A method for rapid micropropagation of Ophiopogon japonicus through plant regeneration from leaf, rhizome and root explants derived calli has been developed. Leaf, rhizome and root segments were cultured on Murashige and Skoog (MS) medium supplemented with plant growth regulators (2,4-D, NAA, IAA; 0~3.0 mg/ L) for callus induction. Callus production was highest at 1.0 mg/L 2,4-D where 91%. These calli were transferred to MS medium supplemented with various concentrations of 2,4-D (0, 0.5, 1.0, 3.0 mg/L) in combination with 6-benzyladenine (BA: 0, 0.5, 1.0 and 3.0 mg/L) for adventitious shoot regeneration. The addition of low concentration of 2,4-D into BA containing medium significantly increased the frequency of shoot regeneration in both leaf, rhizome and root derived calli. The highest frequency of adventitious shoot (88%) formation was on MS medium supplemented with 0.5 mg/L 2,4-D and 1.0 mg/L BA. For rooting of the shoots, half-strength MS medium supplemented with different concentrations of plant growth regulators (2,4-D, IBA, NAA) 0, 0.1, 1.0, 3.0 mg/L was tried. The optimal results was observed on half-strength MS medium supplemented with 3.0 mg/L NAA (average of 42.9 roots per explant). In vitro raised plantlets were acclimatized and transferred to soil with 100% success. This in vitro propagation protocol would be useful for conservation as well as mass propagation of this medical plant.
This experiments were carried out to find out the effects of different explant materials, kinds and concentration of plant growth regulators on the in vitro regeneration of Iris pseudoacorus L. The effects of growth regulators on regeneration from three explant sources (root, rhizome and leaf) were more or less same. Explants produced only callus with 2,4-D treatment and other regulators had no effects. The highest frequency of callus induction (rhizome: 84%) was on MS media supplemented with 1.0 mg/L 2,4-D. For plant regeneration, when calli were transferred to the first media supplemented with 1.0 mg/L BA and 1.0 mg/L 2,4- D. Regenerated plants were successfully acclimatized in soil containing a mixture of vermiculite : perlite : leafmold (1 : 1 : 1), and their survival was high.
When the leaves, roots and stem segments of seedling of Polygonatum odoratum were cultured on Murashige and Skoog medium with 2.0mg/l BAP, stem segments were the most efficient explants for adventitious shoot inductino. To observe the efficient combination of growth regulators on the adventitious shoot formation , stem segments were cultured on MS medium with various kinds of cytokinins (BAP, kinetin, zeatin). From this experiment, cytokinin treatement was prerequisite for theadventitious shoot formatino,especially BAP was the most effective. Auxin (NAA or IBA) in combination with cyotokinin highly enhanced the adventitious shoot formation. Twenty five percents of explants produced the adventitious shoots on medium with 2.0mg/l BAP solely, while 83% of explants produced the adventitious shoots on medium with 2.0mg/l BAP and 0.1mg/l IBA. Root formationform adventitious shoot was promoted after transfer to 1/2 MS medium supplemented with 0.1mg/l IBA and 0.5mg/l zeatin, thereafter the plantlets with shoots and roots were cultured on 1/2MS medium lacking growth regulators. When the stem segments were cultured to MS medium with 1.0mg/l 2,4 NAA and IBA , yellow and nodulous cali were formed from the stem segments which were developed into adventitious roots. These roots were actively grew after transferred to MS liquid medium lacking growth regulators.