Comparison of various atmospheric conditions for isolation and subcultivation of Mycoplasma hyorhinis from cell cultures

The efficiency of aerobic incubation was compared with incubation under various oxygen and carbon dioxide conditions for the isolation and subcultivation of three strains of Mycoplasma hyorhinis from VERO-cell cultures and subcultivation of three laboratory strains. Under anaerobic conditions with a low oxidation-reduction potential (at or below -115 mV) as obtained in jars, with catalysts, containing mixtures of 5%-10% CO2 in H2, very poor or no growth of any of the six M. hyorhinis strains was observed. When traces of oxygen were present (that is, under conditions with higher oxidation-reduction potentials, e.g. when omitting the catalyst in the above gas mixtures or in 5% CO2 + 95% N2) isolation from cell cultures was successful in most tests, but subcultivation of these primary isolates was seldom possible under these semi-anaerobic conditions. However, in most cases these primary isolates could be subcultivated aerobically, although aerobic conditions were unsatisfactory for isolation in about half of the experiments. Isolation of M. hyorhinis was optimal in 5% O2 + 95% N2, under which condition the isolates could also always be subcultivated. Isolation failed occasionally when 5% O2 + 5% CO2 + 90% N2 was used, thus indicating that 5% CO2 was slightly inhibitory. 5% CO2 in air and 10% CO2 either in air, H2 or N2 were also inadequate for isolation from cell cultures. In contrast to the findings with these cell culture-adapted M. hyorhinis strains, the laboratory strains could be subcultivated easily under all conditions tested except those with an oxidation-reduction potential at or below -115 mV; 100% CO2 was inhibitory for all 6 strains. Our findings may partly explain why M. hyorhinis is often considered "non-cultivable" on artificial media once adapted to cell cultures. The findings emphasize the need to employ also a micro-aerophilic condition (5% O2 in 95% N2) in the examination of cell cultures for mycoplasma.     

Different structure of polytene chromosomes ofPhaseolus coccineus suspensors during early embryogenesis

Summary The pattern of DNA and RNA puffs in pair VII of polytene chromosomes has been investigated in the suspensor ofPhaseolus coccineus during early embryo development. The pattern of³H-TdR and³H-U incorporation has been also detected. Collected data indicate that: 1. both heterochromatic regions, p₁₁ and q_(111+112), of chromosome pair VII, organize large DNA puffs; 2. DNA puffs of both regions are specific of different embryo differentiation steps; 3. a seasonal influence on the DNA puffing seems also to be present, as demonstrated by the comparison of the results collected in two different crops; 4. the incorporation experiment by³H-TdR evidences that not all DNA puffs show clustered labeling; 5. the RNA puffing of the two regions seems also to be specific of determined embryo stages.     

Action of two juvenile hormone antagonists on lepidopteran epidermis

The juvenile hormone antagonist ETB (ethyl-4-2(t-butylcarbonyloxy)-butoxybenzoate) caused formation of precocious larval-pupal intermediates after the 4th (penultimate)-larval instar of the tobacco hornworm, Manduca sexta, when 50 μg were applied to any 3rd stage larvae or to 4th stage larvae within 12 hr after ecdysis. This dose was most effective within 12 hr after ecdysis to the 3rd stage. In the black mutant larval assay for juvenile hormone, ETB had activity, 0.75 μg per larva giving half-maximal score. In vitro ETB acted as a juvenile hormone to prevent the ecdysteroid-induced change in commitment at concentrations above 0.1 μg/ml with an ED₅₀ at 2.8 μg/ml and as a partial juvenile hormone antagonist to 0.1 μg/ml juvenile hormone I at concentrations between 10^?3 and 10^?2 μg/ml. By contrast, EMD (ethyl-E-3-methyl-2-dodecenoate) had little juvenile hormone-like activity in vitro up to its limits of solubility (100 μg/ml) and exhibited sporadic partial juvenile hormone antagonistic activity in vitro at concentrations between 1 and 100 μg/ml. Since these concentrations were 10–1000 times that of juvenile hormone I in the medium, EMD apparently is not an efficient competitor.     

Translation of poly(U) on ribosomes from Streptomyces aureofaciens

Slowly cooled cells of Streptomyces aureofaciens contained mainly tight-couple ribosomes. Maximum rate of polyphenylalanine synthesis on ribosomes of S. aureofaciens was observed at 40°C, while cultures grew optimally at 28°C. Ribosomes of S. aureofaciens differed from those of E. coli in the amount of poly(U) required for maximum synthetic activity. The polyphenylalanine-synthesizing activity of E. coli ribosomes was about 3-times higher than that of S. aureofaciens ribosomes. The addition of protein S1 of E. coli or the homologous protein from S. aureofaciens had no stimulatory effect on the translation of poly(U). In order to localize alteration(s) of S. aureofaciens ribosomes in the elongation step of polypeptide synthesis we developed an in vitro system derived from purified elongation factors and ribosomal subunits. The enzymatic binding of Phe-tRNA to ribosomes of S. aureofaciens was significantly lower than the binding to ribosomes of E. coli. This alteration was mainly connected with the function of S. aureofaciens 50 S subunits. These subunits were not deficient in their ability to associate with 30 S subunits or with protein SL5 which is homologous to L7/L12 of E. coli.     

Studies on the subunit structure and amino acid sequence of triose phosphate isomerase from chicken breast muscle

1. Triose phosphate isomerase was prepared by chromatography on DEAE-cellulose of an (NH(4))(2)SO(4) fraction of an extract of homogenized chicken breast muscle. The product is homogeneous on gel electrophoresis and is suitable for growing crystals for X-ray work. The specific activity is 10000 units/mg and the value for E(0.1%) (280) is 1.20. 2. Comparison between the sum of the amino acid compositions of the tryptic peptides of the protein and the amino acid composition obtained on total hydrolysis of the protein indicates that the relative subunit mass is about 27000. 3. These data, together with the results of the examination of the amino acid compositions of a number of minor peptides, the number of peptides in the tryptic digest and the complete amino acid sequences of the tryptic peptides (the determination of which is described here), give no indication that the subunits are dissimilar. 4. A tentative amino acid sequence is presented for the protein, in which the ordering of the tryptic peptides is derived by homology with the sequence of the rabbit muscle enzyme (Corran & Waley, 1973). 5. An appendix describes the use that was made of mass spectrometry in the determination of some of the sequences. Mass-spectrometric data have been obtained for 35 residues, that is about 15% of the total sequence of the protein. 6. An extended version of the present paper has been deposited as Supplementary Publication SUP 50025 at the British Library, Lending Division (formerly the National Lending Library for Science and Technology), Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1973) 131, 5.     

The problem of carotenoid biosynthesis in the taxonomy of genera Rhodotorula and Rhodosporidium

This report presents results in the composition of major carotenoids of various coloured mutants of the genusRhodotorula and of mating types of the genusRhodosporidium. The separation of carotenoid intermediates was carried out by thin layer chromatography using Silufol 254 Kavalier and the determination of eluated spots by spectrophotometry. There was found no difference in carotenoid composition of both mating typesa and α of individual species of the genusRhodosporidium. The vegetative and sexual reproduction ofRhodotorula andRhodosporidium can be separated from the carotenogenesis using 10^?4 mol diphenylamine. It was concluded that lycopene could be the intermediate to mono- and dicyclic carotenoids; in the case of partial inhibition of the dehydrogenation step the direct cyclization of neurosporene to β-zeacarotene can be expected. An unknown compound, probably lycopersene was found and was considered to be the precursor of phytoene. Phytoene and phytofluene were proved in all studied samples. Nutritional conditions (vitamins, sulfur amino acids, etc.) are able to shift the ratios between major carotenoids. Rhodotorula aurantiaca strains were observed to be auxotrophic mutants of various characters and the existence of this species as independent one, was denied.     

Bemerkungen zur Taxonomie vonVicia sepium L.

In der ArtVicia sepium L. kann man in Mitteleuropa und in südlichen Teilen von Nordeuropa drei Varietäten unterscheiden: 1. var.sepium mit breiten Blättchen und einem kahlen bis angedrückt behaarten Kelch, 2. var.montana Koch mit schmalen Blättchen und einem kahlen oder zerstreut angedrückt behaarten Kelch, und 3. var.eriocalyx ?elak. mit breiten bis mittelbreiten Blättchen und einem abstehend langbehaarten Kelch, noch mit einer niedrigen und einer hohen Wuchsform. Bei allen Proben wurde die Chromosomezahl 2n=14 festgestellt.     

Bemerkungen zur Taxonomie und Nomenklatur vonLotus krylovii Schischk. etSerg. undL. corniculatus L. subsp.frondosus Freyn

Lotus krylovii Schischk. etSerg. undL. corniculatus L. subsp.frondosus Freyn sind zwei unterschiedliche Taxa, gekennzeichnet durch einige morphologische Merkmale.L. krylovii ist am nächsten mitL. tenuis Waldst. etKit. verwandt;L. corniculatus subsp.frondosus gehört in den Umkreis der Unterart vonL. corniculatus L., die im östlichen Teil des Areals der Art verbreitet ist. Nach ihrer Haupt-Verbreitung gehören die beiden Taxa zu den westasiatischen Arten.     

The genusSaccharomyces (Meyen) Reess

The authors submit a taxonomic evaluation of an intermediate group of strains between the speciesSaccharomyces carlsbergensis Hansen andSaccharomyces cerevisiae Hansen. The material consisted of atypical strains of “bottom” brewer’s yeasts and the synonymous strainsSaccharomyces monacensis Hansen andSaccharomyces mandshuricus Saito. It was found that there were two different serological types in the speciesSaccharomyces carlsbergensis, one of which was characterized by the presence of antigen “C” and was typical for this species, while the other possessed antigen “M” and was grouped roundSaccharomyces monacensis. This second serological type merges with a group of strains which gives only one third fermentation of raffinose, so that it is actually an intermediate betweenSaccharomyces cerevisiae Hansen andSaccharomyces carlsbergensis Hansen and indicates the course of progressive development from the former species to the latter. No close similarity was found betweenSaccharomyces mandshuricus Saito and some of the strains of the transitional group or typical representatives of the two main species, and the authors therefore consider that there is some obscurity as to its synonymity withSaccharomyces carlsbergensis.