A wide range of genotoxicity assays are available to determine the DNA damage in plant cells caused by allelochemicals. However most of them, sometimes present some technical difficulties in interpretation of results. Acridine orange/ethidium bromide double staining assay is proposed as a rapid, inexpensive and easy-to-perform assay to investigate the apoptotic damage from phenols in plant cells. Our data support the validity of this assay and the hypothesis that phenols induce unrepairable severe genetic alterations due to apoptosis. The proapoptotic effects of Olive oil Mill wastewaters (OMWW, rich in polyphenols and catechol) were investigated on radish root cells. In 25% OMWW treated root cells, strong DNA fragmentations and hypersegmented nuclei were observed and increase in percentage of total apoptotic cells was registered during 24 h. The root cells treated with 12.5% OMWW showed the typical apoptotic hallmarks, but the percentage of total apoptotic cells decreased from 92% to 60% during 24 h, due to the significant decrease in early apoptotic cells and a concomitant increase in cell viability. The catechol at 10-4 to 10-3 M concentrations caused dose-dependent effect, nuclear damage increased during the 24 h and the root cells showed drastic chromatin disintegrations.
A wide range of genotoxicity assays are available to determine the DNA damage in plant cells caused by allelochemicals. However most of them, sometimes present some technical difficulties in interpretation of results. Acridine orange/ethidium bromide double staining assay is proposed as a rapid, inexpensive and easy-to-perform assay to investigate the apoptotic damage from phenols in plant cells. Our data support the validity of this assay and the hypothesis that phenols induce unrepairable severe genetic alterations due to apoptosis. The proapoptotic effects of Olive oil Mill wastewaters (OMWW, rich in polyphenols and catechol) were investigated on radish root cells. In 25% OMWW treated root cells, strong DNA fragmentations and hypersegmented nuclei were observed and increase in percentage of total apoptotic cells was registered during 24 h. The root cells treated with 12.5% OMWW showed the typical apoptotic hallmarks, but the percentage of total apoptotic cells decreased from 92% to 60% during 24 h, due to the significant decrease in early apoptotic cells and a concomitant increase in cell viability. The catechol at 10-4 to 10-3 M concentrations caused dose-dependent effect, nuclear damage increased during the 24 h and the root cells showed drastic chromatin disintegrations.
Acridine orange/Ethidium bromide double staining test: A simple In-vitro assay to detect apoptosis induced by phenolic compounds in plant cells
Ciniglia C;
2010
Abstract
A wide range of genotoxicity assays are available to determine the DNA damage in plant cells caused by allelochemicals. However most of them, sometimes present some technical difficulties in interpretation of results. Acridine orange/ethidium bromide double staining assay is proposed as a rapid, inexpensive and easy-to-perform assay to investigate the apoptotic damage from phenols in plant cells. Our data support the validity of this assay and the hypothesis that phenols induce unrepairable severe genetic alterations due to apoptosis. The proapoptotic effects of Olive oil Mill wastewaters (OMWW, rich in polyphenols and catechol) were investigated on radish root cells. In 25% OMWW treated root cells, strong DNA fragmentations and hypersegmented nuclei were observed and increase in percentage of total apoptotic cells was registered during 24 h. The root cells treated with 12.5% OMWW showed the typical apoptotic hallmarks, but the percentage of total apoptotic cells decreased from 92% to 60% during 24 h, due to the significant decrease in early apoptotic cells and a concomitant increase in cell viability. The catechol at 10-4 to 10-3 M concentrations caused dose-dependent effect, nuclear damage increased during the 24 h and the root cells showed drastic chromatin disintegrations.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
