- Volume 136, Issue 8, 1990

Addition of glycine betaine (1 mm) stimulated aerobic growth of Azospirillum brasilense Sp 7 in the presence of 0·3 m-NaCl. The nitrogenase activity of whole cells was particularly sensitive to salt stress, being almost totally inhibited in the presence of the same concentration of salt. Added glycine betaine strongly enhanced nitrogen fixation activity under salt stress. Under such conditions, maximal nitrogenase activity was obtained at a pO2 value (1 kPa) that inhibits nitrogen fixation activity in the absence of salt. We demonstrated the presence of a highaffinity transport system for glycine betaine, with an apparent K m of 10 μm. The osmolarity of the medium regulated the activity of the transport system. The maximal transport rates were 4 and 20 nmol min−1 (mg protein)−1 in cells grown in low-salt and high-salt medium, respectively. A high intracellular concentration of glycine betaine (480 μm) was observed only at a high osmolarity (0·3 m-NaCl). Glycine betaine uptake was significantly reduced in osmotically shocked cells and a glycine betaine binding activity was detected in the crude periplasmic shock fluid. This suggests a transport mechanism involving a periplasmic glycine betaine binding protein. A. brasilense was unable to use the transported glycine betaine as a carbon- or nitrogen-source, in low- or high-salt medium. Intracellular glycine betaine was not catabolized.

The production of exoenzymes which degrade cellulose, polygalacturonic acid and xylan was studied in mycelial and wall-less phenotypic derivatives of Neurospora crassa obtained by vegetative selection applied to a single fz; sg; os-1 (‘slime’-like) segregant (strain RCP-3) of a cross ‘slime’ × wild type. The unrelated stable ‘slime’ strain FGSC 1118 was also studied. The synthesis of polysaccharide-degrading enzymes was normally induced by polysaccharidic substrates and was sensitive to carbon-catabolite repression for both mycelium-forming phenotypes (mycelial intermediate and spheroplast-hyphal intermediate) of strain RCP-3. The stable ‘slime’ from RCP-3 produced cellulose-degrading activity and xylan-degrading activity constitutively but was fully sensitive to glucose repression. The stable ‘slime’ RCP-3 did not synthesize polygalacturonic-acid-degrading activity, even in the presence of inducers. For the stable ‘slime’ FGSC 1118, all of the polysaccharide-degrading activities were produced constitutively and were markedly resistant to glucose repression. The possible epigenetic origin of the different properties of stable ‘slimes’ RCP-3 and FGSC 1118 is considered. These results may relate to the role of the cell surface in the processing of regulatory signals which control the adaptation of the fungal cell to the nutritional environment.

Studies on the yeasts Candida oleophila, Candida utilis, Lipomyces starkeyi, Rhodosporidium toruloides and Saccharomyces cerevisiae revealed the existence of three different temperature adaptation responses involving changes in fatty acid composition. These conclusions were drawn by determining the growth rates, total cellular fatty acid content, fatty acid composition, degree of unsaturation, and the mean chain length of fatty acids over a range of growth temperatures. Within temperatures permitting growth, there were no changes in the major fatty acids of any of the yeasts, but the absolute amounts and relative compositions of the fatty acids did alter. In S. cerevisiae there were temperature-induced changes in the mean fatty acid chain length, whereas in R. toruloides there were changes in the degree of unsaturation. C. oleophila, C. utilis and L. starkeyi showed both responses, depending on whether the growth temperature was above or below 20–26 °C. Below 20–26 °C temperature-dependent changes were observed in the mean chain length whereas above 20–26 °C there were changes in the degree of unsaturation.

The uptake of pyrimidines and their derivatives into Candida glabrata and Candida albicans was measured using a novel technique in which the cells were rapidly separated from their suspending medium by centrifugation through a layer of an inert oil. The uptake of [14C]cytosine was linear for 30 s for all concentrations of pyrimidine tested. In C. glabrata but not C. albicans cytosine transport was mediated by both a high affinity (K M 0·8 ± 0·1 μm), low capacity [V 40 ± 4 pmol(μl cell water)−1 s−1] and a low affinity [K m 240 ± 35 μm], high capacity system [V770 ± 170 pmol (μl cell water)−1 s−1. The cytosine permease in C. glabrata was specific for cytosine and 5-fluorocytosine. In C. albicans there was only one cytosine transport system [K m 2·4 ± 0·3 μm; V50 ± 4 pmol (μl cell water)−1 s−1]; this system also transported adenine, guanine and hypoxanthine. Differences in nucleoside transport were also observed for C. glabrata and C. albicans, with the uridine permease in C. glabrata transporting only uridine and 5-fluorouridine whereas cytidine and adenosine were also transported by the uridine permease in C. albicans. Studies on the effect of nucleoside analogues on uridine transport in C. glabrata demonstrated the importance of the sugar moiety in determining the specificity of transport, with a hydroxyl residue on C-2 being apparently essential for transport.

Ribosomal DNA (rDNA) polymorphism was compared with electrophoretic enzyme polymorphism for the intra-and interspecies differentiation of Yersinia enterocolitica, Y. pseudotuberculosis, Y. intermedia, Y. aldovae, Y. frederiksenii and Y. kristensenii. DNA from 90 strains previously classified into six zymotypes (Y. enterocolitica and Y. frederiksenii) and into distinct enzyme electrophoretic patterns (the four other species) was digested with EcoRI or HindIII and analysed by Southern blotting. The six species were clearly differentiated from each other. In Y. enterocolitica, the subclassification of biotype 1 into zymotypes 1A and 1B was also reflected in the rDNA and the four other bio-zymotypes gave four different classes of restriction pattern. In Y. frederiksenii, both EcoRI and HindIII gave five distinct riboclasses which correlated with the zymotypes. In the four other species, the phenotype polymorphism appeared to be better correlated with the restriction fragment length polymorphism data in some enzymes than others. The data demonstrate that the inter- and intraspecies classification by rDNA polymorphism using two restriction enzymes is similar to that based on electrophoretic enzyme polymorphism. The analysis could be refined for taxonomic and epidemiological purposes by using other restriction enzymes.