Multi-instrumental Investigation of Affecting of Early Somatic Embryos of Spruce by Cadmium(II) and Lead(II) Ions

Abstract

Abstract:The main aim of this work was to use multi-instrumental analytical apparatus to investigate the effects of treatment with cadmium(II) and/or lead(II) ions (50, 250 and 500 µM) for twelve days on early somatic spruce embryos (ESEs). Primarily we used image analysis for estimation of growth and a fluorimetric sensor for enzymatic detection of viability of the treated ESEs. It follows from the obtained results that Cd caused higher toxicity to ESEs than Pb. Besides this fundamental finding, we observed that ESEs grew and developed better in the presence of 500 µM of the metal ions than in the presence of 250 µM. Based on the results obtained using nuclear magnetic resonance this phenomenon was related to an increase of the area of ESE clusters by intensive uptake of water from the cultivation medium, due to dilution of the heavy metal concentration inside the cluster. In addition we studied the glutathione content in treated ESEs by the adsorptive transfer stripping technique coupled with the differential pulse voltammetry Brdicka reaction. GSH contents increased up to 148 ng/mg (clone 2/32) and 158 ng/mg (clone PE 14) after twelve Sensors 2007, 7 744 day long treatment with Cd-EDTA ions. The GSH content was about 150 and 160 % higher in comparison with the ESEs treated with Pb-EDTA ions, respectively. The difference between GSH contents determined in ESEs treated with Pb-EDTA and Cd-EDTA ions correlates with the higher toxicity of cadmium(II) ions.

MRI-Based Visualization of the Relaxation Times of Early Somatic Embryos

“…The embryos constitute a unique plant model system applicable for the study of various types of environmental stresses (including metal ions) under well -controlled experimental conditions [9], [10], [11], [12]. In the investigated ESEs, the researchers have used image analysis to estimate the growth, a fluorimetric sensor to detect the viability, and the MRI technique to facilitate non-destructive measurement of the volume.…”

“…MRI is also employed to monitor water changes in early somatic embryos (ESEs) of spruce [7], [8]. Supalková et al [9] present the results of a twelve-day, multi-instrumental analytical examination of such early somatic embryos treated with cadmium (II) and/or lead (II) ions (50, 250, and 500 microns). In the investigated ESEs, the researchers have used image analysis to estimate the growth, a fluorimetric sensor to detect the viability, and the MRI technique to facilitate nondestructive measurement of the volume.…”

Employment of Electrochemical Techniques for Metallothionein Determination in Tumor Cell Lines and Patients with a Tumor Disease

“…The frozen cells were mixed with extraction buffer (100 mM potassium phosphate, pH 8.7) to a final volume of 1 mL and homogenized using hand-operated homogenizer ULTRA-TUR-RAX T8 (IKA, Germany) placed in an ice bath for 3 min at 25 000 rpm [45]. The homogenate was centrifuged at 10 000 g for 15 min and at 4 8C (Eppendorf 5402, USA).…”

Effects of Reduced Glutathione, Surface Active Agents, and Ionic Strength on the Detection of Metallothioneins by Using of Brdicka Reaction

“…The harvested cells (approximately 100 000 cells) were transferred to a test tube and then deep frozen by liquid nitrogen to disrupt cells. The frozen cells were mixed with extraction buffer (100 mM potassium phosphate, pH 8.7) to a final volume of 1 ml and homogenized using hand-operated homogenizer ULTRA-TURRAX T8 (IKA, Germany) placed in an ice bath for 3 min at 25 000 rpm [32]. The homogenate was centrifuged at 10 000 g for 15 min and at 4 8C (Eppendorf 5402, USA).…”