Photo-Assimilation of Acetate by an Obligate Phototrophic Strain of Euglena gracilis

SYNOPSIS. Light-dependent incorporation of acetate occurs in an obligate phototrophic strain of Euglena gracilis (strain L). Assimilation is into all major biochemical fractions. Acetate does not induce operation of the glyoxylate by-pass as it does in heterotrophic strains; neither does it stimulate oxygen consumption. Acetate will not replace CO₂ in phototrophic growth. A number of carbon sources tested would not support growth in the dark, and glucose was not incorporated either in the light or the dark.     

NUTRITION OF PANDORINA MORUM1,2

A defined medium was developed for 3 strains of Pandorina morum. The strains tested required no vitamins or other organic compounds. The optimal initial pH was between 7.0 and 8.0. Various carbon sources were tested, and only glycolate and acetate appreciably stimulated growth. Mixotrophic growth in the light was stimulated by glycolate in all 3 strains, and by acetate in strains 880 and N76-6. Only strain N76-6 utilized acetate for heterotrophic growth in the dark. Thirty strains of P. morum of world-wide distribution were surveyed for mixotrophic and heterotrophic growth with acetate. All were found to fit 1 of 3 classes with respect to acetate metabolism: (1) no effect in light or dark; (2) stimulation of growth in light only; (3) stimulation of growth in light and dark.     

GROWTH,PHYSIOLOGY, AND ULTRASTRUCTURE OF A DIAZOTROPHIC CYANOBACTERIUM,CYANOTHECE SP. STRAIN ATCC 51142, IN MIXOTROPHIC AND CHEMOHETEROTROPHIC CULTURES1

The growth, physiology, and ultrastructure of the marine, unicellular, diazotrophic cyanobacterium, Cyanothece sp. strain ATCC 51142, was examined under mixotrophic and chemoheterotrophic conditions. Several organic substrates were tested for the capacity to support heterotrophic growth. Glycerol was the only substrate capable of enhancing mixotrophic growth in the light and supporting chemoheterotrophic growth in the dark. Dextrose enhanced mixotrophic growth but could not support chemoheterotrophic growth. Chemoheterotrophic cultures in continuous darkness grew faster and to higher densities than photoautotrophic cultures, thus demonstrating the great respiratory capacity of this cyanobacterial strain. Only small differences in the pigment content and ultrastructure of the heterotrophic strains were observed in comparison to photoautotrophic control strains. The chemoheterotrophic strain grown in continuous darkness and the mixotrophic strain grown in light/dark cycles exhibited daily metabolic oscillations in N₂ fixation and glycogen accumulation similar to those manifested in photoautotrophic cultures grown in light/dark cycles or continuous light. This “temporal separation” helps protect O₂-sensitive N₂ fixation from photosynthetic O₂ evolution. The rationale for cyclic glycogen accumulation in cultures with an ample source of organic carbon substrate is unclear, but the observation of similar daily rhythmicities in cultures grown in light/dark cycles, continuous light, and continuous dark suggests an underlying circadian mechanism.     

Physiological characteristics of cyanobacteria in pulp and paper waste-treatment systems

An investigation was made into the physiological characteristics of the extensive cyanobacterial communities in pulp and paper secondary waste-treatment systems, including the capacity of isolates to biodegrade organic contaminants in these systems. Although pulp and paper waste-treatment systems were found to be severely light-limited, photosynthesis-irradiance curves indicated that shade-adapted cyanobacterial communities could fix conspicuous amounts of inorganic carbon via photosynthesis. Of 21 cyanobacterial strains isolated from pulp and paper waste-treatment systems located in 4 countries, all except one were capable of glucose uptake in the light, and 19 also showed uptake in the dark. In the 6 species tested, glucose and acetate addition stimulated growth in low light in all except one species. Aphanocapsa rivularis grew equally well on glucose and acetate in the dark but Pseudanabaena sp. grew well on acetate, and minimally on glucose. Growth stimulation by glucose and acetate in these two strains was greater in low than in high light. Pseudanabaena sp. and Phormidium animale both accumulated 2,4-dichlorophenol and 3-chlorobenzoate but showed minimal mineralization to CO₂. None of the four species tested could accumulate or degrade phenol or dichloroacetate. It is concluded that, depending on the light conditions, cyanobacteria contribute organic carbon in photosynthesis, and/or remove small organic molecules during mixotrophic and heterotrophic growth but are not important degraders of contaminants in these waste treatment systems.     

HETEROTROPHIC GROWTH OF ULVA LACTUCA (CHLOROPHYCEAE)1

The effect of external glucose (51 mM) and acetate (13 mM) on growth and photosynthetic capacity of Ulva lactuca L. was tested in laboratory cultures over 41 days in the dark and in dim light (0.9 μmol photons·m^?2·s^?1) at 7–8° C. Glucose and acetate had a significant positive effect on growth rate, chlorophyll content, and quantum yield for discs grown in the dark and in dim light. The carbon gain from heterotrophic uptake was low and only allowed U. lactuca to maintain a specific uptake was low and only allowed U. lactuca to maintain a specific growth rate of 0.005 day^?1 compared to 0.06–0.1 day^?1 at higher light intensities. However, plants with added organic substrate maintained a normal chlorophyll content and were able to photosynthesize whereas control plants lost pigmentation and photosynthetic capability after 41 days in both dim light and darkness, probably because of disorganization of the photosynthetic apparatus. This suggest that the ecological significance of heterotrophic uptake is to allow U. lactuca to survive during prolonged low light conditions with an intact photosynthetic apparatus.     

Differences between strains of rhizobium in sensitivity to canavanine

Summary Four strains of rhizobia that nodulate canavanine-synthesizing legumes and four strains that nodulate noncanavanine-synthesizing legumes were tested for sensitivity to L-canavanine. The effect of canavanine on growth depends upon the strain of Rhizobium tested rather than the canavanine synthesizing capability of the host legume. In both groups of rhizobia, some strains were inhibited in growth by canavanine. Canavanine enhancement of growth was observed in rhizobia that nodulate noncanavanine-synthesizing legumes.Canavanine was found to enhance incorporation of uridine-H³ and L-leucine-H³ into trichloroacetic acid insoluble fractions of starved cells of two strains of rhizobia tested. This demonstrated that under certain conditions some rhizobia can detoxify canavanine and utilize it in synthetic processes. re]19760729     

Fissazione di C14O2 in Colture di Tessuti Vegetali « in Vitro »

Abstract

C¹⁴O₂ fixation in plant tissues « in vitro ». — In the present work it has been examinated the autotrophic and heterotrophic CO₂ fixation of explants of « Helianthus tuberosus » « in vitro » and the photosyntetic efficiency of leaves produced from buds of « in vitro » explants of « Cichorium intybus » compared with that of mature leaves from normal plants of the same species. From our results it is evident that « in vitro » explants of « Helianthus tuberosus », grown, in the light, are able to autotrophically incorporate C¹⁴O₂; the distribution of the radioactivity into the various fractions shows a large influence of the light on the neutral fraction containing sugars (50% of the total radioactivity). In the chlorophyllous explants the dark CO₂ fixation is obviously of heterotrophic type: 97% of the total radioactivity is incorporated in amine acids (43%) and the organic acids (53%); on the other hand in the dark grown explants the radioactivity is differently distributed between amino acids (59%) and organic acids (39%). Mature leaves from normal plants and leaves produced from buds of « in vitro » explants of « Cichorium intybus » incorporate the same quantity of C¹⁴O₂ when expressed per mg of chlorophyll; the different distribution of the radioactivity in the neutral and acid fractions could be explained in terms of a different utilization pathway of the photosynthates in the two tissues.     

Studies on the biosynthesis of protein and lipid components of rat liver mitochondria

1. Male rats were injected intraperitoneally with l-[³⁵S]methionine, [³²P]-phosphate and [2-¹⁴C]acetate. The animals were killed at various times up to 72hr. after injection, and liver mitochondria were prepared and fractionated into soluble protein, insoluble protein and lipid for assay of the radioactivity of each fraction. 2. The maximal specific radioactivity of total mitochondrial phospholipid with respect to both ³²P and ¹⁴C was attained after approx. 6hr. 3. ³²P was incorporated most rapidly into phosphatidylethanolamine, maximal incorporation being attained after approx. 6hr.; maximal incorporation into lecithin occurred after 6–12hr. The specific radioactivity of cardiolipin was still slowly increasing at the end of the experiment (72hr.). 4. There were no major differences between the rates of incorporation of ¹⁴C into the lecithin, phosphatidylethanolamine and cardiolipin fractions of mitochondrial phospholipid, maximal incorporation in each case occurring after approx. 6hr. 5. Maximal incorporation of ³⁵S into both soluble and insoluble protein fractions was attained less than 12hr. after injection, the maximal specific radioactivity of soluble protein being higher than that of insoluble protein.     

Studies on Orobanche hederae Physiology: Pigments and CO2 Fixation

In order to investigate some aspects of Orobanche hederae physiology in relation to its parasitism, the pigment composition and the ¹⁴CO₂ incorporation, both in light and in dark, were studied. By means of various chromatographic techniques, it was shown that chlorophyll is probably quite absent and that the carotenoids are very largely distrihuted and consist almost exclusively of xanthophylls. Flavochrome, flavoxanthin, and a pigment resembling neoxanthin were found to be the major components; lutein-5,6-epoxide, taraxanthin, and traces of β-carotene and α-carotene-5,6-epoxide were also present. CO₂ incorporation is completely of heterotrophic type, being in no way stimulated by the light; the major radioactivity was detected in the fractions containing amino acids and organic acids.     

ACETATE IS A CHEMOATTRACTANT FOR THE COLONIAL GREEN ALGA ASTREPHOMENE GUBERNACULIFERA (CHLOROPHYCEAE)1

Astrephomene gubernaculifera Pocock is capable of growth on sodium acetate, even in the absence of light. From an evolutionary standpoint, it would be beneficial for this motile heterotrophic organism to be able to sense and swim to such an energy source. We used three assays to test for chemoattraction of A. gubernaculifera. In each case, there was strong chemoattraction to acetate. Propionate, a molecule structurally similar to acetate, also elicited a strong chemoattraction response, even though it was incapable of supporting growth in the dark in this strain. This implies that chemoattraction in A. gubernaculifera is based on a receptor‐mediated signaling response. Video analysis of colonies exhibiting the attraction response indicated that most of them remained motile in the vicinity of the acetate. This rules out the simplest possible model for accumulation in which colonies stop when they encounter increased acetate concentrations. We also tested a variety of other organic molecules for their ability to chemoattract A. gubernaculifera. Of the 11 sugars, 18 amino acids, and three other organic molecules tested, only the amino acid phenylalanine elicited chemoattraction.     

PRECURSOR INCORPORATION INTO CORTICAL PROTEIN DURING FIRST EXPOSURE OF RATS TO LIGHT; CELLULAR LOCALIZATION OF EFFECTS

—The rate of incorporation of [³H]lysine into acid-insoluble material in vivo was determined in neurons and neuropil from the visual cortex of dark-reared rats, littermates exposed to the light for varying lengths of time and normally reared controls. Following onset of light exposure, the elevation of incorporation was confined to the neuronal fraction. On continuous exposure for up to 96 h, the level of incorporation in the neuronal fraction dropped to that of the dark control value. In dark-reared animals, the rate of incorporation in the neuronal fraction was 68 per cent of that in neuropil, in normals it was 150 per cent. On onset of exposure, the ratio in light exposed animals approached the normal level, but on prolonged continuous exposure both light exposed and normal ratios dropped to the dark control value once more. This drop did not occur if the animals were exposed to a 12 h light/dark cycle. These results are taken as suggesting that part of the protein synthesis of the visual cortex is functionally controlled, and that neuronal and neuropil fractions show a metabolic relationship which can be affected by environmental changes. The failure to show a depression of incorporation in prolonged exposure, by comparison with earlier results under somewhat different behavioural conditions, was taken as further evidence for the ‘state-dependence’ of a number of brain biochemical parameters.     

Trophic dependencies of some larval chironomidae (Diptera) and fish species in the River Thames

Experiments are described to characterise the heterotrophic potential of Westiellopsis prolifica Janet, which fixes nitrogen under light and dark conditions. The growth of the organism in terms of dry weight increase, was more in fructose, lactose, sucrose, sorbose, galactose, glucose, sodium acetate, mannitol, sorbitol, glycerol, ethyl alcohol and butyl alcohol, when the alga was pretreated with light and subsequently incubated with the substrates in light. Mannose, xylose, acetic acid, propionic acid, fructose 1,6 di Po₄, pyruvic acid, dihydroxyacetone and succinic acid decreased the growth of the organism in the same condition. In dark incubation after pretreatment with light, as well as in the dark, Westiellopsis showed a better growth response to almost all the exogenous substrates. However, after pretreatment either with light or dark, the test organism utilised exogenous substrates quicker in light than in dark incubations. These experiments would suggest that the substrate specificity and efficiency of substrate utilisation by the alga during its heterotrophic growth are governed by the growth conditions.     

Effects of prey abundance and light intensity on the mixotrophic chrysophyte Poterioochromonas malhamensis from a mesotrophic lake

1. Previous studies of mixotrophy in the flagellate Poterioochromonas malhamensis (Chrysophyceae) were performed on strains that had been in culture for > 30 years. This study aims to compare mixotrophy in a cultured strain with one recently isolated from a mesotrophic lake (Lacawac) in Pennsylvania, U.S.A. 2. P. malhamensis from the lake exhibited a nutritional flexibility similar to that of the culture strain, growing phototrophically but inefficiently in comparison to other nutritional modes (growth rate (μ) = 0.015 h^?1). Supplementing an inorganic salts medium with 1 mM glucose resulted in a doubling of μ to 0.035 h^?1 and 0.033 h^?1 in the light and the dark, respectively. Addition of an algal prey, Nannochloris, to the inorganic salts medium increased growth to rates similar to those observed with glucose. Maximum growth of the lake strain, 0.095 h^?1, was achieved when bacteria was supplied as food. During growth on bacteria, cellular chlorophyll a (Chl a) decreased from 140 fg cell^?1 to 10 fg cell^?1 over 22 h when cultured either in the light or dark. In illuminated cultures, cell-specific Chl a concentration recovered to 185 fg cell^?1 after bacteria became limiting. 3. In contrast to the cultured strain, however, the lake isolate exhibited an inverse relationship between light intensity and ingestion rate. Calculated grazing rates, based upon the ingestion of fluorescently labeled bacteria, were 3.2, 5.2 and 9.4 bacteria flagellate^?1 h^?1, for P. malhamensis incubated in high light, low light and darkness, respectively. Phagotrophy is thus influenced by a light regime in this predominately heterotrophic mixotroph.     

Polyribonucleotide synthesis by subfractions of microsomes from rat liver

1. The microsome fraction of rat liver has been fractionated and the ability of the fractions to incorporate ribonucleotides into polyribonucleotides has been studied. Activity was found in the rough-surfaced vesicle (light) fraction and in the free-ribosome fraction and this latter activity has been examined. 2. The free-ribosome fraction contains ribosome monomers, dimers and trimers together with some higher oligomers and ferritin. In addition to catalysing the incorporation of ribonucleotides into acid-insoluble material it contains diesterase activity. It catalyses the incorporation of UMP from UTP, but not UDP, AMP from ATP and CMP from CTP into polyribonucleotide material, and for UTP the product appears to be a homopolymer not more than eight units long attached to the ends of primer polyribonucleotide strands. 3. The activity could not be removed from the free-ribosome fraction by washing or by isolation in the presence of ethylenediaminetetra-acetic acid. 4. Partially hydrolysed polyuridylic acid but not polyadenylic acid could serve as a primer for the incorporation of UMP, but some activity was always associated with an endogenous primer. 5. Analysis of RNA extracted from the free-ribosome fraction after incubation with [³H]UTP showed the presence of 28s, 18s, 5s and transfer RNA types, but no radioactivity was associated with any of these RNA fractions.     

THE UPTAKE OF MANNITOL AND SORBITOL BY A SPECIES OF CHLORELLA (CHLOROPHYCEAE)1

Chlorella saccharophila (Krüger) Nadson takes up mannitol and sorbitol in the light and the dark. The rate of uptake is concentration dependent. is not affected by pH in the range pH 6.0 to 8.0 and ii not stimulated by light. Uptake is inhibited by the respiration inhibitor sodium azide (10^-2 M) but not by 3-(3,4-dichlorophenyl)-1,1-di-methyl urea (10^-6 M), an inhibitor of photosynthesis. Sorbitol. but not mannitol, stimulates the rate of dark respiration but both support the heterotrophic growth of the alga. Both compounds permeate the cells of C. miniata. and two strains of C. pyrenoidosa but do not support the heterotrophic growth of these algae. The cells of C. vulgaris are impermeable to both compounds.     

Synthesis of DNA during the cell cycle of synchronous Anacystis nidulans

Cells of Anacystis nidulans strain 1402-1 incorporate [methyl-³H]thymidine or [8-³H]adenine into DNA; in synchronous cultures (21/2 h full light, 1/2 h weak light, 5 h dark), this incorporation occurs in the dark to different extents according to the labeled precursor offered or to its specific activity. The specific activity of in vivo, uniformly labeled DNA decreases to half the initial value when the cells are grown in the absence of radioactive DNA precursors during the light phase; it does not decrease during the following dark phase. If unlabeled thymidine is given during the dark phase, the specific activity of the DNA starts to decrease at the onset of the next light phase. The time course of the decrease supports the hypothesis that all cells start their DNA replication immediately after illumination and that the first cells have completed if after 1.25 h. The slowest cells then need 3.75 h for completion of DNA replication. It is discussed whether the incorporation during the dark might be due to pool size effects.     

The Relationship between Growth and Proline Incorporation after Auxin Treatment of Mung Bean Hypocotyls

Indoleacetic acid (IAA) stimulates the incorporation of ¹⁴C-proline into both the cyloplasmic and the cell wall fractions of the hypocotyl of mung bean (Phaseolus aureus Roxb. cv. Black). It neither stimulates the transfer of ¹⁴C-proline from the cyloplasmic fraction into the cell wall fraction, nor the retention of ¹⁴C-proline in the wall or cytoplasmic fractions. Moreover, the stimulation of growth caused by IAA parallels the stimulation of the incorporation of proline into the cytoplasmic fraction, but does not parallel the stimulation into the cell wall fractions. The stimulation of the incorporation into the cyloplasmic fraction seems to appear within 30 minutes after auxin treatment, at about the same time the increase in the growth is observed in response to IAA, suggesting a connection between these effects. On the other hand, the stimulation of the proline incorporation into the cell wall fraction seems to require more than 90 minutes after auxin treatment, suggesting no close connection between growth and proline incorporation into the cell wall fraction.     

Environmental factors affectingin vitro nitrogenase activity of cyanobacteria isolated from rice-fields

The nitrogen-fixing capacity of four cyanobacterial strains was tested in relation to heterotrophic ability, tolerance to combined nitrogen and adaptive capacity to changes in light intensity and pH. Strains (Anabaena variabilis UAM 202;Calothrix marchica UAM 214;Nodularia spumigena UAM 204,Nostoc punctiforme UAM 205) were isolated from the rice-fields of Valencia (Spain).C. marchica, was the only strain able to grow and to fix dinitrogen under heterotrophic conditions, with fructose and glucose. Fructose was the best substrate supporting growth and dinitrogen fixation in mixotrophy (growth in the light under conditions where CO₂ and organic carbon are assimilated simultaneously), photoheterotrophy (growth in the light on an organic compound in the absence of net CO₂ fixation) and heterotrophy (growth on an organic compound in the dark). Ammonium repressed nitrogenase more than nitrate. Full repression was observed only at concentrations of combined nitrogen higher than those usually found in rice-fields. Carbohydrates had a protective effect on nitrogenase against ammonium inhibition inC. marchica. All four strains showed increased nitrogenase activity when the light intensity was increased during assays. Variations of pH normally occurring in rice fields led to no important changes in nitrogenase activity inC. marchica. This fact, together with its potential for heterotrophic growth and tolerance to combined nitrogen, make this the most suitable of the four strains for inoculation experiments in rice fields.     

Nutritional characteristics of a mixotrophic nanoflagellate,Ochromonas sp.

Autotrophic and heterotrophic growth characteristics of a nano-flagellate were investigated. The flagellate,Ochromonas sp., was isolated from the northern Baltic Sea. Autotrophic growth was poor. Axenically pregrown flagellates did not increase significantly in cell number during incubation in different inorganic media. The number of flagellates remained constant 3–5 weeks in cultures kept in the light (100mol m^–2 sec^–1), whereas in the dark, a high mortality rate was found. Uptake of inorganic¹⁴C into an acid-stable fraction indicated thatOchromonas had a functional photosynthetic apparatus. Heterotrophic growth in both liquid medium and medium containing bacteria was rapid. The maximum growth rate corresponded to a generation time of 5.3 hours. Light had no effect on heterotrophic growth. Cells pregrown onEscherichia coli minicells survived without additional bacteria as food when kept in the light, but rapid death occurred in darkness. In conclusion, heterotrophy is the major mechanism to support growth in this species ofOchromonas, but under poor environmental conditions photoautotrophy might be a strategy for survival rather than growth.