An efficient plant propagation system through nodal explants was established in Ocimum gratissimum L, a medicinally important herbaceous perennial herb belonging to the family Lamiaceae. Axillary shoot bud proliferati...An efficient plant propagation system through nodal explants was established in Ocimum gratissimum L, a medicinally important herbaceous perennial herb belonging to the family Lamiaceae. Axillary shoot bud proliferation was initiated from nodal explants cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of N6-benzyladenine (BA) (0.5 - 3.0 mg/l), Kinetin (KN) (0.5 - 3.0 mg/l) and 2-isoPentenyladenine (2-iP) (0.5 - 3.0 mg/l). Maximum numbers of shoots (5.17 ± 0.04) with average length (2.50 ± 0.07) were induced on medium containing 1.0 mg/l BA. Shoot multiplication was maintained by repeated subculturing the original nodal explants on shoot multiplication medium after each harvest of newly formed shoots. Histological study shows that the organogenesis occurs directly, without callus formation on epidermal and sub epidermal layer of the explants. Rooting of shoots was achieved on half strength MS medium supplemented with 1.5 mg/1 Indole-3-butyric acid (IBA) and 2% sucrose. Well-developed complete plantlets were transferred to plastic pots containing a mixture of (1:1) soil and vermiculite showed 82.5 % survival rate. Genetic fidelity was assessed by chromosome analysis and DNA fingerprinting using random amplified polymorphic DNA (RAPD) of in vitro and in vivo plants. Nine arbitrary decamers displayed same banding profile showed no genomic alterations, indicating homogeneity among the tissue culture regenerates and genetic uniformity with that of donor plants. The present study provides high fidelity micro-propagated system for efficient and rapid micro-propagation protocol of this important medicinal plant and great use in conserving without risk of genetic instability.展开更多
A rapid and improved micropropagation protocol was developed for Curcuma sp., a threatened and high value medicinal plant, using main bud sprout and top of rhizome sprout as explants. Stepwise optimization of differen...A rapid and improved micropropagation protocol was developed for Curcuma sp., a threatened and high value medicinal plant, using main bud sprout and top of rhizome sprout as explants. Stepwise optimization of different plant is to change the growth regulator, reduce the level of macro-nutrition and add humate. The present study has created multiple shoot and root induction and plantlet in vitro culture, transfers the plantlet to ex vitro. The M2 medium (MS’s macronutrition 1/4, MS’s micronutrition + Morel’s vitamine + coconut milk 10% + sucrose 25 g/l + humate 1.0 ml/l + agar 7.5 g/l + 2,4D 0.5 mg/l + BAP1.0 mg/l + TDZ 1.0 mg/l) is the highest ratio of callus induction. The TA3 medium (MS’s macronutrition 1/4 + MS’s micronutrition + Morel’s vitamin+ coconut milk 10% + sucrose 25g/l + humate 2.0 ml/l + 2,4 D 0.5 mg/l + Kinetin 2.0 mg/l + TDZ 1.0 mg/l + BAP 1.0 mg/l2 + NAA 2.0 mg/l + activated carbon 2.0 g/l) is able to create buds and regeneration multiple bud for Curcuma sp. TA3 medium adding IAA 2 mg/l and IBA 0.5 mg/l has resulted in the highest indices of quantity, healthy shoot and large diameter of roots. A large number of healthy plantlets are induced by the medium of MS’s macronutrients 1/2, MS’s micronutrients full, Morel’s vitamin, humate 3 g/l, coconut milk 150 ml/l, activated carbon 3 g/l, composition phytohormone: IAA 2 mg/l + BAP 2 mg/l + TDZ 0.5 mg/l. Further studies should focus on optimizing the humate concentration for these species of Zingiberaceae. The duration of the research is from 5/2015 to 4/2016 at Faculty of Agriculture and Forestry, Tay Nguyen University, Vietnam.展开更多
文摘An efficient plant propagation system through nodal explants was established in Ocimum gratissimum L, a medicinally important herbaceous perennial herb belonging to the family Lamiaceae. Axillary shoot bud proliferation was initiated from nodal explants cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of N6-benzyladenine (BA) (0.5 - 3.0 mg/l), Kinetin (KN) (0.5 - 3.0 mg/l) and 2-isoPentenyladenine (2-iP) (0.5 - 3.0 mg/l). Maximum numbers of shoots (5.17 ± 0.04) with average length (2.50 ± 0.07) were induced on medium containing 1.0 mg/l BA. Shoot multiplication was maintained by repeated subculturing the original nodal explants on shoot multiplication medium after each harvest of newly formed shoots. Histological study shows that the organogenesis occurs directly, without callus formation on epidermal and sub epidermal layer of the explants. Rooting of shoots was achieved on half strength MS medium supplemented with 1.5 mg/1 Indole-3-butyric acid (IBA) and 2% sucrose. Well-developed complete plantlets were transferred to plastic pots containing a mixture of (1:1) soil and vermiculite showed 82.5 % survival rate. Genetic fidelity was assessed by chromosome analysis and DNA fingerprinting using random amplified polymorphic DNA (RAPD) of in vitro and in vivo plants. Nine arbitrary decamers displayed same banding profile showed no genomic alterations, indicating homogeneity among the tissue culture regenerates and genetic uniformity with that of donor plants. The present study provides high fidelity micro-propagated system for efficient and rapid micro-propagation protocol of this important medicinal plant and great use in conserving without risk of genetic instability.
文摘A rapid and improved micropropagation protocol was developed for Curcuma sp., a threatened and high value medicinal plant, using main bud sprout and top of rhizome sprout as explants. Stepwise optimization of different plant is to change the growth regulator, reduce the level of macro-nutrition and add humate. The present study has created multiple shoot and root induction and plantlet in vitro culture, transfers the plantlet to ex vitro. The M2 medium (MS’s macronutrition 1/4, MS’s micronutrition + Morel’s vitamine + coconut milk 10% + sucrose 25 g/l + humate 1.0 ml/l + agar 7.5 g/l + 2,4D 0.5 mg/l + BAP1.0 mg/l + TDZ 1.0 mg/l) is the highest ratio of callus induction. The TA3 medium (MS’s macronutrition 1/4 + MS’s micronutrition + Morel’s vitamin+ coconut milk 10% + sucrose 25g/l + humate 2.0 ml/l + 2,4 D 0.5 mg/l + Kinetin 2.0 mg/l + TDZ 1.0 mg/l + BAP 1.0 mg/l2 + NAA 2.0 mg/l + activated carbon 2.0 g/l) is able to create buds and regeneration multiple bud for Curcuma sp. TA3 medium adding IAA 2 mg/l and IBA 0.5 mg/l has resulted in the highest indices of quantity, healthy shoot and large diameter of roots. A large number of healthy plantlets are induced by the medium of MS’s macronutrients 1/2, MS’s micronutrients full, Morel’s vitamin, humate 3 g/l, coconut milk 150 ml/l, activated carbon 3 g/l, composition phytohormone: IAA 2 mg/l + BAP 2 mg/l + TDZ 0.5 mg/l. Further studies should focus on optimizing the humate concentration for these species of Zingiberaceae. The duration of the research is from 5/2015 to 4/2016 at Faculty of Agriculture and Forestry, Tay Nguyen University, Vietnam.
