Functional analysis of the C-terminal propeptide of keratinase from Bacillus licheniformis BBE11-1 and its effect on the production of keratinase in Bacillus subtilis

The keratinase from Bacillus licheniformis BBE11-1 is a serine protease and expressed as a pre-pro-precursor. To produce a mature and active keratinase, the propeptide must be cleaved on the C-terminal via cis or trans. In this study, to enhance the production of keratinase in Bacillus subtilis, single amino acid substitutions, single residue deletions and linkers were introduced at the C-terminus of the propeptide. The results showed that optimizing the residue of cleavage site of propeptide will affect the cleavage efficiency of propeptide, and the mature enzyme yield of Leu(P1)Ala mutant increases 50% compared with the wild-type. In addition, inserting linkers and deleting individual residues at the C-terminal of the propeptide decreases the mature keratinase production. Our results indicated that the primary structure of the C-terminus of propeptide is crucial for the mature keratinase production. Propeptide engineering at C-terminus may be an effective approach to increase the yield of keratinase.     

The Effect of Leaf Age on Gas Exchange and Malate Accumulation in C3-CAM Plant Marrubium Frivaldszkyanum (Lamiaceae)

For the first time the expression of C₃ and CAM in the leaves of different age of Marrubium frivaldszkyanum Boiss, is reported. With increasing leaf age a typical C₃ photosynthesis pattern and high transpiration rate were found. In older leaves a shift to CAM occurred and the 24-h transpiration water loss decreased. A correlation was established between leaf area and accumulation of malate. Water loss at early stages of leaf expansion may be connected with the shift to CAM and the water economy of the whole plant.     

Vertical partitioning between sister species of Rhizopogon fungi on mesic and xeric sites in an interior Douglas‐fir forest

Understanding ectomycorrhizal fungal (EMF) community structure is limited by a lack of taxonomic resolution and autecological information. Rhizopogon vesiculosus and Rhizopogon vinicolor (Basidiomycota) are morphologically and genetically related species. They are dominant members of interior Douglas‐fir (Pseudotsuga menziesii var. glauca) EMF communities, but mechanisms leading to their coexistence are unknown. We investigated the microsite associations and foraging strategy of individual R. vesiculosus and R. vinicolor genets. Mycelia spatial patterns, pervasiveness and root colonization patterns of fungal genets were compared between Rhizopogon species and between xeric and mesic soil moisture regimes. Rhizopogon spp. mycelia were systematically excavated from the soil and identified using microsatellite DNA markers. Rhizopogon vesiculosus mycelia occurred at greater depth, were more spatially pervasive, and colonized more tree roots than R. vinicolor mycelia. Both species were frequently encountered in organic layers and between the interface of organic and mineral horizons. They were particularly abundant within microsites associated with soil moisture retention. The occurrence of R. vesiculosus shifted in the presence of R. vinicolor towards mineral soil horizons, where R. vinicolor was mostly absent. This suggests that competition and foraging strategy may contribute towards the vertical partitioning observed between these species. Rhizopogon vesiculosus and R. vinicolor mycelia systems occurred at greater mean depths and were more pervasive in mesic plots compared with xeric plots. The spatial continuity and number of trees colonized by genets of each species did not significantly differ between soil moisture regimes.     

Food utilization by insects: Interpretation of observed differences between dry weight and energy efficiencies

Over 80% of the values of approximate digestibility (AD), efficiency of conversion of assimilated food to biomass (ECD) and efficiency of conversion of ingested food (ECI) calculated using energy terms are greater than the corresponding dry weight (DW) values, based on data for over 65 species (38 studies; number of comparative values: AD=139, ECD=128 and ECI=169). Largest positive differences (energy > DW values) are 30 (AD, ECD) and 24 (ECI) percentage points and largest negative differences (energy < DW values) are 9 (AD), 11 (ECD) and 8 (ECI) percentage points. These differences generally are least for ECI (71% of the differences fall between 0 and +5 percentage points), and AD (68%), followed by ECD (only 47% fall between 0 and +5), and they may vary with temperature, food and other factors. The differences tend to increase (esp. for ECD and ECI) when comparing later with earlier instars. Energy > DW efficiency values are commonly expected for AD because of the generally greater energy content of food than feces, and for ECD and ECI because of the generally greater energy content of insect biomass than ingested and assimilated food. Deviations from predicted differences in surveyed literature data are discussed in terms of possible methodological sources of error.

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Résumé Plus de 80% des valeurs de la digestibilité approchée (AD), de l'efficacité de la conversion de la nourriture assimilée en biomasse (ECD) et de l'efficacité de la conversion de l'aliment ingéré (ECI), calculées en termes énergétiques, et obtenus à partir de données sur 65 espèces, sont supérieures aux valeurs des poids secs correspondants (DW): 38 études; valeurs comparatives: AD=139, ECD=128, ECI=169. Les plus importantes différences positive (énergie>valuers DW) sont de 30 (AD, ECD) et de 24 (ECI) centièmes (les différences négatives les plus fortes = 9 (AD), 11 (ECD) et 8 (ECI); ces différences sont moindres pour ECI (71% des différences tombent à 0 et +5 centièmes), et AD (68%), suivi de ECD (seulement 47% tombent entre 0 et +5). Ces différences peuvent varier avec la température, l'alimentation et d'autres facteurs; les différences tendent à croître (particulièrement pour ECD et ECI) quant on les compare plus tard avec des stades plus précoces. Energie > aux valeurs d'efficacité DW sont généralement attendues pour AD par suite du contenu énergétique supérieur de l'aliment à celui des excréments, et pour ECD et ECI par suite du contenu énergétique généralement plus élevé pour la biomasse de l'insecte que pour l'aliment ingéré et assimilé. Les écarts par rapport aux différences prédites dans les données de la littérature examinée sont analysées en considérant les sources possibles d'erreurs méthodologiques.

     

The effect of behavioural and morphological plasticity on foraging efficiency in the threespine stickleback (Gasterosteus sp.)

We conducted an experiment to assess the change in foraging efficiency resulting from diet-induced morphological and behavioural plasticity in a species of freshwater, threespine stickleback (Gasterosteus sp.). Different degrees of morphological and behavioural change were induced using two prey items commonly found in the diet of this species, allowing us to estimate the relative importance of each type of plasticity. The purpose of the experiment was twofold. First, earlier work had suggested that diet variability might be an important factor in the evolution of trophic morphological plasticity in sticklebacks. The present results extend this work by revealing the adaptive significance of morphological plasticity. The current experiment also qualitatively assessed the compatibility of the time scale of morphological change with that of the natural resource variability experienced by this species. The results indicate that diet-induced plasticity improves foraging efficiency continuously for up to 72 days of prey exposure. This is probably due in part to plasticity of the external trophic morphology but our results also suggest a complex interplay between morphology and behaviour. The time scale appears to be matched to that of natural diet variability although it is possible that some traits exhibit non-labile plasticity. Our discussion highlights the important distinction between conditions favouring the evolution of labile versus non-labile plasticity. The second objective of the experiment was to determine the relative importance of morphological and behavioural plasticity. Few studies have attempted to quantify the adaptive significance of morphological plasticity and no study to our knowledge has separated the effects of morphological and behavioural plasticity. Our experiment reveals that both behavioural and morphological plasticity are important and it also suggests a dichotomy between the two: behavioural plasticity predominately affects searching efficiency whereas morphological plasticity predominately affects handling efficiency.     

Influence of Nutrient Concentrations on MPN Quantification and Enrichment of Nitrate-Reducing Fe(II)-Oxidizing and Fe(III)-Reducing Bacteria from Littoral Freshwater Lake Sediments

The application of culture-dependent studies to quantify Fe-metabolizing microorganisms from the environment is a necessity, as there are so far no universal functional marker genes for application in culture-independent studies. Media composition can vary between studies, therefore, we determined the effects of three different growth media on the quantification (MPNs) and identity (via cloning and sequencing of dominant DGGE bands) of nitrate-reducing Fe(II)-oxidizers and lactate- or acetate-oxidizing Fe(III)-reducers from a lacustrine sediment: low sulphate freshwater medium (FWM), sterile filtered bicarbonate-buffered lake water (BLW) and a mixture of both (MIX). We consistently found fewer cells in the BLW than in the FWM and the MIX. The DGGE banding patterns of the microbial communities enriched in different media types clustered together according to the e^? donor and acceptor couples and not according to the medium used. Thus, although the medium composition significantly influenced the quantification and thereby conclusions on the abundance and potential significance of the targeted group within the ecosystem, biodiversity assessments through enrichment cultures were less influenced by the medium, but instead were affected by the type and concentration of the e^? donor/acceptor.     

Translocation - A key factor limiting the efficiency of nitrogen fixation in legume nodules

A hypothesis is presented that the availability of water for export of nitrogenous products from legume nodules is a major factor limiting the efficiency of symbiotic nitrogen fixation. Water for export of solutes in the xylem probably depends largely on the import of water and reduced carbon in the phloeum, and one function of respiration may be to dispose of reduced carbon in order to increase the supply of water. A second hypothesis presented is that control of gas diffusion in soybean nodules is largely restricted to the cortex nearby the vascular bundles, thus making possible the linkage of solute balances in xylem and phloem with resistance to diffusion. These concepts are used in a re-examination of literature on manipulations of nodules and nodulated plants such as lowering of light levels, water stress, defoliation, stem girdling, and alteration of oxygen supply. The concept of translocation as a major factor limiting efficiency of symbiotic fixation is consistent with the failure of superior rhizobial isolates to improve N input significantly, and this limitation could also prevent exploitation of superior bacterial symbionts in the future     

Bacterial ecology, antibiotics and selection for virulence

The evolution of antibiotic resistance in bacteria is well known. Here I describe possible mechanisms by which an increased rate of re-colonization of vertebrate guts by microbes caused by antibiotic use could lead to selection for increased virulence in currently mutualistic or benign microbes. The importance of understanding both the source and the frequency of colonization in such mutualisms is stressed and the possible importance of pseudo-vertical transmission in the evolution of these systems is discussed. A number of areas requiring experimental investigation are identified.