Pattern of antioxidant enzyme activities and hydrogen peroxide content during developmental stages of rhizogenesis from hypocotyl explants of Mesembryanthemum crystallinum L.

KEY MESSAGE: H(2)O(2)is necessary to elicit rhizogenic action of auxin. Activities of specific catalase and manganese superoxide dismutase forms mark roots development. ABSTRACT: Hypocotyl explants of Mesembryanthemum crystallinum regenerated roots on medium containing 2,4-dichlorophenoxyacetic acid. Explants became competent to respond to the rhizogenic action of auxin on day 3 of culture, when hydrogen peroxide content in cultured tissue was the highest. l-Ascorbic acid added to the medium at 5 μM lowered the H(2)O(2) level, inhibited rhizogenesis and induced non-regenerative callus, suggesting that certain level of H(2)O(2) is required to promote root initiation. Coincident with the onset of rhizogenic determination, meristemoids formed at the periphery of the hypocotyl stele and the activity of the manganese form of superoxide dismutase, MnSOD-2 was induced. Once induced, MnSOD-2 activity was maintained through the post-determination phase of rooting, involving root growth. MnSOD-2 activity was not found in non-rhizogenic explants maintained in the presence of AA. Analyses of the maximum photochemical efficiency of photosystem II and the oxygen uptake rate revealed that the explants were metabolically arrested during the predetermination stage of rhizogenesis. Respiratory and photosynthetic rates were high during root elongation and maturation. Changes in catalase and peroxidase activities correlated with fluctuations of endogenous H(2)O(2) content throughout rhizogenic culture. Expression of a specific CAT-2 form accompanied the post-determination stage of rooting and a high rate of carbohydrate metabolism during root growth. On the other hand, the occurrence of MnSOD-2 activity did not depend on the metabolic status of explants. The expression of MnSOD-2 activity throughout root development seems to relate it specifically to root metabolism and indicates it as a molecular marker of rhizogenesis in M. crystallinum

Pathways of ROS homeostasis regulation in "Mesembryanthemum crystallinum" L. calli exhibiting differences in rhizogenesis

A comparison of the hydrogen peroxide (H2O2) content, proline and betacyanin concentration and activities of some antioxidant enzymes (catalase, superoxide dismutase, guaiacol and ascorbate peroxidases) was made in Mesembryanthemum crystallinum L. calli differing in rhizogenic potential. Callus was induced from hypocotyls of 10-day-old seedlings on a medium containing 1 mg l−1 2,4-dichlorophenoxyacetic acid and 0.2 mg l−1 kinetin, which was either supplemented with 40 mM NaCl (CIM-NaCl medium) or did not contain any salt (CIM medium). The callus obtained on CIM-NaCl was rhizogenic, whereas the callus induced on the medium without salt was non-rhizogenic throughout the culture. The rhizogenic callus differed from the non-rhizogenic callus in lower betacyanin and H2O2 content, but the rhizogenic callus displayed a higher proline level. The activity of H2O2 scavenging enzymes, such as catalase (CAT), ascorbate peroxidase (APX) and guaiacol peroxidase (POD), was markedly higher in the rhizogenic callus than in the non-rhizogenic callus, but the total activity of superoxide dismutase (SOD) was higher in the non-rhizogenic callus than in the rhizogenic callus. Aminotriazole (CAT inhibitor) and diethyldithiocarbamate (SOD inhibitor) were added solely to the CIM and CIM-NaCl media to manipulate the concentration of reactive oxygen species (ROS) in the cultured tissues. Both CAT and SOD inhibitors brought about an increase in H2O2 content in calli cultured on CIM-NaCl and the loss of rhizogenic potential. Conversely, the addition of inhibitors to the medium without salt led to a decrease in H2O2 content. This corresponded with a significant decrease in the endogenous concentration of betacyanins, but did not change the lack of rhizogenic ability

Low-vacuum filtration as an alternative extracellular vesicle concentration method : a comparison with ultracentrifugation and differential centrifugation

Recent years have brought great focus on the development of drug delivery systems based on extracellular vesicles (EVs). Considering the possible applications of EVs as drug carriers, the isolation process is a crucial step. To solve the problems involved in EV isolation, we developed and validated a new EV isolation method—low-vacuum filtration (LVF)—and compared it with two commonly applied procedures—differential centrifugation (DC) and ultracentrifugation (UC). EVs isolated from endothelial cell culture media were characterized by (a) Transmission Electron Microscopy (TEM), (b) Nanoparticle Tracking Analysis (NTA), (c) Western blot and (d) Attenuated Total Reflection Fourier-Transform Infrared Spectroscopy (ATR-FTIR). Additionally, the membrane surface was imaged with Environmental Scanning Electron Microscopy (ESEM). We found that LVF was a reproducible and efficient method for EV isolation from conditioned media. Additionally, we observed a correlation between ATR-FTIR spectra quality and EV and protein concentration. ESEM imaging confirmed that the actual pore diameter was close to the values calculated theoretically. LVF is an easy, fast and inexpensive EV isolation method that allows for the isolation of both ectosomes and exosomes from high-volume sources with good repeatability. We believe that it could be an efficient alternative to commonly applied methods

Metody i obszary zarządzania

Publikacja recenzowana / Peer-reviewed publicationOpracowanie poświęcone jest charakterystyce i propozycji rozwiązania wybranych problemów zarządzania. Kilka tekstów dotyczy roli mediów (a także innych instrumentów) w marketingu. Ciekawe jest opracowanie oparte na pewnej autorskiej typologii instrumentów marketingu-mix stosowych przez przedsiębiorstwa działające na rynkach zagranicznych. Druga grupa artykułów poświęcona jest nowym formom zatrudnienia, partycypacji pracowników oraz standardów ochrony pracy, które winny być stosowane przez nasze organizacje. Zatem blok ten dotyczy zagadnień organizacji pracy. Są także artykuły prezentujące konkretne metody zarządzania, począwszy od zarządzania komunikacją, przez zarządzanie strategiczne, a na systemach zarządzania produkcją skończywszy. Niektórzy Autorzy podejmują się nie tylko prezentacji, a także przedstawiają pewne rozwiązania zagadnień dotyczących niemal wszystkich obywateli. Tu przede wszystkim należy wymienić rozważania o kształtowaniu tran sportu publicznego. Ciekawe jest także studium o możliwości wykorzystania Internetu na rynku ubezpieczeń społecznych

The impact of biotic and abiotic stress factors on development of European ash Tissue cultures

Fraxinus excelsior L. is threatened by a variety of environmental factors causing a decline of the species. The most important biotic factors negatively affecting the condition of the F. excelsior population are fungi such as the pathogen Hymenoscyphus fraxineus. Abiotic factors with potentially harmful effect to the F. excelsior population are the accumulation of heavy metals and salinity in soils. Thus, the aim of this study was to investigate the impact of selected biotic and abiotic stress factors to determine which of them pose a threat to European ash. The study was conducted using in vitro techniques based on callus and seedlings regenerated via indirect organogenesis. Tissue cultures exclude the influence of other factors, including the environmental impact on ash extinction. The results confirmed very strong pathogenic potential of H. fraxineus in which after 14 days the callus tissue cells died as the tissue failed to activate its defense mechanisms. Experiments showed the high toxicity of cadmium in concentration of 0.027 mmol/L. Salinity caused the activity of oxidation enzymes to vary among seedlings and calluses in the control suggesting the enzymes play a role in controlling the morphogenetic development of tissue cultures