Action of Bacillus subtilis alpha-amylase on native wheat starch

Native starch granules from wheat have been subjected to enzymatic depolymerization with an alpha-amylase from Bacillus subtilis. Crystallites made from short-chain amylose and residues from mild acid hydrolysis have been also tested. Electron microscopy, particle size analysis, DSC, and x-ray diffractometry reveal that enzymatic degradation occurs granule by granule. Gel permeation chromatography shows off the macromolecular nature of the remaining material. In contrast, acid erodes simultaneously all the granules, leading to a splitting into small particles. Crystalline fractions are completely degraded by alpha-amylase. These results support evidence for an active disentanglement of chains, carried out by the different subsites of alpha-amylase molecules. A simple mathematical treatment is proposed to explain the results of the kinetics.     

Pigment patterns in mutants affecting the biosynthesis of pteridines and xanthommatin in Drosophila melanogaster

Eye-color mutants of Drosophila melanogaster have been analyzed for their pigment content and related metabolites. Xanthommatin and dihydroxanthommatin (pigments causing brown eye color) were measured after selective extraction in acidified butanol. Pteridines (pigments causing red eye color) were quantitated after separation of 28 spots by thin-layer chromatography, most of which are pteridines and a few of which are fluorescent metabolites from the xanthommatin pathway. Pigment patterns have been studied in 45 loci. The pteridine pathway ramifies into two double branches giving rise to isoxanthopterin, drosopterins, and biopterin as final products. The regulatory relationship among the branches and the metabolic blockage of the mutants are discussed. The Hn locus is proposed to regulate pteridine synthesis in a step between pyruvoyltetrahydropterin and dihydropterin. The results also indicate that the synthesis and accumulation of xanthommatin in the eyes might be related to the synthesis of pteridines.Support for this work was provided to J.F. in part by a grant from the Ministerio de Universidades e Investigación (Spain) and to F.J.S. by a grant from the Ministerio de Educación y Ciencia (Spain).     

Membrane Operational Impedance Spectra in Chara corallina Estimated by Laplace Transforms Analysis

The membrane operational impedance spectrum of Chara corallina Klein ex Willd. (R. Brown) cells is investigated using Laplace transform analysis. The spectrum changes with both amplitude and sign of the electrical stimulation when time- and voltage-dependent K⁺ channels contribute to the membrane conductance. We compare the advantages and disadvantage of this technique for studying membrane impedance with those of the alternating current method and the white noise method.     

Yeast species utilizing uric acid,adenine,n-alkylamines or diamines as sole source of carbon and energy

Yeast strains utilizing uric acid, adenine, monoamines or diamines as sole source of carbon and energy were isolated from several soil samples by the enrichment culture method. The most common species wasTrichosporon cutaneum. Strains ofCandida catenulata, C. famata, C. parapsilosis, C. rugosa, Cryptococcus laurentii, Stephanoascus ciferrii andTr. adeninovorans were also isolated. All strains utilizing uric acid as sole carbon source utilized some primaryn-alkyl-l-amines hydroxyamines or diamines as well. The ascomycetous yeast strains showing these characteristics all belonged to species known to assimilate hydrocarbons. Type strains of hydrocarbon-positive yeast species which were not found in the enrichment cultures generally assimilated putrescine, some type strains also butylamine or pentylamine, but none assimilated uric acid. Methanol-positive species were not isolated. Type strains of methanol-positive and of hydrocarbon-negative species did not assimilate uric acid, butylamine or putrescine. Assimilation of putrescine as sole source of carbon and energy may be a valuable diagnostic criterion in yeast taxonomy.     

Microhabitat utilization by two leptodactylid frogs in the Andes of central Chile

Summary Some basic parameters of the life history of Alsodes montanus and Alsodes tumultuosus (Anura-Leptodactylidae), were studied from 1977 to 1980 by periodic field observations at Farellones and La Parva (33–34° south lat.; 2,700–3,000 m above sea level). Special attention was paid to strategies of resource partitioning in relation to gross features of the environment. The latter was unstable with a relative short period favorable for activity of the animals. Physical environmental differences between the first and second season of this study, resulted in a decrease in total number of active adults, a reduction in the duration of larval activity and a shift in microhabitat preferences of larvae.During the favorable season, October to May, adults of both species showed spatial and temporal segregation, related to different physical features of the environment; larvae did not show temporal segregation. Larvae of both species were found in seven different microhabitats; only in one of these did they show significant difference in microhabitat preference, A. tumultuosus was found more often in crevices. Microhabitat dimensions were more important than time and food resources in the separation of the niches of the two species. The segregation of niche dimensions, microhabitat, diel and annual activity and food were not complementary.Coexistence was therefore observed with the species tending to use different resources. When the same resource was used, it was not limiting.     

Isolation and cryopreservation of O-methylthreonine-resistant Rosa cell lines altered in the feedback sensitivity of l-threonine deaminase

O-Methylthreonine (OMT) inhibits the growth of plated Rosa cells (ID₅₀6·10^-6M). Isoleucine is able to reverse efficiently and specifically this OMT toxicity. From OMT-resistant colonies occurring at a frequency of 1.58·10^-7 variants per cell plated at 10^-4M OMT, the variant strains OMT^R-1 and OMT^R-2 were isolated, cloned via protoplasts and characterized. Both variants were ten times more resistant to OMT than the wildtype and were cross-resistant to another isoleucine analog, dl-4-thiaisoleucine. The resistant variants retained their resistance after storage for three years in liquid nitrogen. Both resistant strains were stable for several months when subcultured in the absence of OMT although it was shown in a reconstitution experiment that wildtype cells overgrow OMT^R-2 variant cells if co-cultivated for many passages in drug-free medium. One case of instability was observed upon long-term subculturing in drug-free medium: the strain OMT^R-1D^* partially lost phenotypic properties. Resistance to OMT was followed qualitatively by a new method based on inhibition-zone formation in cell suspensions plated in agar medium. The OMT-resistant variants showed a reduction in sensitivity of the enzyme l-threonine deaminase to feedback inhibition by isoleucine, a decreased stability of l-threonine deaminase when stored at-18°C or incubated at +55°C and a two- to threefold increase of the free isoleucine pool within the cells. The genetical events and the biochemical mechanisms which might lead to the observed stable and biochemically defined character are discussed with particular reference to the high ploidy level of the Rosa cell line.Abbreviations OMT l-O-methylthreonine - TD l-threonine deaminase     

Vegetation succession on the dunes near Oostvoorne,The Netherlands,since 1934, interpreted from air photographs and vegetation maps

The vegetation succession on the dunes near Oostvoorne, The Netherlands has been followed by means of a novel combination of repeated large-scale vegetation mapping and air photograph interpretation. Vegetation units have been discerned on the formation level because these could be distinguished fairly easily on the photographs and because the rates of change are appropriate to the time interval chosen. Nineteen formations were distinguished. Five 1:6250 maps were constructed, reflecting the formation pattern in 1934, 1943, 1959, 1972 and 1980. An overlay with 2736 grid points at 25 m field distance was used to quantify changes in the formation pattern.The results suggest a pronounced multiple pathway succession with nevertheless three principal trajectories of succession from pioneer to woodland vegetation. The outer dunes, which have originated since 1910, are distinct in successional characteristics from the inner dunes, which already existed but were released from heavy grazing pressure in 1910. The rate of change in the outer dunes was high in the beginning and is slowly decreasing eversince. In the inner dunes it went the other way around. Through visual extrapolation the likely formation patterns in 1910 and in 2000 were estimated.Transition frequencies proved highly variable for most formations. Moreover, strong spatial dependence was found. Limitations in the use of Markov models in cases of long-term succession in heterogeneous environments are discussed.Nomenclature follows the same sources as in van der Maarel et al. (1984).Field work carried out 1980–1981 when the authors were at the Division of Geobotany, University of Nijmegen. We thank Jos Rijntjes, Nijmegen, for his cooperation in the field. Vegetation maps were prepared and calculations performed at Uppsala. We thank the Foundation Het Zuid-Hollands Landschap, Rotterdam, for providing facilities and a grant for fieldwork as well as additional means to reproduce the vegetation maps. Two reviewers gave useful comments.     

An improved method to isolate lichen algae by gel filtration

Photobiont cells of the lichen Evernia prunastri have completely been separated from their fungal partner by filtration through a bed of Sepharose 2B. Both mannitol and ribitol have been quantified by gas-liquid chromatography in the different steps of the isolation procedure. Absence of mannitol, which is exclusively produced by the mycobiont, has been used as the best probe to monitor isolation.     

Immunoelectrophoretic approach to the metabolic regulation of glutamine synthetase in Rhodopseudomonas capsulata E1F1: role of glutamine

Anti-glutamine synthetase serum was raised in rabbits by injecting purified glutamine synthetase (GS) of the phototrophic bacterium Rhodopseudomonas capsulata E1F1. The antibodies were purified to monospecificity by immunoaffinity chromatography in GS-sepharose gel. These anti-GS antibodies were used to measure the antigen levels in crude extracts from bacteria, grown phototrophically with dinitrogen, nitrate, nitrite, ammonia, glutamate, glutamine or alanine as nitrogen sources. The amount of GS detected by rocket immunoelectrophoresis was proportional to Mn²⁺-dependent transferase activity measured in the crude extracts. Addition of GS inhibitor l-methionine-d,l-sulfoximine (MSX) to the actively growing cells promoted increased antigen levels, that were not found in the presence of glutamine or chloramphenicol. The ammonia-induced decrease in GS relative levels was reverted by MSX. GS levels remained constant when phototrophically growing cells were kept in the dark.Abbreviations GS glutamine synthetase - MOPS 2-(N-morpholine) propane sulfonate - MSX l-methionine-d,l-sulfoximine