Separation and Purification of Molecular Species of Galactolipids by High Performance Liquid Chromatography

The NaCl concentration of the growth medium affected hydrogen production by Lyngbya sp. (No. 108) strain. Cells grown in medium containing 3% NaCl produced the most hydrogen. The carbohydrate content of this strain also increased with increasing NaCl concentration of the growth medium up to 720 fig/mg cells at 5 % NaCl. In the presence of 20 finlol/ml MFA (monofluoroacetic acid), inhibition of hydrogen production was observed. We extracted the glycogen from this nonheterocystous filamentous cyanobacterium, Lyngbya sp. (No. 108), and observed that glycogen and carbohydrate consumption of this strain is coincident with hydrogen production.

These results led us to the conclusion that the reserve glycogen or other carbohydrate were used as sources of electron donors for hydrogen production, and that the NaCl concentration of the medium affected the hydrogen production by this strain.     

Phosphate Metabolism in the Cyanobacterium Anabaena doliolum Under Salt Stress

In the present study, we have investigated the effects of NaCl concentrations on the growth and phosphate metabolism of an Anabaena doliolum strain isolated from a paddy field, in order to determine the possible effects of salinization. Growth rate, chlorophyll content, and protein content decreased with increasing salt concentration in the growth medium, while carbohydrate concentration increased. Phosphate content and phosphate uptake rate decreased. There was an increase in total alkaline phosphatase activity, with an approximately 7-fold increase in extracellular activity compensating for an approximately 3-fold decrease in cell-bound activity. NaCl effects on protein and chlorophyll concentrations were greater in P-deficient medium, while presence or absence of P in the medium had little effect on cellular carbohydrate concentrations. It is concluded that growth in high salt likely leads to reduced phosphate uptake in A. doliolum.     

Polyethylene glycol and maltose enhance somatic embryo maturation in loblolly pine (Pinus taeda L.)

Summary A culture medium that can efficiently produce mature somatic embryos was developed for loblolly pine (Pinus taeda L.). The medium contained maltose as a carbohydrate source and polyethylene glycol as an osmoticum. This medium formulation significantly enhanced embryo maturation efficiency compared to a medium with only maltose, or with sucrose combined with polyethylene glycol. Maltose at 4% and polyethylene glycol at 6% resulted in the highest embryo maturation efficiency; an average of around 100 cotyledonary embryos were produced from 1 g of embryogenic tissue. These results suggested that previous ineffective embryo maturation in loblolly pine may be due to the lack of the proper combination of osmoticum and carbohydrate source. This embryo maturation method also improved morphology of cotyledonary embryos of loblolly pine.     

The influence of medium on the chemical composition ofBlastomyces dermatitidis

Whole cells and cell walls of the mycelial and yeast forms ofBlastomyces dermatitidis grown in four different media were analyzed for differences in lipid, fatty acid, carbohydrate, and protein contents. The bound (saponifiable) fatty acids of yeast and mycelial whole cells (but not the cell walls) vary considerably in response to growth medium. The percentage of readily extractable lipid varied somewhat in whole cells. The percentage of carbohydrate and protein of whole cells and cell walls are little affected by the medium in which the cells are grown.     

Milbemycin production byStreptomyces sp.: the effect of carbohydrates

Summary Milbemycin production byStreptomyces hygroscopicus RB4569D was examined in media containing different carbohydrates. Total milbemycin titer could be increased by substitution of fructose for glucose and by selection of the appropriate starch type. Total titer could be further enhanced by increasing the concentration of fructose and/or starch in the medium. Rates of carbohydrate utilization were shown to be independent of their initial concentration and increased titers in high carbohydrate media were shown to be due to a prolonged production phase rather than an increased accretion rate. The pattern of individual milbemycin components was governed by the carbon:nitrogen ratio of the medium rather than carbohydrate concentration and there was a critical C:N ratio below which no milbemycin was produced.     

Carbohydrate Uptake and Metabolism of Ophiobolus graminis

The carbohydrate nutrition of Ophiobolus graminis, the cause of the take-all disease of wheat, was investigated in growth and respiration experiments. In a synthetic medium, d-mannitol was the only carbohydrate of thirteen studied which the fungus could not use for growth. However, the fungus was found to take up mannitol by an active mechanism, which was stopped by 2,4-dinitrophenol. Di- and trisaccharides were hydrolyzed extracellularly, and the monosaccharides were assimilated at different rates.     

Apparent extracellular glycoprotein turnover product from Candida albicans

Low molecular weight material was isolated from the culture medium of three strains of Candida albicans. This material was produced from exponentially growing cultures and it appeared from chemical analysis to represent the carbohydrate portion of cell surface glycoproteins. The material contained a few residual amino acids which were interpreted to represent the attachment points of carbohydrate to protein and the remanents protease cleavage sites. Some of this material could be generated artificially by treating intact cells with papain.     

The Physiology of Rooting Populus Cuttings

The changing levels of carbohydrates and the rates of photosynthesis in rooting Populus cuttings are described. The cuttings were planted in a sand medium, in controlled environment conditions. There was no evidence to support the view that failure to root in P. tremula was caused by insufficient carbohydrate reserves. In leafless hardwood cuttings, the carbohydrate levels were initially very high (14–19%) but fell rapidly (to 5–10%) as roots, callus or shoots developed. In leafy softwood cuttings, a steady accumulation of carbohydrate reserves was observed, rising from 5–10% on planting to 15–25% when roots had developed. One of the differences between the easy-to-root P. × euramericana and the hard-to-root P. tremula was the apparent downward transport of assimilates in P. × euramericana softwood cuttings.     

Factors Affecting Somatic Embryogenesis Induction and Conversion in “Paradise Tree” (<Emphasis Type="Italic">Melia azedarach</Emphasis> L.)

Factors affecting somatic embryogenesis induction and conversion in paradise tree (Melia azedarach) were evaluated. Somatic embryogenesis was influenced by plant growth regulators, explant stage, carbohydrate source and concentration, gelling agents, light, and induction times. MS medium with 4.54 μM thidiazuron (TDZ) was optimal for the induction of embryogenic tissue. Zygotic embryos that were 1-1.5 mm long (torpedo and early cotyledonal stage) had a greater embryogenic response than smaller or larger embryos and better conversion of somatic embryos into plants. In general, embryos that formed in medium containing 1% or 5% carbohydrate were hyperhydrics or fused, respectively, whereas those that formed in medium with a carbohydrate concentration of 3% had better morphology. Raffinose at 3% yielded satisfactory somatic embryo induction with good morphology and the best values of conversion into plants. Induction and conversion of somatic embryos were superior on medium solidified with agar A-1296. The explants maintained under 160 μmol m⁻² s⁻¹ or 1 week in darkness and later 160 μmol m⁻² s⁻¹ produced a significantly higher embryogenic index. Only 4 days of treatment on induction medium, with either raffinose or sucrose at 3% as a carbohydrate source, were required to induce somatic embryogenesis, but longer exposure, until 18 days, increased the yield and improved the morphology of somatic embryos.     

Studies on the Metabolism of Aspergillii

Endogenous respiration of Aspergillus sojae K.S. was studied in terms of biochemical analysis. It was found that the different kind of substrates was utilized for the endogenous respiratoin according to C:N ratio of the agar medium on which the mold was grown. In the mycelial mats grown on the medium of low C:N value, pool amino acids, protein, and nucleic acids were mainly utilized from the beginning while carbohydrate or lipid displayed a minor role. The corresponding amount of ammonia was formed. On the other hand, in the mycelial mats grown on the medium of rather high C:N value, carbohydrate or lipid was the major substrate of endogenous respiration in the early stages of incubation. The utilization of the nitrogenous materials and the accompanying formation of ammonia got to start only after the lapse of several hours of incubation.     

Metabolic Changes in Trichomonas gallinae Resulting from Growth in Various Carbohydrates*†

SYNOPSIS. The influence of the type of growth carbohydrate on the subsequent metabolic activity of Trichomonas gallinae was investigated. Washed suspensions of cells collected from CPL-glucose, CPL-maltose, CPL-galactose, and CPL-glucose-maltose media were examined in the Warburg respirometer for their ability to utilize glucose, maltose, and galactose. Comparisons of the metabolic parameters of substrate consumption, changes in glycogen content, and CO₂ and H₂ production were made. The pattern of utilization of the sugars, both qualitatively and quantitatively, depended upon the type of carbohydrate in the CPL medium used to culture the cells and upon the time of exposure of the cells to a particular sugar in the medium.     

Wachstum und N-Stoffwechsel sowie Anderungen des Isoenzymspektrums der Glutamatdehydrogenase (GDH) in batchvermehrten, licht- und dunkelkultivierten Zellsuspensionen von Pisum sativum

Growth, N-metabolism and isoenzyme pattern of glutamate dehydrogenase in batch-cultures of Pisum sativum cells under light and dark conditions. Cell suspension cultures of Pisum sativum L. derived from root and shoot sections of seedlings have been prepared and cultured in defined nutrient medium. Both the cells and the media were analysed daily for the N-fractions and carbohydrates during the growth period. The data obtained indicate specific correlations between growth and nitrogen and carbohydrate metabolism. At the beginning of the growth cycle ammonia as compared to nitrate was favoured in uptake. An increased uptake of nitrate occurred at the end of the linear growth phase when carbohydrate in the media was depleted. The uptake of sucrose was rapid during the whole growth cycle, only in the range of the linear growth phase the uptake stagnated for 3 or 4 days. During increased biosynthesis of nitrogenous compounds at the beginning of the growth cycle up to seven isoenzymes of the glutamate dehydrogenase could be separated by polyacrylamide gel electrophoresis. The isoenzyme pattern changed during the stationary growth phase, especially when the carbohydrate content in the medium decreased. There is some evidence that the isoenzyme pattern is influenced by carbohydrate metabolism.     

Degradation of food compounds and growth response on different food quality by the anaerobic ciliate Trimyema compressum

The biochemical composition of two food bacteria was examined on which monoxenic cultures of Trimyema compressum grew with different yields. The food bacteria were the saccharolytic fermenting bacterium Bacteroides WoCb15 and the purple nonsulfur bacterium Rubrivivax gelatinosus. Differences in composition of bacterial biomass concerned mainly the carbohydrate content. By different culture conditions for R. gelationsus and pasteurization of carbohydrate-rich cells, we were able to feed the ciliate with food mixtures of different carbohydrate content. Dry mass yields of the ciliate reached a maximum with mixtures of 80% carbohydrate-rich pasteurized cells plus 20% carbohydratepoor living cells. In the absence of degradable carbohydrate, energy metabolism depended on protein as substrate. Utilization of protein was incomplete, large amounts were converted into soluble compounds that accumulated in the culture medium. The ciliate consumed storage carbohydrate of living or pasteurized food bacteria equally well, while growth with short generation times was still dependent on a certain percentage of living bacteria as source of native protein. Lipids, nucleic acids and denatured proteins were not degradable by the ciliate. Consequences for the fermentative metabolism of Trimyema compressum are discussed.     

Interrelationships between carbohydrate metabolism and nitrogen assimilation in cultured plant cells

Summary The effect of the nature and concentration of the nitrogen source on respiratory activity and removal of carbohydrate from the medium in suspension cultured sycamore (Acer pseudoplatanus L.) cells was determined. Comparison was also made of the rates of uptake of the two alternative nitrogen sources, nitrate and glutamate, at differing initial nitrogen concentrations within the range 7–14 mM. The initial pH of the culture medium before inoculation was 5.2; after inoculation the pH of both nitrate and glutamate cultures rose to reach an eventual level in the range 7.0–7.1. Glutamate was removed from the medium more slowly than nitrate. Under the particular conditions of culture used the growth of the cells was nitrogen limited. Sugar uptake from the medium continued for some time after the nitrogen in the medium was depleted. The data show that although cell division and protein content are nitrogen-limited, dry weight and fresh weight yields may also be determined in a complex interaction through carbohydrate availability. There were no obvious differences in respiratory activity between cultures grown on nitrate or glutamate.     

Properties of a Novel Extracellular Cell-free Ice Nuclei from Ice-nucleating Pseudomonas antarctica IN-74

Some ice-nucleating bacterial strains, including Pantoea ananatis (Erwinia uredovora), Pseudomonas fluorescens, and Pseudomonas syringae isolates, were examined for the ability to shed ice nuclei into the growth medium. A novel ice-nucleating bacterium, Pseudomonas antarctica IN-74, was isolated from Ross Island, Antarctica. Cell-free ice nuclei from P. antarctica IN-74 were different from the conventional cell-free ice nuclei and showed a unique characterization. Cell-free ice nuclei were purified by centrifugation, filtration (0.45 μm), ultrafiltration, and gel filtration. In an ice-nucleating medium in 1 liter of cell culture, maximum growth was obtained with the production of 1.9 mg of cell-free ice nuclei. Ice nucleation activity in these cell-free ice nuclei preparations was extremely sensitive to pH. It was demonstrated that the components of cell-free ice nuclei were protein (33%), saccharide (12%), and lipid (55%), indicating that cell-free ice nuclei were lipoglycoproteins. Also, carbohydrate and lipid stains showed that cell-free ice nuclei contained both carbohydrate and lipid moieties.     

Biostimulant activity of humic-like substances from agro-industrial waste on Chlorella vulgaris and Scenedesmus quadricauda

The use of microalgae in a number of sectors, including biodiesel, feed and food production, is proving to be of great interest. An evaluation was made of the possible biostimulant effects on Chlorella vulgaris and Scenedesmus quadricauda of humic-like substances (HLs) extracted from agro-industrial wastes. These included digestate from the waste of an agro-livestock farm (D-HL), oil extraction residues from rape (B-HL, Brassica napus L.) and tomato residues (T-HL). The microalgal growth medium (BG11) was supplemented with HLs to evaluate their effect on biomass yield as well as carbohydrate, chlorophylls a and b, lipid and fatty acid contents. Our results showed that the HLs used in the test are effective biostimulants of C. vulgaris and S. quadricauda. The biostimulant effect seems to depend on the type of extract used for cultivating the microalgae, the concentration and the species treated. Among the extracts applied to the growth medium, D-HL and T-HL seem to have a significant effect on microalgal biomass and lipid production. Although B-HL showed no significant effect on the biomass and lipid content of C. vulgaris and S. quadricauda, its presence in the growth medium increased the saturated:unsaturated fatty acid ratio (SFA/UFA) and stimulated the sugar metabolism of the microalgae by increasing their carbohydrate and chlorophyll content.     

Studies on the growth and encystment ofPolytomella agilis

Summary Changes in cell population density, cell protein and cell carbohydrate levels of the flagellatePolytomella agilis during growth in batch cultures on a complex medium at 25° C, 18° C and 9° C were examined. At 25° C, cell protein and carbohydrate levels fell markedly during exponential population growth. At 18° C, cell protein values remained fairly constant, while cell carbohydrate increased. During growth at 9° C, both cell protein and carbohydrate increased. Changes in these cellular parameters were related to marked differences in the rates of population growth at these temperatures. The cellular locus of changes observed at 25° C was examined by centrifugal fractionation of disrupted cells on density gradients.Encystment was studied in cultures grown at 25° C. The maximum number of cysts produced was 5–10 per cent of the peak population density; the rate and degree of encystment were not increased by growth on conditioned medium or starvation. Cysts isolated from mixed populations by a centrifugal procedure remained viable, even after storage at —5° C for several weeks. The soluble proteins of separately disrupted motile and encysted forms ofP. agilis were examined and compared by disc electrophoresis.This investigation was supported by U.S.P.H.S. Grant AI-07926-02.     

Limitation of tobacco callus tissue growth by carbohydrate availability

Growth rate, maximum dry weight yield, and carbohydrate utilization were measured with pith callus derived from Nicotiana tabacum L. var. Wisconsin No. 38. Maximum tissue dry weights increased as carbohydrate (either glucose or sucrose) in the medium was increased. The time at which maximum growth was obtained coincided with the time at which carbohydrate was exhausted from the medium. The addition of carbohydrate to the medium before the end of log phase growth supported that amount of additional growth which would have been obtained if all of the carbohydrate had been added to the medium prior to planting the tissue. Maximum obtainable dry weights at logarithmic growth rates greater than 0.16 doubling per day depended on the amount of carbohydrate in the medium and not on a particular hormonal regime.     

Culture conditions for efficient induction of green plants from isolated microspores of barley

Subculture regime and carbohydrate concentration of the medium had a marked effect on the regeneration of green plantlets from mechanically isolated microspores of Hordeum vulgare L. cv. Kymppi. A sevenfold increase in the yield of green plants was obtained by shortening the suspension culture time of the developing proembryo mass from 4 to 3 weeks. A further twofold increase was obtained by increasing the maltose concentration of the microspore isolation medium and of the culture medium. Under optimal conditions, a mean of 169±97 green plants per spike were regenerated.     

Carbohydrate supplements to the callus culture medium modify the growth of potato tuber moth larvae feeding on Lycopersicon chmielewskii callus

Lycopersicon esculentum and L. chmielewskii are respectively susceptible and resistant to the potato tuber moth (Phthorimaea operculella Zeller) in the field. Feeding bioassays were conducted with the herbivore caterpillars reared on callus derived from both tomato species and grown in vitro, and the influence of carbohydrate supplements to the callus culture medium, on the insect's feeding behavior was investigated. Newly-hatched larvae fed with L. esculentum or L. chmielewskii callus raised on a medium with 88 mM sucrose, reached a weight of 12–15 mg and 1.5–3.0 mg, respectively, within 9 days. Restriction of larval weight increase in insects reared on L. chmielewskii callus, disappeared when the host tissue was transferred 24 h prior to the callus-insect assay to a medium supplemented with 264 mM of either sucrose, glucose, fructose or mannose. The capability of L. chmielewskii callus to restrict growth of larvae was restored in host tissue retransferred from a medium with 264 mM sucrose to a 24-h incubation on one supplemented with 264 mM of either mannitol, sorbitol, glycerol or myo-inositol, before the callus-insect bioassay. The larval growth response remained unaltered by callus incubated on a medium with 264 mM xylose. The ameliorating effect on insect growth of high sucrose in the callus medium was not due to sucrose as an ingredient of the insect's diet. The diverse response of L. chmielewskii callus, and its dependence on the type of carbohydrate in the medium, rule out effects of these substances as nonspecific medium osmotica. The swift callus responses to carbohydrates (within hours of a change in medium composition), as reflected in the insect's growth, were not accompanied by visible morphological variations in the host tissue. We suggest that suppression by high levels of exogenously applied saccharides and derepression by exogenous polyols and myo-inositol of the impedement to growth of the potato tuber moth larva, reflect the existence in L. chmielewskii of a carbon metabolic control mechanism of gene expression whose products affect insect growth.