Siderophore production by the marine-derived Aureobasidium pullulans and its antimicrobial activity

Over 300 yeast strains isolated from different marine environments were screened for their ability to produce siderophore. Among them, only the yeast strain HN6.2 which was identified to be Aureobasidium pullulans was found to produce high level of the siderophore. Under the optimal conditions, this yeast strain could produce 1.1 mg/ml of the siderophore. The crude siderophore produced by the yeast strain HN6.2 was able to inhibit cell growth of Vibrio anguillarum and Vibrio parahaemolyticus, isolated from the diseased marine animals.     

Characterization and differentiation of 59 strains of Moraxella urethralis from clinical specimens

The biochemical characteristics of 59 strains of Moraxella urethralis from clinical specimens, primarily from urine and the female genital tract, were studied. The characteristics included (i) the inability to acidify carbohydrate substrates, (ii) the ability to produce phenylalanine deaminase, (iii) the ability to reduce nitrite, (iv) the lack of urease activity, and (v) the ability of most strains to alkalinize citrate. A means of differentiating M. urethralis from Moraxella osloensis and Moraxella phenylpyruvica was determined.     

Yarrowia lipolytica: the novel and promising 2-phenylethanol producer

This is the first report on the ability of Yarrowia lipolytica strains to produce 2-phenylethanol (2-PE), which has not been identified for this species to date. 2-PE is a valuable aroma compound of rose-like odor. Its isolation from the other than microbial source—rose petals, is limited by the substrate availability. Thus, this chemical compound constitutes an attractive product for biotechnological conversions. To date, the ability to produce 2-PE has been described for such genera as Saccharomyces sp., Kluyveromyces sp., Geotrichum sp., and Pichia sp. This report provides evidence that Y. lipolytica is a novel 2-PE producer. Moreover, the titers of 2-PE obtained in Y. lipolytica NCYC3825 non-optimized cultures, nearly 2 g/l, are competitive to titers obtained by the other species.     

Cloning of Genes Involved in the Synthesis of Pyrrolnitrin from Pseudomonas fluorescens and Role of Pyrrolnitrin Synthesis in Biological Control of Plant Disease

A soil isolate of Pseudomonas fluorescens (BL915) was shown to be an effective antagonist of Rhizoctonia solani-induced damping-off of cotton. Investigation of the biological basis of this antagonism revealed that the strain produces pyrrolnitrin, a secondary metabolite known to inhibit R. solani and other fungi. Mutants of strain BL915 that did not produce pyrrolnitrin and did not suppress damping-off of cotton by R. solani were generated by exposure to N-methyl-N′ -nitro-N-nitrosoguanidine. A gene region that was capable of restoring pyrrolnitrin production to the non-pyrrolnitrin-producing mutants and of conferring this ability upon two other P. fluorescens strains not otherwise known to produce this compound or to be capable of suppressing damping-off caused by R. solani was isolated from strain BL915. The non-pyrrolnitrin-producing strains (mutants of BL915 and the other two P. fluorescens strains) which synthesized pyrrolnitrin after the introduction of the gene region from strain BL915 were also shown to be equal to strain BL915 in their ability to suppress R. solani-induced damping-off of cotton. These results indicate that we have isolated from P. fluorescens BL915 a gene(s) that has a role in the synthesis of pyrrolnitrin and that the production of this compound has a role in the ability of this strain to control damping-off of cotton by R. solani.     

Promotion of tomato (Lycopersicon esculentum Mill.) plant growth by rhizosphere competent 1-aminocyclopropane-1-carboxylic acid deaminase-producing streptomycete actinomycetes

The ability of streptomycete actinomycetes to promote growth of tomato through the production of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase was evaluated under gnotobiotic and greenhouse conditions. To achieve this, 64 isolates of Streptomyces spp. obtained from a tomato rhizosphere in the United Arab Emirates were initially selected for their ability to produce ACC deaminase as well as indole-3-acetic acid (IAA) and subsequently for their rhizosphere competence as root colonizers. Of the two selected ACC deaminase-producing isolates showing exceptional rhizosphere competence, S. filipinensis no. 15 produced both ACC deaminase and IAA, whilst S. atrovirens no. 26 did not produce IAA. Under greenhouse conditions, the application of S. filipinensis no. 15 or S. atrovirens no. 26 resulted in the reduction of the endogenous levels of ACC, the immediate precursor of ethylene, in both roots and shoots and increased plant growth. Plant growth promotion was most pronounced in the presence of S. filipinensis no. 15 compared to S. atrovirens no. 26. This relative superiority in performance shows the advantage conferred to S. filipinensis no. 15 due to its ability to produce both IAA and ACC deaminase. In comparison, an ACC deaminase-producing isolate of S. albovinaceus no. 41 which was neither rhizosphere-competent nor capable of producing IAA, failed to promote plant growth compared to S. filipinensis no. 15 or S. atrovirens no. 26 although the growth promotion obtained by S. albovinaceus no. 41 was significant compared to control. The application of S. globosus no. 8, which was not rhizosphere-competent and did not produce detectable levels of ACC deaminase or IAA did not promote plant growth. These results indicate the importance of rhizosphere competence. In conclusion I report the production of ACC deaminase by streptomycete actinomycetes and its ability to enhance plant growth through reduction in the in planta levels of endogenous ACC and the consequent lowering of endogenous ethylene levels in plant tissues.     

Vassiljukia,a new colonial rugose coral from the Early Visean (Mississippian) of the Donets Basin (Ukraine) and NW Turkey

The cerioid colonial coral previously described as Lithostrotion columnariformis from the Early Visean of the Donets Basin (Ukraine) is here reattributed to the new genus Vassiljukia. This genus is introduced for colonial amygdalophylloid developing a stable cerioid habitus. It differs from amygdalophylloid proto-colonies by its ability to produce second generation offsets. This ability is proposed here as a definition to differentiate proto-colonial stages from genuine colonies within the rugose corals. Vassiljukia columnariformis is also known from equivalent strata of northwestern Turkey where it occurs with the oldest cerioid Lithostrotion and Ceriodotia. The origin and affinity of Vassiljukia columnariformis within the Amygdalophyllidae are also discussed.     

Host-Specific Pathogenicity and Genome Differences between Inbred Strains of Meloidogyne hapla

Five isolates of M. hapla originating from the Netherlands and California were inbred by sequential transfer of single egg masses to produce six strains. Cytological examination showed that oocytes of these strains underwent meiosis and had n = 16 chromosomes. Strains were tested for ability to infect and to develop on several hosts by in vitro assays. The two strains from California infected tomato roots at a higher rate than those from the Netherlands, but no difference among strains was seen for ability to develop on tomato with or without the resistance gene Mi-1. All strains developed on the common bean cultivar Kentucky Wonder, but strains differed in ability to develop on the nematode-resistant cultivar NemaSnap. Strain-specific differences were also seen in ability to infect and to develop on Solanum bulbocastanum clone SB-22. Strain VW13, derived from nematodes treated with the mutagen EMS, was defective in ability to infect tomato and potato roots in vitro. Comparison of DNA using AFLP markers showed an average of 4% of the bands were polymorphic across the six strains, but no correlation was observed between the geographical origin or virulence and DNA polymorphism pattern.     

Concentrated Cultures of Leuconostoc citrovorum

Two single-strain cultures of Leuconostoc citrovorum were grown in a broth medium with automatic pH control. Culture concentrates were prepared by centrifugally harvesting the cells and resuspending them in 1/50th the original volume in 10% nonfat milk solids. The concentrates were stored in liquid nitrogen until analyzed. The maximum population attainable was approximately equal when cultures were grown at pH 6.0, 6.5, or 7.0 with sodium hydroxide or ammonium hydroxide as the neutralizer. Citrate was required in the growth medium for the cultures to be able to produce diacetyl subsequently in milk. At pH 6.0, the cells reached maximum population and ability to produce diacetyl. Organoleptic analysis by an experienced flavor panel showed a preference for cottage cheese creamed with a creaming mixture prepared with a culture concentrate rather than a normal culture. The culture concentrates maintained their viability and ability to produce diacetyl for at least 30 days when stored in liquid nitrogen.     

New antibiotic-producing Streptomyces TT-strain,generated by electrical fusion of protoplasts

Interspecific electrofusion between protoplasts of multhiomycin-producing Streptomyces antibioticus aAL Ala⁻Leu⁻ and neomycin-producing S. fradiae fHL His⁻Leu⁻ was done. When the concentration of both protoplasts increased to 1 × 10⁹ protoplasts/ml, the frequency of fusion attained was 18.6%. The addition of multithiomycin and neomycin to the regeneration medium was very effective for screening for fusants able to produce new antibiotics. The ability to produce new antibiotics was very unstable in the fusants. After several subcultures, fusants selected as new antibiotic producers (10 strains) lost this ability with one exception (TT-strain). The antibiotic produced by the stable TT-strain clone was purified and characterized to some extent. It was active on a range of Gram-positive bacteria and distinguished from multhiomycin and neomycin by bioautographic analysis.     

There it is! Fusarium pseudograminearum did not lose the fusaristatin gene cluster after all

Fusarium pseudograminearum is a significant pathogen of cereals in arid regions worldwide and has the ability to produce numerous bioactive secondary metabolites. The genome sequences of seven F. pseudograminearum strains have been published and in one of these strains, C5834, we identified an intact gene cluster responsible for biosynthesis of the cyclic lipopeptide fusaristatin A. The high level of sequence identity of the fusaristatin cluster remnant in strains that do not produce fusaristatin suggests that the absence of the cluster evolved once, and subsequently the resulting locus with the cluster fragments became widely dispersed among strains of F. pseudograminearum in Australia. We examined a selection of 99 Australian F. pseudograminearum isolates to determine how widespread the ability to produce fusaristatin A is in F. pseudograminearum. We identified 15 fusaristatin producing strains, all originating from Western Australia. Phylogenetic analyses could not support a division of F. pseudograminearum into fusaristatin producing and nonproducing populations, which could indicate the loss has occurred relatively recent.     

Aflatoxin Biosynthesis Cluster Gene cypA Is Required for G Aflatoxin Formation

Aspergillus flavus isolates produce only aflatoxins B1 and B2, while Aspergillus parasiticus and Aspergillus nomius produce aflatoxins B1, B2, G1, and G2. Sequence comparison of the aflatoxin biosynthesis pathway gene cluster upstream from the polyketide synthase gene, pksA, revealed that A. flavus isolates are missing portions of genes (cypA and norB) predicted to encode, respectively, a cytochrome P450 monooxygenase and an aryl alcohol dehydrogenase. Insertional disruption of cypA in A. parasiticus yielded transformants that lack the ability to produce G aflatoxins but not B aflatoxins. The enzyme encoded by cypA has highest amino acid identity to Gibberella zeae Tri4 (38%), a P450 monooxygenase previously shown to be involved in trichodiene epoxidation. The substrate for CypA may be an intermediate formed by oxidative cleavage of the A ring of O-methylsterigmatocystin by OrdA, the P450 monooxygenase required for formation of aflatoxins B1 and B2.     

Failure of bacterial endotoxin to produce the generalized Shwartzman reaction in the Syrian hamster

The ability of bacterial endotoxin to produce the generalized Shwartzman reaction (GSR) in pregnant and nonpregnant hamsters was investigated. Endotoxins prepared from Escherichia coli O127:B8, Salmonella enteritidis, and S. typhosa 0-901 did not produce the GSR in nonpregnant hamsters. Injection of lead acetate did not make the hamsters susceptible to the GSR producing effects of endotoxin. Endotoxin administered to hamsters on either or both the 14th and 15th day of the 16-day gestation period caused fetal death, but did not provoke the GSR. The immunization of hamsters with boiled suspensions of gram-negative bacteria isolated from hamster feces did not protect against the GSR produced in pregnant hamsters by the injection of the antimitotic drug colchicine late in the gestation period. It appeared that colchicine was acting to produce the GSR by a mechanism other than the release of endogenous endotoxin through the damaged intestinal wall. Ascitic fluid, amniotic fluid, and serum obtained from pregnant hamsters developing the GSR after the administration of colchicine did not provoke the GSR in other pregnant hamsters.     

Detection and quantification of patulin and griseofulvin by high pressure liquid chromatography in different strains of Penicillium griseofulvum Dierckx

Patulin and griseofulvin production by twelve strains ofPenicillium griseofulvum Dierckx, eleven of which were isolated from pistachio (Pistacia vera) nuts and the other was supplied by the Spanish Collection of Type Culture, was investigated. Six strains of the eleven isolated had ability to produce patulin and griseofulvin in Yes medium. All the strains studied had no ability to produce patulin in Wickerham medium. Griseofulvin production was significant in both media but higher in Wickerham. These metabolites were separated and determined in the chloroform extracts of cultures by high performance liquid chromatography with ultraviolet detection. The best conditions were: acetonitrile — water (45∶55) as mobile phase, a flow rate of 2.0 mL/min and a μ Bondapack C₁₈ column.     

Isolation of natural cultures of anaerobic fungi and indigenously associated methanogens from herbivores and their bioconversion of lignocellulosic materials to methane

This study aimed to obtain natural cultures of anaerobic fungi and their indigenously associated methanogens from herbivores and investigate their ability to degrade lignocelluloses to methane. Eight natural cultures were obtained by Hungate roll tube technique. The fungi were identified as belonging to Piromyces, Anaeromyces and Neocallimastix respectively by microscopy, and the methanogens as Methanobrevibacter spp. by 16S rRNA gene sequencing. In vitro studies with rice straw showed that these cultures degraded 33.5-48.3% substrate and produced 0.33-0.84 mmol/(100 ml culture) methane. Two cultures were further selected for their ability to degrade different lignocellulosic materials and could produce 0.38-1.27 mmol/(100 ml culture) methane. When methanogens were inhibited, the lignocellulose-degrading ability of cultures significantly reduced. In conclusion, natural cultures of anaerobic fungi with indigenously associated methanogens with high fiber degradation ability were obtained, and these cultures may have the potential in industrial use in lignocelluloses degradation and methane production.     

Isolation and characteristics of the lactose-fermenting yeasts Candida kefyr

Screening of lactose-fermenting yeast strains has been conducted among 162 strains isolated from various plants and 28 strains isolated from cheese. Four yeast strains fermented lactose and were identified as Candida kefyr. The specific β-galactoside activity of the studied strains was 1501–2113 IU/g dry biomass. The ability of strains C. kefyr C24 and C30 to produce ethanol from lactose was significantly inhibited by the increase in substrate concentration (100 g/L).     

Control of yeast cell type by the mating type locus: I. Identification and control of expression of the a-specific gene BAR1

The MATα allele of the yeast mating type locus confers the α mating phenotype and contains two complementation groups, MATα1 and MATα2. The α1–α2 hypothesis proposes that MATα1 is a positive regulator of α-specific genes and that MATα2 is a negative regulator of a-specific genes. According to this hypothesis, matα2 mutants, which are defective in mating and in production of extracellular α-factor, express both a-specific functions (because they lack MATα2 product) and α-specific functions (because they contain MATα1 product). Failure to produce extracellular α-factor results from antagonism between these functions; in particular, because α-factor (an α-specific function) is degraded by an a-specific function. If this view is correct, matα2 mutants should acquire the ability to produce α-factor if they also carry a defect in the gene(s) responsible for α-factor degradation. We have isolated a derivative of a matα2 mutant that produces α-factor and have characterized the suppressor mutation in this strain. (1) This strain carries a mutation (bar1-1) tightly linked to HIS6 (on chromosome IX) that allows matα2 mutants to produce α-factor. (2) It does not allow matα1 mutants to produce α-factor. (3) Haploids of the a mating type bearing the bar1-1 mutation still mate, but are unable to act as a barrier to the diffusion of α-factor. MATa bar1-1 cells display increased sensitivity to α-factor. (4) A mutation (sst1?2) that causes increased sensitivity to α-factor is allelic to bar1-1 and also allows α-factor synthesis by matα2 mutants. The ability of matα2 bar1 double mutants to produce extracellular α-factor indicates that matα2 mutants do produce α-factor but that it is degraded by the Barrier function. These results suggest that BAR1 is normally expressed only in a cells, and is negatively regulated in α cells by the MATα2 product.     

Male production by workers in the polygynous ant Prolasius advenus

The ability of workers to produce male individuals is reported here for the first time in a species of the formicine ant genus Prolasius. We show that Prolasius advenus workers possess ovaries and demonstrate that they are able to produce adult males in queenless colonies. We also experimentally tested the influence of queen volatiles on the level of worker reproduction. Workers produced fewer eggs in treatments where they could perceive odors from queens. Some volatile compounds emitted by queens may thus have a signaling or inhibitory effect on worker reproduction. This effect of queen presence did not entirely stop worker reproduction, however, as adult males still emerged under these conditions. Worker-produced males were absent only in treatments with the physical presence of queens. Dissections of workers collected from queenright nests in the field revealed signs of egg-laying activity in more than half of individuals. Together, these results suggest that in nature P. advenus workers produce males at least in orphaned colonies or in situations where the physical presence of queens is limited.     

CHEMICAL RESTORATION IN NITELLA : II. RESTORATIVE ACTION OF BLOOD

Cells of Nitella exposed to distilled water lose their ability to produce action currents and to distinguish electrically between sodium and potassium. This ability was quickly restored by exposure to blood plasma deprived of calcium. Human blood and that of the cat, calf, and sheep gave essentially the same results. The active agents appear to be organic substances.     

Interference in hybrid clone selection caused by Mycoplasma hyorhinis infection

Mycoplasma hyorhinis strains were isolated from Chinese hamster DON cells which lacked the ability to produce hybrid colonies in HAT medium. The mycoplasma isolates were virtually devoid of HGPRT activity in vivo and in vitro in the presence of excess co-enzyme, phosphoribosylpyrophosphate. Deliberate infection of mycoplasma-free cells caused no alterations in the HGPRT^? and TK^? phenotypes of the cells. Heterokaryon formation with infected cells was normal and the failure to produce hybrid colonies resulted from depletion, by nucleoside phosphorylase activity, of exogenous thymidine required for rescue of hybrid cells in HAT medium. Increasing the thymidine concentration and repeatedly replenishing HAT medium permitted hybrid clone formation.