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Callus Induction of Ocimum sanctum and Estimation of Its Total Flavonoids Content

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Author(s): Dr. Anna Pick Kiong Ling

Journal: Asian Journal of Agricultural Sciences
ISSN 2041-3882

Volume: 1;
Issue: 2;
Start page: 55;
Date: 2009;
Original page

Keywords: Anti-oxidant | essential oil | Holy Basil | Ocimum sanctum | orientin | vicenin

ABSTRACT
Ocimum sanctum, also known as “H oly Basil” is one of the most common medicinal plants used bydiverse cultures and tribal groups. Callus induction from leaf explants of O. sanctum was conducted byincubating leaf explants on Murashige and Skoog (MS) medium supplemented with 2, 4!dichlorophenoxyaceticacid (2,4!D), picloram, and indole-butyric acid (IBA) at 0, 1, 3, and 5 mg/L as well as the combination of 3mg/L picloram with different concentrations (0, 0.5, 1.0, 1.5, and 2.0 m g/L) of 6!benzylaminopurine (BAP)or kinetin. Results obtained from the studies revealed that all the leaf explants incubated on phytohormonesupplementedmedium formed callus. Leaf explants grown on 3 mg/L picloram formed callus after 8±1 daysof culture, and degree of callus formation w as found to be the highest (++++) among all the single auxintreatments. In contrast, the degree of callus formed from the leaf explants cultured on MS mediumsupplemented with combination of auxin and cytokinins were evidently lower than those in the single auxintreatments. Leaf explants cultured on kinetin-supplemented MS medium showed a higher degree of callusformation (++++) as compared to BAP-supplemented MS medium. There was no significant difference betweenthe days of callus formation among all the cytokinin-supplemented treatments. The total flavonoids content ofleaf-derived callus cultured on 3 mg/L picloram w ere also estimated and compared with the in vivo leaf tissues.The aluminium (III) chloride colorimetric assay revealed that the total flavonoids content of in vivo leaf tissuesof O. sanctum were 2.2 times higher than leaf-derived callus, whereby the former produced 0.733±0.077miligram catechin equivalent per gram fresh weight (mg CE/g fresh weight), while the latter only yielded0.333±0.043 mg CE/g fresh weight.

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