Recessive and dominant genes controlling inducible sexual agglutinability in Saccharomyces cerevisiae

Summary We have characterized the two dominant genes, IND 1 and IND 2, responsible for inducible sexual agglutinability. The strains carrying these genes differ from the inducible strains carrying the recessive gene, saa 1 in the following points. The former strains produce agglutination substance at 22°, 28°, and 37° C only in response to sex pheromone of the opposite mating type, but the latter strains produce the substance constitutively without the pheromone at 22° C, only in response to the pheromone at 28° C, and do not produce the substance, even in the presence of the pheromone, at 37° C.We suggest that strains carrying one of the dominant, inducible genes are wild type and have a pheromone-controlled regulatory system of sexual agglutinability.     

The growth of Epidermophyton floccosum and E. stockdaleae at different temperatures

The ability of 17 strains of genus Epidermophyton (15 strains belonging to Epidermophyton floccosum, one to E. floccosum var. nigricans and one to E. stockdaleae) to grow at different temperatures (4 °C, 25 °C, 28 °C, 31 °C, 34 °C, 37 °C and 40 °C) was stated.The strains were inoculated on Sabouraud Dextrose Agar and regularly controled over a period of 14 days when the plates were incubated at 25 °C, 28 °C, 31 °C, 34 °C, 37 °C and 40 °C, and over a period of 70 days when the temperature was 4 °C. The optimal growth of E. floccosum was observed at 28 °C and 31 °C, and no signs of growth were recorded neither at 4 °C nor at 40 °C. The optimal development of E. stockdaleae was observed at 25 °C and 28 °C. This species grew from 4 °C to 31 °C.     

Life history,resting egg formation,and hatching may explain the temporal-geographical distribution ofArtemia strains in the Mediterranean basin

Life history traits are presented for the sexual species (Artemia tunisiana) and for parthenogenetic diploid and tetraploid strainsA. parthenogenetica reared at 15 °, 24 ° and 29.5 °C.In laboratory cultures, we present evidence that the seasonal appearance of the sexualArtemia tunisiana (the dominant winter-spring population), and of two parthenogenetic populations ofArtemia (the dominant spring-summer populations) in certain Spanish saltworks is controlled by temperature through its effect on reproductive and survival traits. Minimum and maximum reproductive output and survival for the sexual and parthenogenetic populations, respectively, occurred at a typical temperature (24 °C) of the late-spring season when the sexual population is replaced by parthenogenetic forms. Furthermore, the high production and hatchability of cysts from the sexual population at low temperatures (15 °C), and of the parthenogenetic populations at middle temperature (24 °C), indicate the role of dormancy as an adaptation regulating seasonal occurrence.     

Regulation of the production of the agglutination substances responsible for sexual agglutination in Saccharomyces cerevisiae: Changes associated with conjugation and temperature shift

Summary Isolated zygotes showed self-agglutination caused by the sex-specific glycoproteins, the agglutination substances responsible for sexual agglutination. The agglutination substances of both a and mating types were detected in the extracts obtained by the autoclave method from zygotes. Although the first diploid daughter cells from zygotes showed self-agglutinability, the self-agglutinability decreased gradually in the successive diploid daughter cells. The self-agglutination in diploid cells was also brought about by the complementary binding of the sex-specific agglutination substances of opposite mating types.The constitutive sexual agglutinability in a and cells was lost with concomitant loss of the agglutination substances in both cell wall and cytoplasmic fractions when cultured at a temperature higher than 35°C.The repression of the production of the agglutination substances was reversed by the opposite mating type pheromones even at the repressive temperature, 36°C, associated with the appearance of sexual agglutinability. The sex pheromones, a substance-I and substance-I, and the binding substance for substance-I were produced even at 36°C, repressive for the production of the agglutination substances.     

Effect of incubation temperature on zearalenone production by strains of Fusarium crookwellense

After 6 weeks incubation on rice 2 strains of Fusarium crookwellense produced more zearalenone (6060–5010 mg/kg dry wt of culture) at ambient temperature (16–29°C) in daylight than at ambient temperature (18–23 °C) in darkness or at controlled temperatures of 11 °C, 20 °C or 25 °C in darkness. Yields at 25 °C were low. Incubation at 11 °C during the second 3 weeks incubation increased yields only when preliminary incubation had been at 25 °C. After 6 weeks incubation at controlled temperatures in darkness, 4 strains produced most zearalenone at 20 °C (2460-21 360 mg/kg), 1 strain at 11 °C (6570 mg/kg). Yields at a temperature oscillating daily from 10–20 °C were less than at 15 °C. One of the 5 strains produced appreciable amounts of a-zearalenol (1645 mg/kg at 20°C) and 2 of nivalenol (340 and 499 mg/kg at 20 °C).     

The sexual agglutination substance is secreted through the yeast secretory pathway in Saccharomyces cerevisiae

Summary A Saccharomyces cerevisiae a strain carrying the secretory mutation sec1, sec7 or sec18 showed no sexual agglutination ability when treated with pheromone at the restrictive temperature 36° C, although the a agglutination substance had accumulated in the cytoplasm. These cells became sexually agglutinable, with a concomitant decrease in the agglutination substance in the cytoplasm, when the temperature was shifted from 36° C down to the permissive temperature 24° C after the addition of, cycloheximide. The a agglutination substance was barely detectable in sec53 cells (a) treated with pheromone at 36° C, indicating that the active a agglutination substance was formed after the export of its precursor into the endoplasmic reticulum. These results indicate that the a agglutination substance is exported through the yeast secretory pathway and that pheromone acts at the level of synthesis of the precursor molecule of the substance. An strain carrying sec1, sec7 or sec18 behaved just like an a strain carrying the sec gene in the induction of agglutination ability by the opposite mating type sex pheromone.     

Assessment of the tolerance of Lactococcus lactis cells at elevated temperatures

When Lactococcus lactis strains were exposed directly to the lethal temperature of 50 C for 30 ;min, 0.1–31% of the cells survived. However, when pre-exposed to 40 °C, prior to exposure at 50 °C, 4–61% of the cells survived. A plasmid carrying a unique heat shock gene from the thermophile Streptococcus thermophilus was cloned into L. ;lactis. When the transformed cells were cultivated at 30 °C the introduction of the plasmid had no obvious effect on the growth of L. ;lactis. However, when the temperature was abruptly shifted from 30 °C to 42 °C at mid-growth phase the growth decreased by 50%.     

Degradation of phenol and toxicity of phenolic compounds: a comparison of cold-tolerant<Emphasis Type="Italic"> Arthrobacter</Emphasis> sp. and mesophilic<Emphasis Type="Italic"> Pseudomonas putida</Emphasis>

Phenol degradation efficiency of cold-tolerant Arthrobacter sp. AG31 and mesophilic Pseudomonas putida DSM6414 was compared. The cold-tolerant strain was cultivated at 10°C, while the mesophile was grown at 25°C. Both strains degraded 200 mg and 400 mg phenol/l within 48–72 h of cultivation, but the cold-tolerant strain produced more biomass than the mesophile. Both strains oxidized catechol by the ortho type of ring fission. Catechol 1,2 dioxygenase (C1,2D) activity was found intra- and extracellularly in the absence and in the presence of phenol. In the presence of 200 mg phenol/l, C1,2D activity of the mesophile was about 1.5- to 2-fold higher than that of the cold-tolerant strain. However, an initial phenol concentration of 400 mg/l resulted in a comparable enzyme activity of the cold-tolerant and the mesophilic strain. The two strains differed significantly in their toxicity pattern towards 12 aromatic (mostly phenolic) compounds at different growth temperatures, which was determined via growth inhibition in the presence of nutrients and toxicants. For the cold-tolerant strain, toxicity was significantly lower at 10°C than at 25°C. The mesophile showed a significantly lower susceptibility to high hydrocarbon concentrations when grown at 25°C compared to 10°C.Communicated by K. Horikoshi     

Evaluation of host and pathogen responses for screening soil amendments to control Sclerotium rolfsii

Three strains of Bradyrhizobium, 280A, 2209A and 32H1, that nodulated peanuts (Arachis hypogaea L.), were tested for their ability to grow and survive at elevated temperatures of up to 42°C in laboratory culture. Strain 32H1 was unable to grow at 37°C and was more sensitive to elevated temperatures than the other two strains. All three produced heat-shock proteins of molecular weights 17 kDa and 18 kDa. Two greenhouse experiments were conducted to determine the effect of high root temperature on nodulation, growth and nitrogen fixation of peanut. Two peanut varieties (Virginia cv NC7 and Spanish cv Pronto) were inoculated and exposed to root temperatures of 30°, 37° and 40°C. Nodulation and nitrogen fixation were strongly affected by root temperature but there was no variety × temperature interaction. At a constant 40°C root temperature no nodules were formed. Nodules were formed when roots were exposed to this temperature with diurnal cycling but no nitrogen fixation occurred. Highest plant dry weight, shoot nitrogen content and total nitrogen were observed at a constant root temperature of 30°C. Increasing root temperature to 37°C reduced average nitrogen content by 37% and total nitrogen by 49% but did not reduce nodulation. The symbiotic performance of the strains corresponded to their abilities to grow and survive at high temperature in culture.     

Prolonged environmental stress via a two step process selects mutants of Escherichia,Salmonella and Pseudomonas that grow at 54°C

A prolonged incubation of Escherichia, Salmonella or Pseudomonas at 48°C with nalidixic acid selected mutants (T48) able to grow at 48°C. A prolonged incubation at 54°C of the T48 mutants selected mutants (T54) able to grow at 54°C. These mutants were susceptible to the same bacteriophages as the original mesophilic strains. Auxotrophic phenotypes of Escherichia coli and Salmonella typhimurium mesophilic parents were demonstrated by these mutants if they were cultivated on minimal agar with cellobiose at 48°C or 54°C or on a minimal agar with glucose at 37°C. The T48 alleles mapped in the gyrA region of E. coli or S. typhimurium chromosome. In S. typhimurium the T54 alleles, which permit growth at 54°C, were shown by cotransductional analysis to be linked to gyrA.     

Characterization of proteases produced by newly isolated and identified proteolytic microorganisms from fermented fish (Budu)

Eight different strains ofBacillus were isolated from fermented fish (Budu) and their proteolytic enzyme activities were determined after 18 h cultivation at room temperature (35° C). Four isolates possessed high protease activities. Optimum pH for these enzymes was between 7.0 and 8.0 and the optimal temperature was 55° C. The proteases retained 40% of their original activity after 20 min at 55° C but lost all activity at 65° C. Three of the four isolates were identified asBacillus subtilis, the fourth asBacillus licheniformis.     

Chemotactic behavior of Campylobacter spp. in function of different temperatures (37 °C and 42 °C)

The chemotactic behaviour of Campylobacter strains was determined in the presence of different amino acids at two temperatures (37 °C and 42 °C). Two strains of catalase positive (Campylobacter jejuni) and negative (Campylobacter sputurum) Campylobacter were isolated from river water in Tonekabon, Iran and identified by phenotyping and 16srRNA Gene sequencing methods. Chemotactic responses of the isolates were assessed toward a variety of amino acids viz., L-cystine, L-asparagine, L-histidine, L-aspartic acid, L-serine, L-phenylalanine, L-leucine and L-tryptophan by disc and capillary methods at two temperatures: 37 °C and 42 °C. C. jejuni showed positive chemotactic response towards L-cystine,L-tryptophan, L-phenylalanine, - L-leucine, L-asparagine and L-Serine at both, 37 °C and 42 °C however, it was greater at 37 °C. C. sputurum showed negative or weak response towards all of the amino acids. In addition, C. jejuni illustrated strong chemotactic response to L-asparagine follow by L-serine and weak chemotaxis response to L-phenylalanine and L-cysteine at 37 °C. Overall, C. jejuni showed relatively strong chemotactic response to some amino acids, likewise it was greater at 37 °C. Hence, the human body temperature (37 °C) in compared to avian body temperature (42 °C) probably promotes chemotactic response of C. jejuni, which it might be a reason for causing disease in human being compared to avian.     

Reduction of the nodulation barrier inMedicago laciniata by alteration of the root temperature

Medicago laciniata, an annual leguminous plant of Saharo-Sindian origin, is particularly refractory to root nodulation by most strains ofRhizobium meliloti. Using a series of such bacterial strains belonging to the 8 groups of Brockwell and Hely, and a variety of environmental conditions, it was noted that several normally non-nodulating strains (at 20°C) produced ineffective nodules at root temperatures of 24°C to 28°C. Nodulation at 20°C failed to occur in the presence of a wide variety of test compounds and physical conditions. No phytoalexins or anti-Rhizobium growth inhibitors were isolated from inoculated root tissue at any temperature. Temperature shift experiments indicated no infection of the root hairs at 20°C, and infection threads produced at the permissive root temperature failed to elongate after transfer to 20°C. However, if meristematic activity had been initiated in the inner root-cortical cells as a result of infection thread penetration at 28°C, no blockage of nodule maturation occurred upon subsequent transfer to 20°C root temperature. Nodules produced at 28°C were completely devoid of nitrogenase activity, although the apical (but not the distal) regions contained normal-appearing bacteriods, surrounded by enclosing membranes, and possessed a fully functional leghaemoglobin. A shortage of metabolic energy did not appear to be involved in the ineffective response. A hypothesis to explain the nodulation phenomenon observed was based on the observation in the roots of 2 factors present at 20°C but not at 28°C.     

SelectingRhizobium phaseoli strains for use with beans (Phaseolus vulgaris L.) in Kenya: Tolerance of high temperature and antibiotic resistance

Forty one strains ofRhizobium phaseoli were screened for the ability to multiply at high temperatures on yeast extract-mannitol agar. Most strains were tolerant of 30°C, eight strains were tolerant of 45°C and two of 47°C although the rate of multiplication was reduced at 45–47°C. The high temperature-tolerant strains were isolated from Kenyan soils and were fast-growing. Seven of the eight strains tolerant of 45–47°C lost their infectiveness after incubation at high temperature but four strains tolerant of 40°C remained infective after incubation at that temperature.Thirty six strains were resistant to 200 g ml^–1 streptomycin sulphate and 29 strains to 200 g ml^–1 spectinomycin dihydrochloride. Eight strains were resistant to both antibiotics each at 200 g ml^–1. Two of the double-labelled antibiotic-resistant mutants lost their infectiveness onPhaseolus vulgaris. The response to acidity was unaltered and two of the mutants showed a decrease in temperature tolerance. The doublelabelled mutants were recoverable from two Kenyan soils.     

Survival kinetics of Salmonella enterica serotype senftenberg (S. senftenberg) after heat and acid stress

The survival kinetics of two clinical isolates of Salmonella senftenberg were studied after heat and acid stress. The strains survived better at 53 and 55 °C after heat shock of 30 min at 50 °C or overnight heat adaptation at 45 °C. An increase in the decimal reduction time, D, of heat-shocked [10.2 min (53 °C) and 9.37 min (55 °C)] and heat-adapted [8.12 min (53 °C) and 7.8 min (55 °C)] cells was observed compared with the non-stressed cells [6.87 min (53 °C), 6.56 min 55 °C)]. A significant difference was also observed in the survival of acid-adapted to acid non-adapted S. senftenberg bacteria.     

Infectivity of a Bulgarian and an American strain ofSteinernema carpocapsae against codling moth

A strain of Steinernemacarpocapsae obtained from overwintering codling mothscollected in Bulgaria was identified to species usingthe polymerase chain reaction. The infectivity ofthis strain towards codling moth prepupae was comparedto that of a U.S. strain at three temperatures (11, 15and 20 °C). Infectivity was low at 11 °C with 8%mortality recorded for both strains. Mortality was61% and 62% at 15 °C, and 82% and 81% at 20 °C forthe Bulgarian and U.S. strain, respectively. The meanpercent mortality at each individual temperature wassignificantly different from each other whenmortalities of the two strains were combined. Thus,the Bulgarian strain did not provide a low temperatureinfectivity advantage compared to the U.S. strain.     

Modulation of virulence and antibiotic susceptibility of enteropathogenic Escherichia coli strains by Enterococcus faecium probiotic strain culture fractions

The increasing rate of antimicrobial resistance drastically reduced the efficiency of conventional antibiotics and led to the reconsideration of the interspecies interactions in influencing bacterial virulence and response to therapy. The aim of the study was the investigation of the influence of the soluble and cellular fractions of Enterococcus (E.) faecium CMGB16 probiotic culture on the virulence and antibiotic resistance markers expression in clinical enteropathogenic Escherichia (E.) coli strains.The 7 clinical enteropathogenic E. coli strains, one standard E. coli ATCC 25,922 and one Bacillus (B.) cereus strains were cultivated in nutrient broth, aerobically at 37 °C, for 24 h. The E. faecium CMGB16 probiotic strain was cultivated in anaerobic conditions, at 37 °C in MRS (Man Rogosa Sharpe) broth, and co-cultivated with two pathogenic strains (B. cereus and E. coli O28) culture fractions (supernatant, washed sediment and heat-inactivated culture) for 6 h, at 37 °C. After co-cultivation, the soluble and cellular fractions of the probiotic strain cultivated in the presence of two pathogenic strains were separated by centrifugation (6000 rpm, 10 min), heat-inactivated (15 min, 100 °C) and co-cultivated with the clinical enteropathogenic E. coli strains in McConkey broth, for 24 h, at 37 °C, in order to investigate the influence of the probiotic fractions on the adherence capacity and antibiotic susceptibility. All tested probiotic combinations influenced the adherence pattern of E. coli tested strains. The enteropathogenic E. coli strains susceptibility to aminoglycosides, beta-lactams and quinolones was increased by all probiotic combinations and decreased for amoxicillin-clavulanic acid. This study demonstrates that the plurifactorial anti-infective action of probiotics is also due to the modulation of virulence factors and antibiotic susceptibility expression in E. coli pathogenic strains.     

The release of sex-specific substances responsible for sexual agglutination from haploid cells of Saccharomyces cerevisiae

Cell surface substances responsible for sexual cell agglutination were successfully released in a large quantity from heterothallic haploid cells of Saccharomyces cerevisiae by a newly established autoclaving method. The conditions for this releasing phenomenon were examined. The sexual agglutination substances were solubilized most efficiently when the cells, suspended in a 30 mM Tris-HCl, pH 7.0, 5 mM EDTA solution, were autoclaved at a pressure of 1 kg/cm² at 120 °C for 3 min. The substances were specifically adsorbed by the cell surface of the opposite mating type, resulting in the masking of agglutinability of the cells of the opposite mating type. The substances were not released from the surface of cells which lacked sexual cell agglutination. The evidence suggesting the formation of a molecular complex between a- and α-agglutination substances in vitro is also presented. The above procedure is applicable to the solubilization of surfage agglutination substances from various strains of S. cerevisiae.     

Mutants inducible for sexual agglutinability in Saccharomyces cerevisiae

Summary We have isolated the mutants, T55s-41(a) and T562s-161 () which have no sexual agglutinability when cultured at 28°C, but become sexually agglutinable by the action of the sex pheromone produced by respective opposite mating type. The sex-specific glycoproteins responsible for sexual agglutination were detected in the mutants treated with the opposite mating type pheromone, but not in those treated with the same mating type pheromone.The induction of sexual agglutinability by the pheromone required both nitrogen and carbon sources and was inhibited by cycloheximide. The induction by the pheromone of sexual agglutinability was much more sensitive to osmotic shock and Triton X-100 in T55s-41 than in H1-0, an inducible a strain found in our stock cultures. When cultured at 22°C both T55s-41 and T562s-161 produced respective agglutination substances without the sex pheromones.H1-0 carried more than one genes responsible for the inducibility (inducible genes). The inducible genes carried by T55s-41 and T562s-161 were recessive, possibly linked to none of the mating type locus, thr4 and his 4, and shown to be identical. The inducible gene in H22, an inducible a strain found in our stock cultures and at least one of the inducible genes in H1-0 were linked to the mating type locus. All the inducible genes observed so far were not specific to the mating type in their action.