Callus induction and plant regeneration of Iris dichotoma Pall. in endangered species
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초록
Iris dichotoma Pall. is an important endangered plant belonging to the family Iridaceae. A method was developed for the rapid micropropagation of I. dichotoma through plant regeneration from leaf, rhizome, and root explant-derived calli. Leaf, rhizome, and root segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; 0-3.0 mg･L-1) for callus induction. Callus production was highest at 1.0 mg･L-1 2,4-D, where 73.8% and 45.5% of cultured rhizome and root cuttings, respectively, produced calli. The viable calli were maintained at an induced concentration of 2,4-D (3.0 mg･L-1). They were then transferred to MS medium supplemented with various concentrations of 2,4-D (0-3.0 mg･L-1) in combination with 6-benzyladenine (BA: 0, 1.0 and 3.0 mg･L-1) for adventitious shoot regeneration. The addition of a low concentration of 2,4-D into BA-containing medium significantly increased the frequency of shoot regeneration in leaf, rhizome, and root-derived calli. The highest number of adventitious shoots (26.4 per callus) formed at 0.5 mg･L-1 2,4-D and 1.0 mg/l BA. For rooting of the shoots, half- strength MS medium supplemented with different concentrations of indole 3-butyric acid (IBA) 0-3.0 mg･L-1 was tested. The optimal results were observed using half-strength MS medium supplemented with 1.0 mg･L-1 IBA, on which 98% of the regenerated shoots developed roots with an average of 3.5 roots per shoot within 45 days. The plantlets raised in vitro were acclimatized and transferred to soil with 95% success. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.
Abstract
Iris dichotoma Pall. is an important endangered plant belonging to the family Iridaceae. A method was developed for the rapid micropropagation of I. dichotoma through plant regeneration from leaf, rhizome, and root explant-derived calli. Leaf, rhizome, and root segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; 0-3.0 mg･L-1) for callus induction. Callus production was highest at 1.0 mg･L-1 2,4-D, where 73.8% and 45.5% of cultured rhizome and root cuttings, respectively, produced calli. The viable calli were maintained at an induced concentration of 2,4-D (3.0 mg･L-1). They were then transferred to MS medium supplemented with various concentrations of 2,4-D (0-3.0 mg･L-1) in combination with 6-benzyladenine (BA: 0, 1.0 and 3.0 mg･L-1) for adventitious shoot regeneration. The addition of a low concentration of 2,4-D into BA-containing medium significantly increased the frequency of shoot regeneration in leaf, rhizome, and root-derived calli. The highest number of adventitious shoots (26.4 per callus) formed at 0.5 mg･L-1 2,4-D and 1.0 mg/l BA. For rooting of the shoots, half- strength MS medium supplemented with different concentrations of indole 3-butyric acid (IBA) 0-3.0 mg･L-1 was tested. The optimal results were observed using half-strength MS medium supplemented with 1.0 mg･L-1 IBA, on which 98% of the regenerated shoots developed roots with an average of 3.5 roots per shoot within 45 days. The plantlets raised in vitro were acclimatized and transferred to soil with 95% success. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.
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