Fermentation of fructans by epiphytic lactic acid bacteria

A total of 712 strains of lactic acid bacteria isolated from forage grasses were studied for their ability to ferment fructans of phlein- as well as inulin-type. Only 16 strains utilized phlein and eight of these also fermented inulin. They were identified as Lactobacillus paracasei subsp. paracasei, Lact. plantarum, Lact. brevis and Pediococcus pentosaceus . In the species Lact. paracasei subsp. paracasei , all strains gave positive results, whereas the other positive strains possessed unique properties within their own species. In all but two cases (strains of the species Lact. plantarum ), the phlein was more intensively fermented than the inulin, as indicated by a lower pH and a higher lactic acid concentration. On the basis of the outcome of this study it seems worthwhile to inoculate grasses of low sugar content before ensiling with an active strain that can ferment fructans.     

Production of phenolics by immobilized cells of the lichen Pseudevernia furfuracea: the role of epiphytic bacteria

Immobilized lichen cells from the thalli of the lichen Pseudevernia furfuracea, supplied with acetate as the only source of carbon, continuously produced phenolic substances, atranorin and physodic acid, over 23 days. Epiphytic bacteria associated with the lichen thallus grew actively, probably using both acetate and reduced compounds supplied by lichen cells, since their active growth was avoided by including 10 microM 3,3'-dichlorophenyl-1,1'dimethylurea in the bath solution. Penicillin largely impeded the growth of epiphytic bacteria and decreased phenolic production, which was recovered only at the end of the experimental period, just when the bacteria started a slow, but active growth. We suggest the cooperation of epiphytic bacteria in the biosynthesis of both atranotrin and physodic acid.     

Two non-identical heads of myosin molecules

Four different preparations of skeletal subfragment-1, denoted in this report as S1(Aa), S1(Ab), S1(Ba), and S1(Bb), and two different preparations of cardiac subfragment-1, denoted as S1(A) and S1(B), were obtained as described in our recent report (J. Biochem. 97, 965, 1985). (i) The four preparations were obtained from chicken breast myosin trinitrophenylated with 2,4,6-trinitrobenzene sulfonate in the absence of inorganic pyrophosphate (-PPi), and they were all shown to be trinitrophenylated. Addition of PPi caused change in the absorption spectra of trinitrophenyl(TNP)-S1(Aa) and TNP-S1(Ba), but not in those of TNP-S1(Ab) and TNP-S1(Bb). (ii) The two preparations of S1 were obtained from cardiac myosin trinitrophenylated either in the absence (-) or presence (+) of PPi. S1(B) was trinitrophenylated, whereas S1(A) was not. Specifically emphasized is the observation that the yield of cardiac S1(A) was practically equal to that of cardiac S1(B). On the basis of these results, we propose the hypothesis of "two iso-myosins with non-identical heads," which is essentially a combination of the hypothesis of isoenzymes and that of non-identical heads.     

Assessment of atrazine decontamination by epiphytic root bacteria isolated from emergent hydrophytes

Aim of the study was to identify atrazine remediating bacteria that can potentially succeed in situ where they encounter varied environmental conditions. Three epiphytic root bacteria, genus Pseudomonas and Arthrobacter, were isolated from rhizoplanes of hydrophytes Acorus calamus, Typha latifolia, and Phragmites karka. Potential of these strains to decontaminate environmentally relevant concentrations of atrazine was determined in liquid atrazine medium (LAM) and Luria-Bertani (LB) medium at varying pH and temperature. There was an increase in decontamination by the strains with time upon exposure to 2.5 to 10 mg l^?1 atrazine over a period of 15 days, notably, in both minimal and nutrient-rich media. Growth in terms of O.D.₆₀₀ and biomass determined during the same period also showed a corresponding surge. Pseudomonas sp. strain AACB mitigated atrazine in a wide range of pH (5 to 8). Pseudomonas sp. strains AACB and TTLB decontaminated >?62% atrazine at 10 °C. All the strains exhibited plant growth–promoting traits in vitro, reported for the first time in the presence of atrazine. Strain AACB exhibits the novel trait of atrazine decontamination under harsh environmental conditions mimicked in lab. Strains isolated in the present study promise success in in situ remediation. Bioreactors and water treatment plants can be designed comprising the hydrophytes and the strains inoculated into their rhizospheres to improve efficacy of the treatment. They can be used to study plant-bacterium mutualistic symbiosis or other interactions occurring during atrazine mitigation.     

Production of<Emphasis Type="Italic">N</Emphasis>-acylhomoserine lactone signal molecules by gram-negative soil-borne and plant-associated bacteria

Quorum-sensing control mediated by N-acylhomoserine lactone (AHL) signal molecules has been established as a key feature in the regulation of various metabolic traits in many bacteria. Approximately 300 strains representing 6 genera and 18 species of soil-borne and plant-associated Gram-negative bacteria isolated in various regions of the former USSR using two reporter systems were screened for AHL production. The production was observed in 17.5% of the screened bacterial strains. Positive response was detected in all of the 14 tested strains of Erwinia herbicola, in 41 of the 239 strains of Pseudomonas species; in all 5 strains of Xanthomonas ampelina, X. campestris pv. malvacearum, pv. translucens, pv. vesicatoria and in one strain of Pantoea stewartii. AHL assay of 41 strains of X. maltophilia (syn. Stenotrophomonas maltophilia) isolated from soils with Chromobacterium violaceum reporter has revealed no strains synthesizing these signal molecules; 26 strains analyzed with Agrobacterium tumefaciens reporter showed the same result.     

Production of cell–cell signalling molecules by bacteria isolated from human chronic wounds

Aim: To (i) identify chronic wound bacteria and to test their ability to produce acyl‐homoserine‐lactones (AHLs) and autoinducer‐2 (AI‐2) cell–cell signalling molecules and (ii) determine whether chronic wound debridement samples might contain these molecules. Methods and Results: Partial 16S rRNA gene sequencing revealed the identity of 46 chronic wound strains belonging to nine genera. Using bio‐reporter assays, 69·6% of the chronic wound strains were inferred to produce AI‐2, while 19·6% were inferred to produce AHL molecules. At least one strain from every genus, except those belonging to the genera Acinetobacter and Pseudomonas, were indicated to produce AI‐2. Production of AI‐2 in batch cultures was growth‐phase dependent. Cross‐feeding assays demonstrated that AHLs were produced by Acinetobacter spp., Pseudomonas aeruginosa and Serratia marcescens. Independent from studies of the bacterial species isolated from wounds, AHL and/or AI‐2 signalling molecules were detected in 21 of 30 debridement samples of unknown microbial composition. Conclusion: Chronic wound bacteria produce cell–cell signalling molecules. Based on our findings, we hypothesize that resident species generally produce AI‐2 molecules, and aggressive transient species associated with chronic wounds typically produce AHLs. Both these classes of cell–cell signals are indicated to be present in human chronic wounds. Significance and Impact of the Study: Interbacterial cell–cell signalling may be an important factor influencing wound development and if this is the case, the presence of AHLs and AI‐2 could be used as a predictor of wound severity. Manipulation of cell–cell signalling may provide a novel strategy for improving wound healing.     

Growth of epiphytic and soil yeasts on wheat seedlings

Colonization of wheat seedlings by epiphytic (Rhodotorula glutinis) and soil (Lipomyces starkeyi) yeasts was studied by scanning electron microscopy. Epiphytic yeast cells dominated on the plant surface. Soil yeast cells were randomly distributed among both the zones of a seedling and the particles of an inorganic substrate. It has been found that epiphytic yeast strains can readily grow on the surface of a plant.     

Antibiotic activity of epiphytic bacteria isolated from intertidal seaweeds

A survey of antibiotic-producing bacteria from the microbial flora attached to seaweeds and the study of their antibiotic capacities were carried out. From 5 species of green and brown marine algae, 224 bacterial strains were isolated and tested for antibiotic production. A total of 38 strains displayed antibiotic activity, withEnteromorpha intestinalis being the source of the highest number of producer strains. All epiphytic bacteria with antibiotic activity were assigned to thePseudomonas-Alteromonas group. Antagonism assays among the isolates demonstrated that each producer strain inhibits the growth of the other producers, as well as of some nonproducer strains also isolated from seaweeds. Likewise, an autoinhibitory effect was observed in all antibiotic-producing strains. Antibacterial spectra of all the strains include activity againstStaphylococcus, Alcaligenes, Pseudomonas, Vibrio, Pasteurella, andAchromobacter. A preliminary characterization of the antibiotic substances produced by these epiphytic bacteria demonstrated that they are low molecular weight compounds, thermolabile, and anionic and are not affected by proteolytic enzymes. The role that these inhibitory substances can play in the natural environment is discussed.     

Metabolic footprint of epiphytic bacteria on Arabidopsis thaliana leaves

The phyllosphere, which is defined as the parts of terrestrial plants above the ground, is a large habitat for different microorganisms that show a high extent of adaption to their environment. A number of hypotheses were generated by culture-independent functional genomics studies to explain the competitiveness of specialized bacteria in the phyllosphere. In contrast, in situ data at the metabolome level as a function of bacterial colonization are lacking. Here, we aimed to obtain new insights into the metabolic interplay between host and epiphytes upon colonization of Arabidopsis thaliana leaves in a controlled laboratory setting using environmental metabolomics approaches. Quantitative nuclear magnetic resonance (NMR) and imaging high-resolution mass spectrometry (IMS) methods were used to identify Arabidopsis leaf surface compounds and their possible involvement in the epiphytic lifestyle by relative changes in compound pools. The dominant carbohydrates on the leaf surfaces were sucrose, fructose and glucose. These sugars were significantly and specifically altered after epiphytic leaf colonization by the organoheterotroph Sphingomonas melonis or the phytopathogen Pseudomonas syringae pv. tomato, but only to a minor extent by the methylotroph Methylobacterium extorquens. In addition to carbohydrates, IMS revealed surprising alterations in arginine metabolism and phytoalexin biosynthesis that were dependent on the presence of bacteria, which might reflect the consequences of bacterial activity and the recognition of not only pathogens but also commensals by the plant. These results highlight the power of environmental metabolomics to aid in elucidating the molecular basis underlying plant–epiphyte interactions in situ.     

Binding of extracellular matrix molecules by probiotic bacteria

AIMS: The aim of this study was to investigate extracellular matrix (ECM) and mucin binding of selected bacterial isolates with probiotic features in comparison with commercially used probiotic bacteria. METHODS AND RESULTS: ECM molecules were immobilized in microtitre plates (mucin and fetuin) or on the surface of latex beads. Porcine mucin was bound by all 13 probiotic strains tested with important inter-strain differences; however, fetuin binding was similar (weak) for all 14 strains tested. Strongly positive (three) binding of bovine fibrinogen was expressed by strains from fermented food (Lactobacillus rhamnosus GG, L. casei Shirota and L. johnsonii La1) as well as by L. casei L.c., Lactobacillus sp. 2I3 and by L. plantarum LP. The other strains expressed moderate (2) or weakly positive (1) binding of bovine fibrinogen. Strongly positive (3) binding of porcine fibronectin was observed only with two strains; however, all other strains also bound this molecule. Bovine lactoferrin was bound to a higher extent than transferrins. SIGNIFICANCE AND IMPACT OF THE STUDY: Some animal strains (at least L. casei L.c. and Lactobacillus sp. 2I3) are comparable with the commercially used strains with respect to their ECM binding ability. As this feature is important for probiotic bacteria to be able to colonize intestine, these strains should be considered for their wider use in fermented feed (or probiotic preparations) for animals.     

Production of ethanol by thermophilic bacteria

Doubts about the viability of using classical yeast fermentations to produce ethanol have led in recent years to a search for alternative microbes capable of ethanol production. Among the front-runners for consideration are the thermophilic bacteria and this article shows how close these extraordinary microbes have come to replacing yeast in ethanol production.     

The role of indoleacetaldehyde in IAA production from tryptophan by plants and by their epiphytic bacteria

Tryptophan, tryptamine, or indolepyruvic acid were applied to 2 systems: a bacterial (pea stem sections containing the epiphytic bacteria) and a plant system (pea stem sections under sterile conditions). In the plant system, the production of indoleacetic acid and indoleethanol (tryptophol) from each applied indole derivative is clearly reduced by the aldehyde reagents bisulfite and dimedon, respectively. Indoleacetaldehyde is chromatographically detected after alkaline liberation from its bisulfite addition product. In the bacterial system, the production of indoleacetic acid and indoleethanol is likewise reduced by bisulfite and dimedon. However, after tryptophan or tryptamine application, we could not detect indoleacetaldehyde in the described way. In one case only, namely tryptamine application to the bacterial system, indoleethanol production (contrary to indoleacetic acid production) is scarcely reduced by the aldehyde reagents. This indicates a bacterial pathway tryptamine → indoleethanol which bypasses indoleacetaldehyde.     

二氧化硫对小麦的氧化胁迫及其某些信号分子的调节

通过在密闭的培养箱中一次性通入不同体积浓度的SO₂气体,研究了小麦幼苗超氧自由基O2-含量和3种抗氧化酶活性的变化,探讨了信号分子水杨酸、乙烯和过氧化氢对SO₂氧化胁迫的调节作用.结果表明,当通入10和40 μl·L^-1 SO₂时,小麦叶片中O2-含量递增,过氧化物酶(POD)和过氧化氢酶(CAT)活性增强,但超氧化物歧化酶(SOD)活性降低.当SO₂浓度达到50 μl·L^-1时,POD和CAT活性也开始降低,此时叶片尖端出现坏死,叶片绿色部位滋生大量真菌.用1 mmol·L^-1水杨酸(SA)(pH6.5)浸泡小麦干种子6 h,或用10 mmol·L^-1 H₂O₂浸泡幼苗,O2-含量低于对照植株,而3种酶的活性高于对照植株,均可有效地减轻SO₂的氧化胁迫.在SO₂熏蒸下,乙烯显著抑制3种酶的活力,提高O2-的形成速率.SA与乙烯同时使用时,SA几乎完全消除了乙烯的负面作用.     

Detection of quorum-sensing-related molecules in <Emphasis Type="Italic">Vibrio scophthalmi</Emphasis>

Background  

Cell-to-cell communication (also referred to as quorum sensing) based on N-acyl-homoserine lactones (AHLs) is a widespread response to environmental change in Gram-negative bacteria. AHLs seem to be highly variable, both in terms of the acyl chain length and in the chemical structure of the radicals. Another quorum sensing pathway, the autoinducer-2-based system, is present both in Gram-positive and Gram-negative bacteria. In this study the presence of signal molecules belonging to both quorum sensing signalling pathways was analysed in the marine symbiotic species Vibrio scophthalmi.     

Unshadowed myosin molecules: STEM mass-maps of myosin heads.

Myosin molecules were directly visualized without heavy metal shadowing by scanning transmission electron microscopy (STEM) under low dose conditions. The general appearance and dimensions of heavy metal-free molecules were similar to those of shadowed myosin, either after freeze-drying without or air-drying with glycerol. Two characteristic configurations of myosin head regions were found, a first type showing two pear-shaped heads with narrow necks and a second type showing two heads connected by an extra mass in the central regulatory domain where the light chains are located. The mass of the latter type (mol. wt. = 265 +/- 39 kd) is in excellent accordance with biochemical data whereas the mass of the first type is somewhat lower (mol. wt. 219 +/- 44 kd).