Comparative studies on the regulation of DAHP synthetase activity in blue-green and green algae

Summary Regulation of DAHP synthetase activity was investigated in autotrophically grown blue-green and green algae. Members of the class of blue-green algae possess an enzyme, the activity of which is regulated by l-tyrosine and l-phenylalanine, whereby l-tyrosine is effective in 100 fold lower concentrations. DAHP synthetases of two organisms, Anabaena and Anacystis, were shown to belong to the V-type of allosteric enzymes.In contrast to the DAHP synthetase of blue-green algae regulation of this enzyme could not be demonstrated in two green algae, Ankistrodesmus and Maesotaenium. However, Euglena gracilis, both under conditions of mixotrophic and autotrophic growth, exhibits very effective regulation of this key enzyme; again, the inhibitors are tyrosine and phenylalanine. DAHP synthetase activity of Euglena has been purified about 40 fold; during this enrichment no separation of the enzyme activity inhibited by tyrosine and that by phenylalanine could be observed.     

Absorption and Screening in Phycomyces

In vivo absorption measurements were made through the photosensitive zones of Phycomyces sporangiophores and absorption spectra are presented for various growth media and for wavelengths between 400 and 580 mµ. As in mycelia, β-carotene was the major pigment ordinarily found. The addition of diphenylamine to the growth media caused a decrease in β-carotene and an increase in certain other carotenoids. Growth in the dark substantially reduced the amount of β-carotene in the photosensitive zone; however, growth on a lactate medium failed to suppress β-carotene in the growing zone although the mycelia appeared almost colorless. Also when diphenylamine was added to the medium the absorption in the growing zone at 460 mµ was not diminished although the colored carotenoids in the bulk of the sporangiophore were drastically reduced. Absorption which is characteristic of the action spectra was not found. Sporangiophores immersed in fluids with a critical refractive index show neither positive nor negative tropism. Measurements were made of the critical refractive indices for light at 495 and 510 mµ. The critical indices differed only slightly. Assuming primary photoreceptors at the cell wall, the change in screening due to absorption appears too large to be counterbalanced solely by a simple effect of the focusing change. The possibility is therefore advanced that the receptors are internal to most of the cytoplasm; i.e., near the vacuole.     

The histochemical demonstration of five lysosomal enzymes in the pars distalis of the amphibian pituitary

Summary The distribution of five lysosomal enzymes (acid phosphatase, -glucosaminidase, -glucuronidase, sulphatase and E 600-resistant esterases) was studied in the pars distalis of the urodelian pituitary. Naphthol-AS-compounds coupled with hexazonium-pararosanilin yielded particularly good localization. The typical lysosomal picture and a basically similar distribution were obtained throughout. The reaction product was mainly found in the globules of the globular basophils (-cells) reported as gonatropin producing cells by several authors. Some speculations on the possible function of the high amount of the lysosomal enzymes in the globular basophils are made.Dedicated to Professor W. Bargmann on his 60th birthday.This work was partly supported by a grant from Deutscher Akademischer Austauschdienst, Bad Godesberg.     

Ueber den stand der Agrarmeteorologischen Forschung in der Deutschen Demokratischen Republik

Ohne Zusammenfassung     

Spontane Mutationen bei Essigsäurebakterien

Zusammenfassung Mit einer modifizierten Penicillintechnik konnten aus Acetobacter-Populationen, die von einem prototrophen Klon stammten, spontane auxotrophe Mutanten isoliert werden. Bei sechs von acht untersuchten Stämmen wurden auxotrophe Mutanten gefunden. Die Häufigkeit dieser Mutanten betrug 10^-6.Die Auxotrophie, welche je nach Stamm auf Äthanol- oder Glucose-Medium auftrat, äußerte sich als Bedarf für Hefeextrakt. Hefeextrakt konnte durch Alanin, Prolin oder Glutaminsäure nicht ersetzt werden. Die Mutation von prototroph auf Äthanol-Medium zu auxotroph auf Äthanol-Medium hatte keine Änderung der spezifischen Aktivitäten der Schlüsselenzyme des Glyoxylsäurecyclus zur Folge.Es wurden auch spontane Mutanten gefunden, welche die partikelgebundene Äthanoldehydrogenase verloren hatten und dadurch unfähig waren, Äthanol zu oxydieren oder als C-Quelle zu verwerten.Bei den spontanen, auf Äthanol-Medium auxotrophen Mutanten handelt es sich um Mutationen von einer Art des Frateurschen Systems in eine andere. Mutanten, welche die partikelgebundene Äthanoldehydrogenase verloren haben, gehören definitionsgemäß nicht mehr in die Gattung Acetobacter. Die Bedeutung solcher interspezifischer und intergenerischer Mutationen für die Arteinteilung innerhalb der Essigsäurebakterien wird diskutiert.

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Spontaneous mutations in acetic acid bacteria

Summary 8 prototrophic Acetobacter-strains were examined for spontaneous auxotrophic mutants. Using a modification of the Penicillin technique, there were detected auxotrophic mutants in 6 strains. The mutant frequency was 10^-6.Auxotrophy manifested itself as a requirement for yeast extract. Alanine, proline and glutamic acid could not replace yeastextract. In addition to the auxotrophic mutants there were found spontaneous mutants, which had lost the particulate ethanol-dehydrogenase and therefore had become unable to oxidise ethanol.The mutation from prototrophic on minerals-ethanol medium to auxotropic on minerals-ethanol medium represents a mutation from one species of Frateur's system into an other species. Mutants which have lost the particulate ethanol-dehydrogenase belong no more to the genus Acetobacter. The importance of such interspecific and intergeneric mutations for the taxonomy of acetic acid bacteria is discussed.

     

Isolation of a Mutant of Escherichia coli with a Temperature-sensitive Fructose-1,6-Diphosphate Aldolase Activity

B?ck, August (Purdue University, Lafayette, Ind.), and Frederick C. Neidhardt. Isolation of a mutant of Escherichia coli with a temperature-sensitive fructose-1,6-diphosphate aldolase activity. J. Bacteriol. 92:464-469. 1966.-A mutant of Escherichia coli was isolated which was able to grow in rich medium at 30 C but not at 40 C. Upon exposure to 40 C, the cells immediately stopped ribonucleic acid (RNA) and deoxyribonucleic acid synthesis, but protein synthesis continued at a diminished rate for a short time. Addition of chloramphenicol did not release RNA synthesis from inhibition at 40 C. Synthesis of beta-galactosidase could be induced at high temperature despite the presence of glucose in the medium, indicating a lesion in glucose catabolism. Of many catabolic enzymes tested in cell-free extracts, only fructose-1,6-diphosphate aldolase activity appeared to be altered in the mutant cells. No activity was demonstrable in extracts of mutant cells grown at either 30 or 40 C, but determination of glucose-oxidation patterns revealed that the enzyme is probably active in vivo at 30 C. Temperature-resistant secondary mutants were found to have partially or fully restored aldolase activity, and temperature-resistant recombinants had normal aldolase activity, indicating that the growth pattern and the altered aldolase had a common genetic basis. Linkage data permitted the assignment of an approximate map location for the mutated aldolase gene.     

Elektronenmikroskopische Untersuchungen über Glaskörperrindenzellen und Zonulafasern

Zusammenfassung Es werden folgende Befunde zur Feinstruktur des Glaskörpers beim 16 Tage alten Rattenembryo und der Glaskörperrinde beiderseits der Ora serrata beim 3 Jahre alten, an Retinoblastom erkrankten Kind mitgeteilt:Der Glaskörper des Rattenembryos und die Glaskörperrinde des kindlichen Auges enthalten Fibroblasten. Sie unterscheiden sich nicht von den im Bindegewebe vorkommenden Fibroblasten. Im embryonalen Rattenglaskörper wurden außerdem faserbildende Zellen mit wabiger Struktur des Zytoplasmas beobachtet.Die Fibroblasten der Glaskörperrinde bilden die Fibrillen des Glaskörpergerüstes und der Zonulafasern. Diese Fibrillen zeigen eine deutliche Querstreifung. Die Streifung ist unregelmäßig oder periodisch. Die Länge der Perioden beträgt in unseren Schnitten bei den Glaskörperfibrillen des Rattenembryos und des kindlichen Auges meist etwa 120, seltener 210 A. An den Fibrillenbündeln einer Zonulafaser des kindlichen Auges wurden Perioden von 90–120, 400, 440 und 630 A beobachtet.Die Fibroblasten der Glaskörperrinde des kindlichen Auges liegen im Bereich der Pars plana corporis ciliaris auch tief in den Buchten und Falten des Ciliarepithels. Hierdurch wird eine maximal große Anheftungsfläche für die von ihnen produzierten Fibrillen der Zonulafasern gewährleistet.Die Pars plana corporis ciliaris des kindlichen Auges ist von einem dichten Netz von Fibroblastenfortsätzen überzogen. Auch vereinzelte Makrophagen finden sich hier.Unsere elektronenmikroskopischen Befunde bestätigen die Angaben von Balazs über das Vorkommen von Fibrocyten (Fibroblasten) und Makrophagen in der Glaskörperrinde. Ferner bestätigen sie die bereits von früheren Autoren lichtmikroskopisch gewonnenen Erkenntnisse, wonach es sich beim Glaskörper um mesenchymales Gewebe, bei den Zonulafasern im Bindegewebsfasern handelt.

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Summary The following electron microscopical findings in the vitreous body of 16-day-old rat embryos and in the vitreoretinal border layer on both sides of the ora serrata in a 3-year-old child are reported:The vitreous body of the rat embryo and the vitreoretinal border layer of the infant eye contain fibroblasts. These fibroblasts do not differ from those present in connective tissue. The embryonic vitreous body of the rat contains fibre-forming cells, which show an alveolar structure of the cytoplasm.The fibroblasts in the cortical tissue layer of the vitreous body form the fibrils of the stroma of the vitreous body and the zonula fibres. These fibrils show a marked cross striation. The striation is either irregular or shows periodicity. In the vitreous body of the rat embryo and of the infant eye the lenght of these periods has been measured with 120 and 210 A. Periods of 90–120, 400, 440 and 630 A could be shown in the fibrillar bundles of a zonula fibre of the infant eye. In the region of the pars plana corporis ciliaris the fibroblasts of the cortical tissue layer of the vitreous body of the infant eye are also found deep down in the sinus and folds of the ciliary epithelium. Thus they guarantee an as large as possible area for the attachment of the fibrils of the zonula fibres which they produce.The pars plana corporis ciliaris of the infant eye is covered with a dense network of fibroblast processes. Also cells of the macrophage type may be found in this region.Our electron microscopical findings confirm those of earlier light microscopists and lately by Balazs. The vitreous body of the eye is formed by mesenchymal tissue, whereas the zonula fibres are formed by connective tissue.

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Mit dankenswerter Unterstützung durch den Schweizerischen Nationalfonds zur Förderung der Wissenschaftlichen Forschung.     

Production of doubled haploid lines in frequencies sufficient for barley breeding programs

Winter barley (Hordeum vulgare L.) anthers were cultured on different liquid and on starch-solidified media. The optimal embryo and callus formation with different F₁-lines and the cv. Igri was obtained on a liquid medium with 20% Ficoll, 20 g/l maltose and barley starch. But the influence of the growth conditions of the donor plants and the genotypical differences are still enormous. The procedure has been optimized to such an extent that it can be used economically.