Restitution of vimba (Vimba vimba, Cyprinidae) in Poland: genetic variability of existing and restored populations

Until 1960s, vimba was found abundantly in Polish rivers and was an important species for the fishing industry. Overfishing, water pollution and dam constructions brought most of the vimba populations close to extinction. We analyzed the genetic variability of several remnant vimba populations in the Vistula and Oder river basins. The 305-bp fragment encompassing the 5′-end of the mtDNA control region was sequenced in a total of 202 wild specimens derived from six populations. Six different haplotypes were distinguished. The population of Barycz River was found to be the most polymorphic and was chosen as a source of breeding material for the vimba restitution program. In the years 2000–2008 fishes from this river were caught, spawned in captivity and released to the same river. The number of fishes returning to the Barycz River spawning grounds was monitored in the following years. In 2008, it was almost 70 times higher than the number of fishes recorded in 2003, when the lowest population size was observed. The haplotype and nucleotide diversities of the restored population were similar to those of the original population.     

Secoiridoid glycosides production by Centaurium maritimum (L.) Fritch hairy root cultures in temporary immersion bioreactor

Secoiridoid glycosides are naturally occurring phytochemicals of great importance for the food and pharmaceutical industry because of their various biological activities. Certain Gentiana and Centaurium species, which are recognized as the most important sources of these compounds, have become critically endangered due to overexploitation. In this study we describe a laboratory-scale approach for further implementation in large-scale production of secoiridoid glycosides, using a hairy root culture system of Centaurium maritimum L. Fritch, an underutilized and phytochemically unexplored species. Hairy roots were induced by Agrobacterium rhizogenes strain A40M70GUS and grown in Erlenmeyer flasks, as well as in RITA^® temporary immersion bioreactors (TIBs). About 2–4 times higher biomass production rate and up to 8 times higher total secoiridoid glycosides production rate were achieved in RITA^® bioreactors. Among the selected hairy root lines, line HR3 cultured in RITA^® TIBs proved to be the most efficient considering both biomass and secoiridoid glycosides production rate.     

The influence of Al3+ ion on porcine pepsin activity in vitro

The in vitro effect of Al³⁺ ions in the concentration range 1.7·10^{? 6} M–8.7·10^{? 3} M on pepsin activity at pH 2, via kinetic parameters and its electrophoretic mobility was evaluated. Kinetic study demonstrated the existence of an activation effect of Al³⁺ at pH 2 on pepsin molecule. Kinetic analysis with respect to concentrations of haemoglobin showed that Al³⁺ ions increase the maximal velocity (V_max) and k_cat values rather than apparent affinity for substrate (K_S) implying the non-competitive nature of activation which indicated that aluminium was a non-essential activator of partial non-competitive type. The values of the equilibrium constants K_S and K_mA for dissociation of corresponding complexes were evaluated as 0.904 ± 0.083 mM and 8.56 ± 0.51 μM, respectively. Dissociation constant K_A, of activator from enzyme-activator complex calculated via kinetic and direct measurement of Al³⁺ binding data, as well as activation constant A₅₀, the activator concentration that gives a rate equal to half at a saturating concentration of activator, were found to be 8.82 ± 0.90 μM, 8.39 ± 0.76 μM, and 8.05 ± 0.48 μM respectively. Native PAGE electrophoresis shows the decrease in electrophoretic mobility of pepsin and confirms modification of the electric charge and conformational changes of pepsin caused by bound Al³⁺ on the pepsin molecule. Al³⁺ induced conformational changes of pepsin were verified by UV-VIS and IR spectra. Moreover, the absence of conformational changes in the haemoglobin molecule in the presence of Al³⁺ ions confirms that the obtained activation is a consequence of conformational changes caused only in the pepsin molecule.     

Reduction and alkylation of peanut allergen isoforms Ara h 2 and Ara h 6; characterization of intermediate- and end products

Conglutins, the major peanut allergens, Ara h 2 and Ara h 6, are highly structured proteins stabilized by multiple disulfide bridges and are stable towards heat-denaturation and digestion. We sought a way to reduce their potent allergenicity in view of the development of immunotherapy for peanut allergy. Isoforms of conglutin were purified, reduced with dithiothreitol and subsequently alkylated with iodoacetamide. The effect of this modification was assessed on protein folding and IgE-binding. We found that all disulfide bridges were reduced and alkylated. As a result, the secondary structure lost α-helix and gained some β-structure content, and the tertiary structure stability was reduced. On a functional level, the modification led to a strongly decreased IgE-binding. Using conditions for limited reduction and alkylation, partially reduced and alkylated proteins were found with rearranged disulfide bridges and, in some cases, intermolecular cross-links were found. Peptide mass finger printing was applied to control progress of the modification reaction and to map novel disulfide bonds. There was no preference for the order in which disulfides were reduced, and disulfide rearrangement occurred in a non-specific way. Only minor differences in kinetics of reduction and alkylation were found between the different conglutin isoforms. We conclude that the peanut conglutins Ara h 2 and Ara h 6 can be chemically modified by reduction and alkylation, such that they substantially unfold and that their allergenic potency decreases.     

Quantification of arabinogalactan proteins during in vitro morphogenesis induced by β-d-glucosyl Yariv reagent in Centaurium erythraea root culture

Arabinogalactan proteins (AGPs) are a family of highly glycosylated cell surface proteins located at the plasma membrane and plant cell wall. AGPs play important roles in plant growth and development. Yariv phenylglycoside (βGlcY), synthetic red-brown dye that specifically binds and precipitates AGPs, has been used for detection and quantification of AGPs in plant tissue. Graded concentrations of βGlcY (0–75 μM) were used to investigate the effect of this synthetic dye on induction of in vitro morphogenesis in Centaurium erythraea root culture on two nutrient media: ½MS and ½MS + IBA 1.0 μM. Regeneration of C. erythraea shoots on root explants was stimulated on both media supplemented with 25 μM βGlcY after 8 weeks in culture. Quantification of AGPs in different tissues of C. erythraea was determinate with single radial diffusion method. This work emphasizes clear effect of βGlcY on induction of morphogenesis in vitro in C. erythraea root culture.     

Phenotypic plasticity in response to an irradiance gradient in <Emphasis Type="Italic">Iris pumila</Emphasis>: adaptive value and evolutionary constraints

Adaptive values of plasticity in Iris pumila leaf traits (morphological: SLA, specific leaf area; anatomical: SD, stomatal density; LT, leaf thickness; VBN, vascular bundle number; SW, sclerenchyma width; CW, cuticle width, and physiological: ChlT, total chlorophyll concentration; ChlA/B, chlorophyll a/b ratio) were tested at three irradiance levels in a growth-room. Siblings from 28 full-sib families from an open dune site and a woodland understory responded similarly to variation in light availability: SLA gradually increased, while anatomical and physiological traits decreased with light reduction. In the Dune population, standardized linear selection gradients were significant for SLA and ChlT at high light, VBN along the entire light gradient, SW at high- and low-, and ChlA/B at low-irradiance. In the Woods population, the significant standardized linear selection gradients were observed for SLA and LT at low- and VBN at both high- and low-irradiance. A significant nonlinear selection gradient was recorded for SD and LT at medium irradiance. Comparisons of the plastic responses to each light quantity with the phenotypes favored by selection in that environments revealed that only an increased SLA value at low light in the Woods population was ecologically significant (adaptive). In the Dune population, SD and VBN entailed plasticity costs at low irradiance, while a cost of homeostasis was recognized for ChlT and ChlA/B at medium light, SD and CW at high- and low-, and SLA at high- and medium-light level. In the shaded population, CW and ChlA/B incurred plasticity costs at high irradiance, while for ChlT plasticity costs appeared under medium- and low-light conditions. In all leaf traits, genetic variation for plasticity was statistically undetectable. Genetic correlations between these traits were mostly insignificant, implying that they possess a capability for relatively independent evolution by natural selection across different light environments.     

Alkaline soda Lake Velika Rusanda (Serbia): the first insight into diatom diversity of this extreme saline lake

Alkaline soda lakes are unique habitats found in specific geographic regions, usually with dry climate. The Carpathian Basin is one of those regions very important for habitat and biodiversity conservation in Europe, with natural soda lakes found in Austria, Hungary and Serbia. In comparison to other two countries from Central Europe, algal biodiversity studies of saline soda lakes in Serbia are scarce. Lake Velika Rusanda has the highest measured salinity of all saline lakes in the Carpathian Basin and there were no reports of its diatom species richness and diversity till now. We conducted 2-year investigation programme to study biodiversity and seasonal dynamics of diatoms in this lake. A total of 27 diatom taxa were found, almost all of them attached to reed and much less in benthos and plankton. Five new diatom species for Serbia were recorded, Craticula halopannonica, Navicymbula pusilla, Hantzschia weyprechtii, Nitzschia thermaloides and Navicula staffordiae. The last mentioned is new for Europe as well. Lake Velika Rusanda is inhabited mostly by alkaliphilous and halophilic diatoms. Since diatoms are used as bioindicators in soda lakes, our results will improve their further application in ecological status assessment of these fragile habitats in the Carpathian Basin.