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Abstract Mesembryanthemum forsskaolii and M. nodiflorum belong to family Aizoaceae and were collected from Petrified Forest Protected Area, Egypt. The present study was carried out on the two species to study the genetic variation of four Mesembryanthemum accessions and studying biological activities of their water and ethanolic extracts. The obtained results were summarized as follow: Molecular study Four accessions of Mesembryanthemum were investigated using two molecular markers RAPD and ISSR. RAPD analysis: A Total of 72 reproducible bands were obtained from 8 random decamer primers with different Mesembryanthemum accessions. All of them were polymorphic. Percentage of total polymorphism was found to be 100%. The average number of bands per primer was 9. Primer OPC-06 recorded the highest PIC value (0.43) and primer OPO-12 revealed the lowest PIC value (0.37). The maximum RP was 9.5 which observed with primer OPA- 02 and OPO-09 and the lowest RP value 3 was produced by primer OPO-12. Cluster analysis was performed based on genetic distance by applying unweighted pair group method with arithmetic average (UPGAMA) using two softwares NTSYS and Past 3. The dendrogram obtained from NTSYS split the accessions in two clusters. The first cluster consisted of M. forsskaolii with yellow color and M. nodiflorum. The second cluster included M. forsskaolii with green color and M. forsskaolii with red color. The Past3 software generated two clusters. The first cluster included two groups, the first group contained M. forsskaolii (red) and the second group enclosed M. forsskaolii (green) and M. nodiflorum. The second cluster included M. forsskaolii (yellow). ISSR analysis: A total of 73 marker fragments were generated from 12 ISSR primers. 70 bands were polymorphic with an average 5.38 product per primer and 3 bands were monomorphic. Primer 844B produced the maximum PIC value (0.47) and primer HB15 generated the least PIC value (0.32). The resolving power (RP) value ranged from 1 to 9 generated by primers 814 and HB8 respectively. The dendrogram obtained from NTSYS and Past 3 softwares clustered the accessions in two clusters. The first cluster consisted of M. forsskaolii (yellow) and M. nodiflorum while the second cluster included M. forsskaolii (green) and M. forsskaolii (red). Potential of plant regeneration: Several explants were used in this study such as fresh seeds, dry seeds, buds and leaves. Fresh seeds gave the highest shoot length 3.45 and 4.05 of plantlets of M. forsskaolii and M. nodiflorum respectively. Leaves failed to produce shoot in both species. The different concentrations of 2,4-D and NAA were used to induce callus from explants. Leaves explants only were able to induce callus. Medium contained 1mg/l 2,4-D and 2mg/l NAA) gave the best response of callus formation.Phytochemical screening: The present analysis performed the extraction of M. forsskaolii and M. nodiflorum (wild and callus) using ethanol and hot distilled water. This study has explored various phytochemical compositions; the results showed that the wild plant and callus extracts contained similar compounds. M. forsskaolii extracts contained flavonoids, unsaturated sterols, steroids and alkaloids. M. nodiflorum contained unsaturated sterols, triterpenes, steroids, alkaloids and glycosides. Potential of biological activities: Cytological studies (in vivo): There was a significant depression in the mitotic index percentage (MI) of Allium cepa L. root tips after treatment with water extract of wild and callus from M. forsskaolii and M. nodiflorum. Prophase stage recorded the highest percentage after treatment with water extracts of wild and callus from M. forsskaolii and M. nodiflorum. The increase of the percentage of total abnormalities was significant after all treatments. There were three types of chromosomal aberrations; common type of abnormalities observed during the study was stickiness while, the other types were irregular prophase and spindle disturbance. Cytotoxic activity (in vitro): The cytotoxic activity of the water and ethanol extracts of wild and callus from M. forsskaolii and M. nodiflorum against HELA and MCF-7 cell lines was assessed by MTT assay. The results proved that the growth of the cell lines was inhibited in dose dependent. The ethanol extract showed a moderate cytotoxic potential activity against the two cell lines. The wild plants were more active than callus. |