Direct and indirect in-vitro plant regeneration and antioxidant production in tsaang gubat(Carmona retusa (Vahl.) Masam)
2010
Reglos-Zara, R.A. | Cedo, M.L.O. | de Guzman, C.C. | Rodriguez, F. M., Philippines Univ. Los Baños, College, Laguna (Philippines). Crop Physiology Division
In vitro plant regeneration of tsaang gubat(Carmona retusa (Vahl.) masam) via direct and indirect methods of using shoot tip explants was studied. The level of antioxidants such as tannin, flavonoids and phenolics in the leaves of in-vitro and field-grown plants was also evaluated. Multiple shoot formation was enhanced when shoot tips were cultured in Murashige and Skoog's (MS) medium containing 5 mg/L 6-benzylaminopurine (BAP). Higher BAP concentration did not induced more new shoots; instead, the original shoot inoculum appeared stunted, vitrified and had abnormally large leaves. Continuous subculturing (6-8 wk) of the regenerated shoots to MS + 5 mg/L BAP and 15%(V/V) coconut water induced shoot proliferation with an average of 15-20 shoots per inoculum after the 3rd subculture. MS medium containing 0.5 mg/L napthalene acetic acid(NAA) and 4 mg/L BAP was found optimum for callus formation. Subsequent differentiation of shoots was observed only in media containing BAP with the highest number of shoots produced at 10 mg/L. Regenerated shoots were successfully rooted on indole-3-butyric acid(IBA)-supplemented medium; the longest and highest number of roots was recorded at 10 mg/L IBA. Levels of phenolics, flavonoids and tannins were higher in the leaves of tissue-culture-raised plants than those of in vivo grown plants. Likewise, plants maintained for long-term (24 wk) under in vitro conditions produced these secondary metabolites, although at lower levels compared with the field-grown materials.
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