Molecular Analysis of Ocimum sanctum Induce Antigenotoxicity on Allium cepa Grown in Slurry Water

Abstract

Use of industrial effluents and sewage sludge for agriculture have become a common practice in world and India too as a result of which many harmful metals and substances get transferred and accumulated into plant tissues from soil and causing many problems and diseases in living organisms.Management of these abnormalities/ diseases with medicinal plant is an ancient practice that has gained momentum in recent years. Aqueous extract of Ocimum sanctum (Tulsi) have long been used for traditional management of cancer due to its antigenotoxic nature. Data of genotoxicity and carcinogenicity of are rather controversial, depending on the genetic system or the assay used. Therefore, in present study, the genotoxicity profile of five different concentration of slurry water (20%, 40%, 60%, 80% and 100%) and antigenotoxic profile of three concentration of aqueous extract of tulsi leaves were evaluated with two genotoxicity assays. First assay by using root tip cells and leaves of Allium cepa plant. The parameters evaluated in cytological assay were number of cells in dividing stage, mitotic indices (MI), % mitodepressive (MD) effect, RDR and chromosomal aberrations (CA) on A. cepa roots. The results demonstrated both cyto as well as genotoxicity towards A. cepa meristematic cells, inducing chromosomal changes. Second assay i.e. molecular assay was performed with 20primer pairs of cytochrome P450 based functional molecular using leaves of onion, extracted after 30 days, 45 days and 60 days of the exposure to different concentration of both the test chemicals. The mutagenic activity of the active compounds was demonstrated by the detection of DNA changes electrophorogram of the treated cells compared to the control. The PCR profiles showed a great variation in banding patterns, particularly in intensity, number, thickness and mobility of the generated DNA fragments by the used primers. GTS value served as a good indicator of genotoxic effect of slurry water as well as antigenotoxic effect of tulsi extract. Together, these results contribute to a better understanding of the damage caused by these substances in living organisms and reveal a promising strategy for prospective studies of the toxic and anti toxic effect of the selected samples and plant extracts. Keywords :Slurry water, industrial effluent, Mitotic Index (MI), mitodepressive (MD) effect, chromosomal aberrations (CA), A. cepa, Ocimum sanctum, Mutagenicity, Cytochrome P450 marker, functional molecular marker, Genomic Template Stability (GTS), antigenotoxicity.