to the direct flushing by river water. During the period of falling water (drainage and isolation phases), nanoplanktonic algae with similar characteristics to those of the filling phase dominated in late winter-spring. Later in the isolation phase, these were succeeded by K-selected species (netplanktonic algae, mainly motile spherical dinoflagellates and filamentous bluegreens with low SA/V ratios). Simultaneously, primary production per unit biomass decreased and total biomass and specific diversity increased. Seasonal changes of phytoplankton in floodplain lakes can be interpreted as the interaction between true successional development (as observed in the drainage and isolation phases) and intermediate disturbance. Using Reynolds' terminology, short-term disturbance (slight inflow of nutrient-rich river water) caused reversion to an earlier stage in the former succession, and long-term disturbance (lake inundation) truncated the successional progression and a new (or shifted) succession was initiated.     

Location of the major barriers to water and ion movement in young roots of Zea mays L.

The main barriers to the movement of water and ions in young roots of Zea mays were located by observing the effects of wounding various cell layers of the cortex on the roots' hydraulic conductivities and root pressures. These parameters were measured with a root pressure probe. Injury to the epidermis and cortex caused no significant change in hydraulic conductivity and either no change or a slight decline in root pressure. Injury to a small area of the endodermis did not change the hydraulic conductivity but caused an immediate and substantial drop in root pressure. When large areas of epidermis and cortex were removed (15–38% of total root mass), the endodermis was always injured and root pressure fell. The hydraulic conductance of the root increased but only by a factor of 1.2–2.7. The results indicate that the endodermis is the main barrier to the radial movement of ions but not water. The major barrier to water is the membranes and apoplast of all the living tissue. These conclusions were drawn from experiments in which hydrostatic-pressure differences were used to induce water flows across young maize roots which had an immature exodermis and an endodermis with Casparian bands but no suberin lamellae or secondary walls. The different reactions of water and ions to the endodermis can be explained by the huge difference in the permeability of membranes to these substances. A hydrophobic wall barrier such as the Casparian band should have little effect on the movement of water, which permeates membranes and, perhaps, also the Casparian bands easily. However, hydrophobic wall depositions largely prevent the movement of ions. Several hours after wounding the endodermis, root pressure recovered to some extent in most of the experiments, indicating that the wound in the endodermis had been partially healed.Abbreviations Lp_r hydraulic conductivity of root; T_1/2 = half-time of water exchange between root xylem and external medium This research was supported by a grant from EUROSILVA (project no. 39473C) to E.S., and by a Bilateral Exchange Grant jointly funded by the Deutsche Forschungsgemeinschaft and the Natural Sciences and Engineering Research Council of Canada to C.A.P. We thank Mr. Burkhard Stumpf for his excellent technicial assistance.     

Glycolytic enzymes in non-photosynthetic plastids of pea (Pisum sativum L.) roots

The presence of the glycolytic enzymes from hexokinase to pyruvate kinase in plastids of seedling pea (Pisum sativum L.) roots was investigated. The recoveries, latencies and specific activities of each enzyme in different fractions was compared with those of organelle marker enzymes. Tryptic-digestion experiments were performed on each enzyme to determine whether activities were bound within membranes. The results indicate that hexokinase (EC 2.7.1.2) and phosphoglyceromutase (EC 5.4.2.1) are absent from pea root plastids. The possible function of the remaining enzymes is considered.Abbreviations GADPH glyceraldehyde 3-phosphate dehydrogenase - PFK phosphofructokinase - PFP pyrophosphate: fructose 6-phosphate 1-phosphotransferase Bronwen A. Trimming gratefully acknowledges the award of a studentship from the Science and Engineering Research Council     

Effect of temperature and photoperiod on the raffinose content of spruce roots

The influence of temperature and photoperiod on raffinose synthesis in spruce roots (Picea abies (L.) Karst.) was investigated under controlled environmental conditions in a phytotron. The raffinose content of the roots increased when the plants were subjected simultaneously to a change from long to short days and from summer-like day and night temperatures to a climate which was more than 10° C colder. Only a very slight raffinose accumulation resulted from a change of day-length or temperature alone, but a subsequent additional change of temperature or daylength, respectively, caused an increase in the raffinose content, yet only to half the amount found when both climate factors changed simultaneously. When the shoot was left under non-inducing conditions, but the root was cooled, the raffinose content increased in the root, but not in the shoot. The root was also capable of inducing raffinose if the shoot was cut off after a few days of cold adaptation of the whole plant. For all climate changes the sucrose content changed much less than the raffinose content. Induction of raffinose was comparable in mycorrhizal and in non-mycorrhizal roots.Abbreviations DW dry weight - LDC long day, cold - LDW long day, warm - SDC short day, cold - SDCLL short day, cold, low light - SDF short day, frost - SDW short day, warm This research was supported by the Bundesamt für Bildung und Wissenschaft and by the Swiss National Science Foundation.     

Studies on the behavior of organelles and their nucleoids in the root apical meristem of Arabidopsis thaliana (L.) Col.

The behavior of cell nuclei, mitochondrial nucleoids (mt-nucleoids) and plastid nucleoids (ptnucleoids) was studied in the root apical meristem of Arabidopsis thaliana. Samples were embedded in Technovit 7100 resin, cut into thin sections and stained with 4′-6-diamidino-2-phenylindole for light-microscopic autoradiography and microphotometry. Synthesis of cell nuclear DNA and cell division were both active in the root apical meristem between 0 μm and 300 μm from the central cells. It is estimated that the cells generated in the lower part of the root apical meristem enter the elongation zone after at least four divisions. Throughout the entire meristematic zone, individual cells had mitochondria which contained 1–5 mt-nucleoids. The number of mitochondria increased gradually from 65 to 200 in the meristem of the central cylinder. Therefore, throughout the meristem, individual mitochondria divided either once or twice per mitotic cycle. By contrast, based on the incorporation of [³H]thymidine into organelle nucleoids, syntheses of mitochondrial DNA (mtDNA) and plastid DNA (ptDNA) occurred independently of the mitotic cycle and mainly in a restricted region (i.e., the lower part of the root apical meristem). Fluorimetry, using a videointensified microscope photon-counting system, revealed that the amount of mtDNA per mt-nucleoid in the cells in the lower part of the meristem, where mtDNA synthesis was active, corresponded to more than 1 Mbp. By contrast, in the meristematic cells just below the elongation zone of the root tip, the amount of mtDNA per mt-nucleoid fell to approximately 170 kbp. These findings strongly indicate that the amount of mtDNA per mitochondrion, which has been synthesized in the lower part of the meristem, is gradually reduced as a result of continual mitochondrial divisions during low levels of mtDNA synthesis. This phenomenon would explain why differentiated cells in the elongation zone have mitochondria that contain only extremely small amounts of mtDNA. This work was supported by a Grant-in Aid (T.K.) for Special Research on Priority Areas (Project No. 02242102, Cellular and Molecular Basis for Reproduction Processes in Plants) from the Ministry of Education, Science and Culture of Japan and by a Grant-in Aid (T.K.) for Original and Creative Research Project on Biotechnology from the Research Council, Ministry of Agriculture, Forestry and Fisheries of Japan.     

Al3+-Ca2+ interactions in aluminum rhizotoxicity

Several mineral rhizotoxicities, including those induced by Al³⁺, H⁺, and Na⁺, can be relieved by elevated Ca²⁺ in the rooting medium. This leads to the hypothesis that the toxic cations displace Ca²⁺ from transport channels or surface ligands that must be occupied by Ca²⁺ in order for root elongation to occur. In this study with wheat (Triticum aestivum L.) seedlings, we have determined, in the case of Al³⁺, that (i) Ca²⁺, Mg²⁺, and Sr²⁺ are equally ameliorative, (ii) that root elongation does not increase as Ca²⁺ replaces Mg²⁺ or Sr²⁺ in the rooting media, and (iii) that rhizotoxicity is a function solely of Al³⁺ activity at the root-cell membrane surface as computed by a Gouy-Chapman-Stern model. The rhizotoxicity was indifferent to the computed membrane-surface Ca²⁺ activity. The rhizotoxicity induced by high levels of tris(ethylenediamine)cobaltic ion (TEC³⁺), in contrast to Al³⁺, was specifically relieved by Ca²⁺ at the membrane surface. The rhizotoxicity induced by H⁺ exhibited a weak specific response to Ca²⁺ at the membrane surface. We conclude that the Ca²⁺-displacement hypothesis fails in the case of Al³⁺ rhizotoxicity and that amelioration by cations (including monovalent cations) occurs because of decreased membrane-surface negativity and the consequent decrease in the membrane-surface activity of Al³⁺. However, TEC³⁺, but not Al³⁺, may be toxic because it inhibits Ca²⁺ uptake. The nature of the specific H⁺-Ca²⁺ interaction is uncertain.Abbreviations {Al³⁺ }₀ chemical activity of Al³⁺ at the root-cell membrane surface - {Al³⁺ }_E chemical activity of Al³⁺ in the external rooting medium - E₀ electrical potential at the root-cell membrane surface - HXM²⁺ hexamethonium ion - TEC³⁺ tris(ethylenediamine)cobaltic ion