Miroslava Hazuchová, Daniela Chmelová and Miroslav Ondrejovič
The lignocellulolytic enzymes are routinely produced by submerged fermentation using lignocellulosic material, but for more effective production, it would be suitable to precede the production phase on the lignocellulose by propagation phase in the nutrition medium suitable for growth of the fungi. Therefore, the aim of this study was to increase the laccase production by the white-rot fungus Pleurotus ostreatus by two-step cultivation strategy. In the first step, propagation medium was optimized for the maximal biomass growth, the second step included the laccase production by produced fungal biomass in media with the selected lignocellulosic material (pine sawdust, alfalfa steam and corn straw). From our experiments, parameters such as glucose concentration, yeast extract concentration and pH of propagation medium were selected as key factors affecting growth of P. ostreatus. The optimal conditions of propagation medium for maximal fungal growth determined by response surface methodology were: glucose concentration 102.68 g/L, yeast extract concentration 43.65 g/L and pH of propagation medium 7.24. These values were experimentally verified and used statistical model of biomass production prediction was appropriate adjusted. Thus prepared fungal biomass produced in the media with lignocellulose approximately 9-16 times higher concentrations of the laccase in 3 times shorter time than the fungal biomass without propagation phase in optimized propagation medium.
In this study, the phytotoxicity of disinfectants prepared on the base of sodium hypochlorite was determined. For our tests two commercial products, Savo and Dom Amor, as well as 10% NaClO solution were used. While Savo contained only NaClO, Dom Amor contained NaClO and earthworm enzymes. Products on the base of NaClO are used in households for cleaning and disinfection of floors, furniture, sanitary and kitchen equipment. Savo may be used for the disinfection of drinking waters as well. Products with NaClO are also used for bacteria, algae and pathogens reduction in irrigation waters. As a subject, young seedlings of mustard Sinapis alba L. were used for the study of chronic toxicity. The observed parameters of the inhibition of roots and shoots growth, dry (DM) and fresh (FM) mass as well as photosynthetic pigments production (chlorophyll a, b, carotenoids) and water content in the plants were determined. The results point out that Dom Amor was the most toxic for S. alba seedlings growth and the rank order of the FAC contents for both plant parts was arranged as: Dom Amor > Savo > NaClO. All disinfectants reduced the DM and FM of roots; however, they stimulated biomass production in the shoots. On the basis of the obtained results it could be concluded, that disinfectants stimulated photosynthetic pigments production and reduced water content mainly in the roots. Dom Amor did not significantly reduced the water content in the shoots and for this parameter the following rank orders of inhibition for roots and shoots could be arranged as NaClO > Dom Amor > Savo and NaClO > Savo > Dom Amor, respectively. All commercial products increased chlorophyll a (Chla) and the carotenoids (Car) content in the shoots. As significant increase was confirmed first for Chla whose content in the presence of NaClO at concentration 24 mL/L overextended that in the control by 3.5 times. The rank orders of stimulation for Chla and Car were NaClO > Savo > Dom Amor and Dom Amor > NaClO > Savo, respectively.
Alexandra Šimonovičová, Sanja Nosalj, Alžbeta Takáčová, Tomáš Mackuľak, Karol Jesenák and Slavomír Čerňanský
Four wild type strains of A. niger were collected from soil and stream sediments representing environments with variable level of As, Sb, Al, Fe, Cd, Cu, and Zn contamination. Banská Štiavnica-Šobov (S), Pezinok-Kolársky vrch (P) and Slovinky (Sl) represent contaminated localities. Locality Gabčíkovo (G) was as a control site. The influence of toxic elements in these substrates on fungal growth, colony size, enzymatic activity, production of organic acids and their pelletization in water suspensions with montmorillonite was studied. The aim of our study was to find out how the wild type strains from (contaminated) environment will behave in different model solutions. We also wanted to add some new information in this area of study, because that there is some gap in the available knowledge.
Monika Bardáčová, Yevheniia Konotop, Zuzana Gregorová, Miroslav Horník, Jana Moravčíková, Ján Kraic and Ildikó Matušíková
Cadmium is a serious environmental pollutant and its uptake by plant represents a serious health risk. Uptake, accumulation as well as sensitivity of soybean plants to metals have been shown to vary with genotype, while the dynamics of this uptake has rarely been studied. Here we studied the uptake and accumulation of Cd2+ ions in different parts of soybean plants of four cultivars Moravians, Gallec, Kent and Cardiff. The plants at early developmental stage were immersed in Hoagland nutrient solution in the presence or absence of 50 mg.L−1 and the isotope of 109Cd2+ to monitor its accumulation continuously at 24 h intervals for 10 days. Our results showed that the uptake rate varied among the cultivars, being the highest in roots of the cv. Moravians and the lowest in the cv. Gallec. We also observed a non-even distribution of radioactivity within the entire plants of individual cultivars. The most of Cd2+ isotope was translocated into primary leaves and leaves in the cvs. Kent and Moravians; on the contrary, relatively less in the cvs. Cardiff and Gallec. The results were fitted with genetic potential, growth as well as defense parameters such as proline accumulation. Combining uptake dynamics and biochemical data are indicative for different tolerance strategies of soybeans.
Farida Y. Achmadulina, Rustem K. Zakirov, Elena S. Balymova, Vera Denisova, Taťjána Brovdyová, Josef Trögl and Martin Neruda
Activated sludge biocenoses were compared on waste-water treatment plants in the city of Kazan, Russian Federation and the city of Teplice, Czech Republic. Based on Palia-Kovnatski index, Acanthamoeba in Kazan, Epistylis in Teplice, and Acanthamoeba and Centropyxis were dominant genera in both plants. The major subdominant generas identified were Arcella, Opercularia and Aspidisca. This indicates high nitrification ability, high water purification potential and matured activated sludge. Chemical composition of the waste-water was identified as the main factor determining the sludge biocenoses diversity. Higher sludge biodiversity (Shannon, Margalef, and Sorensen indexes) was found in Kazan corresponding to more concentrated inflow water.
Nikola Š Šipošová, Veronika Liptáková, Simona Kvasnová, Petra Kosorínová and Peter Pristaš
Multiple metallotolerant bacterial strains were isolated from soil and drainage water samples collected from three industrially heavy metals polluted areas in Slovakia. Obtained bacterial isolates were identified using MALDI-TOF mass spectrometry and bacterial isolates that belonged to the Acinetobacter genus were subjected for the further study. A. calcoaceticus was found to be prevalent species among analyzed Acinetobacter spp. strains, followed by A. lwoffii and A. johnsonii. A. calcoaceticus strains exhibited higher minimum inhibitory concentration to Mn, Zn, and Cu cations compared to A. lwoffii and A. johnsonii. On the other hand minimum inhibitory concentration to Co and Ni were identical in all Acinetobacter spp. isolates. Genetic analyses demonstrated multiple plasmids presence in A. lwoffii and A. johnsonii but not in A. calcoaceticus. Using ERIC-PCR the presence of two different genotypes of A. calcoaceticus was detected in heavy metal polluted environments in Slovakia.
Daniela Jamrichová, Lenka Tišáková, Veronika Jarábková and Andrej Godány
Production of recombinant proteins in Escherichia coli expression systems has shown many advantages, as well as disadvantages, especially for biotechnological and other down-stream applications. The choice of an appropriate vector depends on the gene, to be cloned for purification procedures and other analyses. Selection of a suitable production strain plays an important role in the preparation of recombinant proteins. The main criteria for the selection of the host organism are the properties of the recombinant produced protein, its subsequent use and the total amount desired. The most common problems in eukaryotic gene expression and recombinant proteins purification are, for instance, post-translational modifications, formation of disulphide bonds, or inclusion bodies. Obtaining a purified protein is a key step enabling further characterization of its role in the biological system. Moreover, methods of protein purification have been developed in parallel with the discovery of proteins and the need for their studies and applications. After protein purification, and also between the individual purification steps, it is necessary to test protein stability under different conditions over time. Shortly, all the essential points have been briefly discussed, which could be encountered during production and purification of a recombinant protein of interest, especially from eukaryotic source and expressed heterogeneously in prokaryotic production system.
Denisa Partelová, Klára Kuglerová, Yevheniia Konotop, Miroslav Horník, Juraj Lesný, Marcela Gubišová, Jozef Gubiš, Peter Kováč and Ildikó Matušíková
The current findings show that positron emission tomography (PET), primarily developed for medical diagnostic imaging, can be applied in plant studies to analyze the transport and allocation of wide range of compounds labelled with positronemitting radioisotopes. This work is focused on PET analysis of the uptake and transport of 2-deoxy-2-fluoro[18F]-D-glucose (2-[18F]FDG), as a model of photoassimilates, in tissues of giant reed (Arundo donax L. var. versicolor) as a potential energy crop. The absorption of 2-[18F]FDG and its subsequent transport in plant tissues were evaluated in both acropetal and basipetal direction as well. Visualization and quantification of the uptake and transport of 2-[18F]FDG in plants immersed with the root system into a 2-[18F]FDG solution revealed a significant accumulation of 18F radioactivity in the roots. The transport rate in plants was increased in the order of plant exposure through: stem > mechanically damaged root system > intact root system. PET analysis in basipetal direction, when the plant was immersed into the 2-[18F]FDG solution with the cut area of the leaf of whole plant, showed minimal translocation of 2-[18F]FDG into the other plant parts. The PET results were verified by measuring the accumulated radioactivity of 18F by direct gamma-spectrometry.
Jana Moravčíková, Ildikó Matušíková, Peter Nemeček, Alžbeta Blehová, Želmíra Balážová, Zdenka Gálová, Patrik Mészáros and Ján Kraic
Acceptance of genetically modified plants is restricted in EU by legislation, while the attitude of public is not favourable as well. Surveys show that knowledge about GM plants is getting increased. Newly developed strategies on GM safety for environment can be a crucial aspect for the (partial) acceptance in future. GM trees as non-edible plants might appear as more admissible, however, are relatively rarely discussed. We performed a comparative survey on knowledge and perception of GM forest trees among students at four Slovak universities. We also compared their responses between as well as with the outcome of similar cross-country survey in frames of the COST Action FP0905. The results point to very similar attitude of Slovak students when compared with students from other countries, no significant difference between responses of males and females, but also influence of age as well as orientation of their study (natural sciences vs. economy) on view of GM tree safety and placing on the market.
Ottó Dóka, Andrea Brunori, Rezső Schmidt, Dane Bicanic and György Végvári
A relatively novel approach for easy and quick determination of rutin in buckwheat grain is suggested. The rutin content of the grain in seven common buckwheat (Fagopyrum esculentum) and six Tartary buckwheat (Fagopyrum tataricum) varieties was investigated by means of UV photoacoustic spectroscopy and HPLC as reference method. The lowest content was found in ‘Botan’ and ‘Bamby’ varieties, while the highest values were obtained in the variety ‘Emka’. Rutin content in grain of all Tartary buckwheat varieties was two orders of magnitude higher than in the other varieties. Rutin content in F. esculentum ranges between 9 and 36 mg/100 g dry weight as compared to 921 to 2 132 mg/100 g dry weight in F. tataricum. The UV photoacoustic spectroscopy data show rather good correlations of R2=0.977 and R2=0.980 with values obtained by HPLC data for all measured samples. Therefore, UV photoacoustic spectroscopy can be a cheap and quick method for determining rutin content in buckwheat grain.