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This paper presents the results of a study on the influence of lead (Pb2+) on DNA integrity on plant cells. The study was performed on the root tips of lupin (Lupinus luteus cv. Juno) seedlings treated with two selected concentrations of Pb(NO3)2: 150 and 350 mg l-1, which were found to inhibit root growth by 50% and 70%, respectively [Ruciłska et al. Plant Physiol. Biochem. 37 (1999) 37187-37194]. Roots exposed to those external lead concentrations took up about 50 and 70 mg l-1 Pb2+ g-1 fresh weight (FW) over 48 h of incubation. A dose-dependent increase in the degree of root injury was observed in the presence of both tested concentrations. The genotoxicity of lead in lupin root cells was analysed using a mild alkaline comet assay at pH 12.3, which allows the detection of single strand breaks. The quantity of the DNA fragments migrating away from the nuclear remnant (tail area) increased proportionally to the lead content inside the roots, and was positively correlated with the degree of root injury. At 150 mg l-1 Pb2+, a high frequency distribution of nuclei having large values of tail lengths and moments was observed. By contrast, the number of nuclei with minimum values of these parameters increased at 350 mg l-1 Pb2+. This data suggests that lead at low concentrations induces the formation of short, rapidly migrating DNA fragments, whereas at higher concentrations, lead probably causes other changes to DNA that result in slower DNA migration in the electric field.
Root meristem nucleoli from soybean (Glycine max. cv. Aldana) seedlings germinated for 3 days at 25°C and then for 4 days at 10°C or still at 25°C (control) were examined. Chill was observed to reduce root meristematic zone growth 15-fold. Nucleoli doubled their volume at 10°C. Autoradiographic studies showed that after 20 min 3H-uridine incubation at 10°C, incorporation of this precursor (postincubation time 0) into nucleoli of chilled seedlings was 4.7 times weaker than in the control. After 80 min postincubation in nonradioactive medium, the cytoplasm became the most intensely labelled cell area in the control material, while in chilled roots the nucleoli were still most intensely labelled and the cytoplasm was 11 times less labelled than in the control. The increase in nucleoli volume at 10°C is suggested to result from greater cold-induced inhibition of the dynamics of maturation and transport of ribosome subunits than of rRNA synthesis dynamics. Ultrastructural studies of chilled seedling nucleoli showed a significant decrease in the fibrillar component and an increase in the granular component, forming characteristic clusters. They are supposed to correspond to shortened and condensed pre-rRNA transcription complexes (compacted "Christmas trees").
Aluminium toxicity is one of the major factors that limit plant growth and develop­ment in many acid soils. Root cells plasma membrane, particularly of the root apex, seems to be a major target of A1 toxicity. However, strong interaction of A13+ , the main A1 toxic form, with oxygen donor ligands (proteins, nucleic acids, polysaccharides) re­sults in the inhibition of cell division, cell extension, and transport. Although the iden­tification of A1 tolerance genes is under way, the mechanism of their expression re­mains obscure.
Vascular storage parenchyma cells of carrot roots were treated with methanol solutions of radicinin and epiradicinol produced by Alternaria radicina and with alternariol and alternariol methyl ether produced by A. alternata at concentrations of 25 µg/ml and 250 µg/ml, as well as culture filtrates of both fungi. Cell ultrastructure was observed by TEM. No visible changes were noted after treatment with 25 µg/ml toxin solutions. The most extensive plication of cell membranes, and sometimes also cell walls, and the formation of numerous vesicles in the cytoplasm, was observed in cells treated with the higher concentration of toxins. Plasma membrane withdrawal and vésiculation, microvacuole formation, and accumulation of plastoglobuli in chromoplasts also occurred. No changes in the structure of endoplasmic reticulum and dictyosomes were noted. The responses of cell structures to particular toxins were nonspecific. Treatment with culture filtrates from A. radicina resulted in the occurrence of osmiophilic, electron-dense substance in the cytoplasm and plastoglobuli. All alterations induced by filtrates were more extensive than those resulting from toxin solutions, but membrane integrity was not disturbed after any of the treatments.
Lead, similar to other heavy metals and abiotic factors, causes many unfavorable changes at the subcellular and molecular levels in plant cells. An increased level of superoxide anion in Pisum sativum root cells treated with 1 mM Pb(NO3)2 evidenced oxidative stress conditions. We found increased activities of enzymatic components of the antioxidative system (catalase and superoxide dismutase) in the cytosol, mito- chondrial and peroxisomal fractions isolated from root cells of Pisum sativum grown in modified Hoagland medium in the presence of lead ions (0.5 or 1 mM). Two isoenzyme forms of superoxide dismutase (Cu,Zn-SOD and Mn-SOD) found in differ­ent subcellular compartments of pea roots were more active in Pb-treated plants than in control. Increased amount of alternative oxidase accompanied by an increased ac­tivity of this enzyme was found in mitochondria isolated from lead-treated roots. These results show that plants storing excessive amounts of lead in roots defend them­selves against the harmful oxidative stress caused by this heavy metal.
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