Pseudomonas putida A ATCC 12633 oxidizes trimethylamine aerobically via two different pathways

The present study examined the aerobic metabolism of trimethylamine in Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine. In both conditions, the trimethylamine was used as a nitrogen source and also accumulated in the cell, slowing the bacterial growth. Decreased bacterial growth was counteracted by the addition of AlCl₃. Cell-free extracts prepared from cells grown aerobically on tetradecyltrimethylammonium bromide exhibited trimethylamine monooxygenase activity that produced trimethylamine N-oxide and trimethylamine N-oxide demethylase activity that produced dimethylamine. Cell-free extracts from cells grown on trimethylamine exhibited trimethylamine dehydrogenase activity that produced dimethylamine, which was oxidized to methanal and methylamine by dimethylamine dehydrogenase. These results show that this bacterial strain uses two enzymes to initiate the oxidation of trimethylamine in aerobic conditions. The apparent K_m for trimethylamine was 0.7 mM for trimethylamine monooxygenase and 4.0 mM for trimethylamine dehydrogenase, but both enzymes maintain similar catalytic efficiency (0.5 and 0.4, respectively). Trimethylamine dehydrogenase was inhibited by trimethylamine from 1 mM. Therefore, the accumulation of trimethylamine inside Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine may be due to the low catalytic efficiency of trimethylamine monooxygenase and trimethylamine dehydrogenase.     

Effect of Ethidium Bromide on the Oxidative Metabolism and Enzyme Profiles of Crithidia fasciculata*

SYNOPSIS. Changes in the metabolism of Crithidia fasciculata ATCC 11745 when grown in the presence of ethidium bromide were studied. Ethidium bromide-grown cells had decreased respiratory activity as measured by oxygen consumption. More than 50% of the organisms cultivated in a defined medium containing 1.0 mg/liter of ethidium bromide became dyskine-toplastic and had decreased activities of particulate succinate and NADH-linked dehydrogenases as well as of soluble isocitrate dehydrogenase. These cells also had increased activities of particulate α-glycerophosphate dehydrogenase, soluble α-glycerophosphate dehydrogenase, malic enzyme, hexokinase, and malate dehydrogenase. Ethidium bromide-grown cells had a lower level of ATP and contained less DNA than cells grown in its absence.     

Field experiments in the control of Verticillium dahliae and Heterodera rostochiensis on potatoes

Potatoes were grown in soil infested with Verticillium dahliae and Heterodera rostochiensis and treated with methyl bromide, aldicarb or benomyl. Successive crops were grown in subsequent years on the same plots but without further treatment. Largest yields in the year of treatment were from plots fumigated with methyl bromide but in the second crop benomyl-treated plots yielded most. Neither methyl bromide nor aldicarb affected yields from trie third crop. After lifting the first crop, soil from plots treated with methyl bromide or benomyl contained less V. dahliae than that from plots receiving aldicarb or nothing. Soil was also less infective after the second crop following methyl bromide fumigation, but not the third. Plots treated with methyl bromide or aldicarb contained many fewer H. rostochiensis than untreated plots after the first crop, slightly less after the second and equal numbers after the third crop. H. rostochiensis were also fewer in benomyl-treated plots than in untreated after the first crop, but, surprisingly, were fewest after the second crop. The increased yield after applying benomyl to soil seems to depend more on its effect on H. rostochiensis than on V. dahliae. Fumigating with methyl bromide decreased common scab and black scurf on progeny tubers but increased infections by Oospora pustulans.     

Fine structure of the epidermis in Gnathostomulida

Summary The fine structure of the epidermis in Haplognathia simplex, Haplognathia rosea, Pterognathia meixneri (Filospermoidea) and Gnathostomula paradoxa (Bursovaginoidea) has been investigated. The epidermis in the filospermoidean species is uniform, consisting of epidermal cells with a single locomotory cilium. The structure and development, including ciliogenesis, of these epidermal cells are described. In G. paradoxa additional epidermal elements have been found: mucous cells with a presumably apocrine secretion modus are scattered in a strip-like arrangement within the epidermis. Their deverlopment is separate from epidermal cells with locomotory function. Two further types of glandular cells with either a single cilium or a diplosome are located ventrally. It is assumed that they represent an adhesive system.Abbreviations (used in figures) ac accessory centriole - ap appendix of accessory centriole - ax axoneme - bb basal body - bf basal foot - bl basal lamina - c cilium - cA ciliary adhesive cell - ce centriole - cp ciliary pit - d diplosome - dy dictyosome - dA diplosomal adhesive cell - E epidermal cell with locomotory cilium, epidermis - ev epidermal vesicle (epitheliosome) - gv gland vesicle - m mitochondrion - ma microvillus of apical cell membrane - mp microvillus of ciliary pit - mv microvillus - n nucleus - ps prosecretory-vesicle - R receptor - r ciliary rootlet - rc caudal ciliary rootlet - rr rostral ciliary rootlert - sv secretory vesicle - v vesicle - v _i central vesicle of multivesicular body - v _o surrounding vesicle of multivesicular body - z cisternae     

Aerobiosis and the hemoprotein content of photosynthetic bacteria

Summary Rhodospirillum rubrum and Rhodopseudomonas spheroides, grown under various degrees of illumination, aeration, and iron deprivation, have been assayed for their content of cytochrome c, RHP, catalase, total iron, bacteriochlorophyll, and carotenoids.Concentrations of bacteriochlorophyll and carotenoids were consistent with the findings of Cohen-Bazire et al. (1957).Total iron content, which ranged from 0.017 to 0.04% of the dry weight, reflected the content of the principal hemoproteins but exceeded the amount of iron in these hemoproteins.The catalase content of R. rubrum, on a dry weight basis, was 0.0005% for cells grown anaerobically in the light, and 0.0028% for cells grown in darkness with vigorous aeration; that of Rps. spheroides was 0.006% and 0.25%, respectively. The catalase content in both species rose with increasingly vigorous aeration.Cytochrome c in both species, and RHP in R. rubrum, attained the same levels in cells grown under vigorous aeration as in cells grown anaerobically in the light. In cells grown under limited aeration the levels of these substances were about 50% higher. In Rps. spheroides the RHP content was greatest in cells grown anaerobically, falling under gentle aeration and declining further under more vigorous aeration.Iron deficiency caused a decrease in the catalase content of cells grown anaerobically in the light but not in cells grown aerobically. The content of cytochrome c and of RHP was diminished by iron depletion in aerobic cultures, but not in anaerobic cultures.operated by Union Carbide Corporation for the U.S.Atomic Rnergy Commission.     

Abstracts of Papers Read at the Winter Meeting at the University of Liverpool

Previous investigations with planktonic cyanobacteria have suggested that these organisms do not form new gas vesicles in the dark. This study, on Microcystis sp., confirmed that cells that had been preincubated at low photon irradiances (< 15 μmol m^-2 s^-1) formed negligible amounts of gas vesicles in the dark. Significant gas vesicle formation occurred, however, in cells preincubated continuously at higher irradiances, and particularly within the range 65 to 105 μmol m^-2 s^-1. The results suggest that gas vesicle formation in the dark is dependent on the prior accumulation of energy reserves. The amount of gas vesicles formed in continuous light was linearly related to irradiance over the range 0 to 20 μmol m^-2 s^-1, and reached a maximum at only 30 μmol m^-2 s^-1 that was over five times the amount formed at higher irradiances. This suggests that the rate of gas vesicle formation, regulated directly in response to irradiance, has a role in the light-mediated buoyancy regulation of this cyanobacterium.     

The ultrastructure of the anal vesicle of the parasitoid,Microplitis croceipes (Hymenoptera:Braconidae)

The fine structural characteristics of epithelial cells of the anal vesicle in the hymenopteran parasitoid, Microplitis croceipes (Cresson), are similar to those of transport cells. Apical and basal infoldings, an abundance of mitochondria, ribosomes, rough endoplasmic reticulum, Golgi complexes and pinocytotic vesicles all indicate a transport function for these epithelial cells. The medial portions of both Malpighian tubules located within the anal vesicle also were examined and on the basis of morphology appear to be active. These observations support earlier physiological data which indicate that the anal vesicle functions in absorption of nutrients and excretion.     

Influence of ethidium bromide on respiration in Schizosaccharomyces pombe

Summary In Schizosaccharomyces pombe ethidium bromide inhibits respiratory competence in all treated cells. After inoculation in ethidium bromide free medium these cells show reversion to respiration competence.     

Effect of aspirin on mitochondrial mutagens in Saccharomyces cerevisiae

Summary The mitochondrial mutation petite was induced in yeast cells by ethidium bromide (EB), Adriamycin (ADR) and 4-nitroquinoline-N-oxide (NQO). In the presence of aspirin in concentrations ranging from 0.1 to 1.0 mg/ml, the mutagenicity of EB and ADR was reversed but petite induction by NQO was unaffected. At these concentrations, aspirin also reversed mitochondrial inhibition by oligomycin, a non-mutagenic inhibitor of the organellar ATPase complex.Cells grown in the presence of aspirin alone showed a significantly higher rate of oxygen uptake than untreated control cultures when the drug concentration ranged from 0.05 to 1.0 mg/ml. At concentrations of 2 mg/ml and above, aspirin inhibited mitochondrial respiration.     

An ultrastructural study of the polyp and strobila of Atorella japonica (Cnidaria,Coronatae) with respect to muscles and nerves

Atorella japonica were observed by TEM to examine the nerve plexus in the capitulum of the polyp and the cross-striated muscle cells of the strobila. The nerve plexus included a number of neuromuscular junctions and many interneural synapses. Neuromuscular junctions contained two types of synaptic vesicle: clear and small (ca 75 nm diam.), and dense cored and large (ca 120 nm diam.). The first type of vesicle always appeared near the presynaptic membrane and the second type was distributed behind the former. In interneural synapses, two types of vesicle which were similar to neuromuscular synaptic vesicles were recognized. They were distributed in a pattern similar to that of the neuromuscular synaptic vesicles, but these vesicles were found on both sides of the two synaptic membranes.     

Oogenesis of microlecithal oocytes in the viviparous teleost Heterandria formosa

Viviparous teleosts exhibit two patterns of embryonic nutrition: lecithotrophy (when nutrients are derived from yolk that is deposited in the oocyte during oogenesis) and matrotrophy (when nutrients are derived from the maternal blood stream during gestation). Nutrients contained in oocytes of matrotrophic species are not sufficient to support embryonic development until term. The smallest oocytes formed among the viviparous poeciliid fish occur in the least killifish, Heterandria formosa, these having diameters of only 400 μm. Accordingly, H. formosa presents the highest level of matrotrophy among poeciliids. This study provides histological details occurring during development of its microlecithal oocytes. Five stages occur during oogenesis: oogonial proliferation, chromatin nucleolus, primary growth (previtellogenesis), secondary growth (vitellogenesis), and oocyte maturation. H. formosa, as in all viviparous poeciliids, has intrafollicular fertilization and gestation. Therefore, there is no ovulation stage. The full‐grown oocyte of H. formosa contains a large oil globule, which occupies most of the cell volume. The oocyte periphery contains the germinal vesicle, and ooplasm that includes cortical alveoli, small oil droplets and only a few yolk globules. The follicular cell layer is initially composed of a single layer of squamous cells during early previtellogenesis, but these become columnar during early vitellogenesis. They are pseudostratified during late vitellogenesis and reduce their height becoming almost squamous in full‐grown oocytes. The microlecithal oocytes of H. formosa represent an extreme in fish oogenesis typified by scarce yolk deposition, a characteristic directly related to matrotrophy. J. Morphol., 2011. © 2010 Wiley‐Liss, Inc.     

Observations on the origin of two substances produced by the vitellaria in Gorgoderina attenuata

The vitelline cells in Gorgoderina attenuata produce two qualitatively distinct substances. One substance assumes the form of individual, dense, osmiophilic globules. Many globules are contained in a single vesicle. The other substance is an amorphous mass of medium density that completely fills a vesicle. Observations indicate that the dense, osmiophilic globules develop in association with a system of small, contiguous, ribosome-free vesicles. It is suggested that this system of vesicles constitute a Golgi apparatus for these cells. The amorphous mass substance develops in vesicles which appear to be derived from endoplasmic reticulum. Close association between the amorphous mass vesicle and mitochondria are commonly observed.     

Algal affinity and possible life cycle of the early Cambrian acritarch Yurtusia uniformis from South China

Abundant, well-preserved specimens of spheroidal organic-walled microfossil Yurtusia uniformis are reported from the basal Cambrian Yanjiahe Formation in the Changyang area of Hubei Province, South China. Thin and hollow processes extend between the double walls of the vesicle. The single to multiple internal bodies within the vesicle cavity are observed in the genus for the first time, representing reproductive structures (dividing daughter cells). A small circular perforation may occur on the vesicle wall to release the internal bodies. Morphological analyses of specimens preserved at various life stages reveal that processes gradually became longer as the vesicle grew in size. The internal bodies (daughter cells) underwent several successive divisions within the vesicle, which was accompanied by the simultaneous growth of both vesicle and processes. The regular growth of cells, formation and release of daughter cells, and the remarkable morphological similarity between extant algae and the studied microfossils suggest that Yurtusia uniformis is probably a green microalga that may be closely related to the Trebouxiophyceae or even Chlorellales (Chlorophyta). The growth and reproductive mode of individuals indicates that Y. uniformis is an actively growing vegetative cell of microalgae, rather than a metabolically inert cyst or resting spore. A life cycle involving vegetative growth and asexual reproduction is proposed for Y. uniformis on the basis of the life histories of modern chlorophytes. The multiple internal cells may represent autospores produced by a mature autosporangium during asexual reproduction, which subsequently developed into separate young vegetative cells after their release from the opened autosporangium.     

Growth of Candida albicans in the presence of hydrocarbons: a correlation between sterol concentration and hydrocarbon uptake

Summary Candida albicans KTCC 89062 grown on n-alkanes showed higher levels of sterol content as compared to glucose-grown cells. Certain sterols, such as lanosterol, were significantly reduced in cells grown on n-alkanes, while others, such as ergosterol, increased in these cells. Sterol fractions declined as the chain length of the n-alkanes increased. Ergosterol supplementation of the chemically defined medium showed an increase in the uptake of dodecane (C₁₂) by cells grown on such medium. Increase in the concentration of ergosterol supplementation resulted in an increase in C₁₂ uptake. The uptake of C₁₂ was not stimulated by ergosterol supplementation in the case of non-viable yeast cells.     

Ocelli in a Cnidaria polyp: the ultrastructure of the pigment spots in Stylocoronella riedli (Scyphozoa,Stauromedusae)

Within the Cnidaria, the occurrence of ocelli at the polyp stage is only known in the species of Stylocoronella (Scyphozoa, Stauromedusae). The light-sensitive organs of S. riedli are ultrastructurally investigated. In this interstitial-living species, each of the up to 24 ocelli is composed of between seven and nine monociliary sensory cells and between one and four pigment cells. A striking feature of the photoreceptive cilia is their peculiar axonemal pattern. This is expressed (a) by the presence of a third central microtubule at a certain point and (b) by the balloon-like swelling of the distal portion of the cilium, with clearly scattered microtubules in this area. Although the polyps of S. riedli show no distinct reaction to light stimuli, the ultrastructural results corroborate the hypothesis that these organs are light-sensitive organs. The possible function of the pigment granules is discussed.Abbreviations bb basal body - c cilium - co collar - csv crescent-shaped vesicle - cv clear vesicle - dcv dense-core vesicles - k kinetosome - m mitochondrion - mvb multivesicular body - n nucleus - oc ocellus - pc piment cell - pg pigment granule - sc sensory cell - sr striated rootlet - v vesicle     

Possible Ascidian Counterpart to the Vertebrate Saccus Vasculosus with Reference to Pyura tessellata (Forbes) and Boltenia echinata (L.)

The morphology of the hitherto unknown larva of Pyura tessellata (Forbes) is described and the discovery of an auxiliary brain vesicle is presented. The vesicle was also found in another pyurid species Boltenia echinata (L.). The auxiliary vesicle communicates with the sensory vesicle at the level of the statocyte and is lined with cubodial epithelial cells carrying a 2 μm globular structure projecting into the lumen. These cells very closely resemble primitive tunicate coronet cells previously reported and coronet cells typical for the saccus vasculosus found in elasmobranchs and many ganoids and teleosts.     

Regulation of Enzyme Synthesis of the Glyoxylate,the Citric Acid,and the Methylcitric Acid Cycles in Candida lipolytica

Syntheses of the key enzymes of the glyoxylate cycle, in Candida lipolytica, were highly repressed by glucose. Syntheses of the key enzymes of the methylcitric acid cycle were also slightly repressed by glucose but the degrees of repression in the syntheses of these enzymes were nearly equal to those of repression in the syntheses of several enzymes of the citric acid cycle. All enzyme syntheses repressed by glucose were derepressed during incubation with succinate as well as with n-alkanes: enzyme syntheses of the methylcitric acid cycle did not necessitate the addition of propionate or odd-carbon n-alkanes. The enzymes of the methylcitric acid cycle seem to be constitutive, similarly as those of the citric acid cycle.

In the parent strain, the respective enzyme levels of the cells grown on an odd-numbered n-alkane were similar to those of the cells grown on an even-numbered n-alkane. But in the mutant strain lacking 2-methylisocitrate lyase, the cells grown on the odd-numbered alkane contained aconitate hydratase, NADP-Iinked isocitrate dehydrogenase, isocitrate lyase, 2- methylcitrate synthase and 2-methylaconitate hydratase all at higher levels than the cells grown on the even-numbered alkane. Both the parent cells and the mutant cells grown on the same carbon source contained at individually similar levels of the following six enzymes; citrate synthase, NAD-linked isocitrate dehydrogenase, succinate dehydrogenase, fumarate hydratase, malate dehydrogenase, and malate synthase. The pleiotropic changes of enzyme activities in the mutant cells grown on the odd-numbered alkane seem to be ascribable to direct or indirect stimulation caused by threo-d_s-2-methylisocitric acid accumulation.     

Effects of soil treatments and cultivars on 'early dying' disease of potatoes caused by Globodera rostochiensis and Verticilliurn dahliae

Plots in a field infested with Globodera rostochiensis and Verticillium dahliae were treated in 1971 with methyl bromide, dazomet, aldicarb, benomyl or aldicarb + benomyl and potatoes (cv. Pentland Dell) were grown in four consecutive years. In 1971, all treatments delayed the development of haulm symptoms, decreased soil populations of G. rostochiensis and increased yields. In the second year yields were increased by all treatments except dazomet whereas in the third and fourth years only benomyl or aldicarb + benomyl increased yields. Soil populations of V. dahliae were decreased by methyl bromide in 1971 but not by other treatments or in later years. In 1976, Pentland Dell and Pentland Crown were grown on an infested field in plots fumigated with methyl bromide. Fumigation slightly delayed shoot emergence but increased plant height, ground cover and the size and persistence of the leaves; development of haulm symptoms was delayed and yield increased. Fumigating with methyl bromide at five times the normal rate was more beneficial to Pentland Dell than to Pentland Crown. In a comparison of 10 cultivars in 1975, symptoms developed sooner and yields were smaller on a field infested with G. rostochiensis and V. dahliae than on a non-infested field and on both sites Maris Peer plants died soonest and yielded least. Pentland Crown yielded most on the non-infested field and Maris Piper on the infested field.     

Reconstitution of the Z-Disk

Methanol-utilizing bacteria, Klebsiella sp. No. 101 and Microcyclus eburneus could grow aerobically and statically on 1,2-propanediol. The authors examined the presence of enzyme activity of adenosyl-B₁₂ dependent diol dehydratase as well as NAD dependent diol dehydroagenase. Adenosyl-B₁₂ dependent diol dehydratase activity was not detected in these organisms, even if these grown statically.

The dehydrogenase activity was found in the extract from these methanol-utilizing bacteria cells grown on various carbon sources. The partially purified enzyme preparation from the cells of Mic. eburneus grown aerobically on 1,2-propanediol dehydrogenated 1,2-propanediol, 1,2-butanediol and 2,3-butanediol. The enzyme activity was separated into two fractions, namely enzyme I and II on DEAE-Sephadex A-25 column chromatography. The enzyme I was different from the enzyme II in the ratio of enzyme activity to 1,2-propanediol and 2,3-butanediol, heat stability, pH stability and pH optimum, and effect of 2-mercaptoethanol.     

Electron microscopic investigations of zymomonas mobilis cells grown in low and high glucose concentrations

Summary Zymomonas mobilis cells grown in the presence of low and high glucose concentrations were examined by electron microscopy. Using ultrathin sectioning and agar diffusion method, significant changes in morphology were observed. Although the fine structure resembles that of a typical gram-negative bacterium, changes in glucose concentration and phases of growth lead to large cell wall vesicle or blebs formation. The possible implications of this morphological change to glucose uptake and ethanol formation are discussed.