Abstract
An efficient protocol providing a dual regeneration pathway via direct shoot organogenesis and somatic embryogenesis for an endangered species, Metabriggsia ovalifolia W. T. Wang, was established from leaf explants. When applied at 2.5 μM, the cytokinins 6-benzyladenine (BA) or thidiazuron (TDZ) and the auxins indole-3-butyric acid (IBA), α-naphthaleneacetic acid (NAA) and indole-3-acetic acid (IAA) could induce shoots when on basal Murashige and Skoog (MS) medium. BA and TDZ could induce more adventitious shoots (19.1 and 31.2/explant, respectively) than NAA (4.6/explant), IBA (5.7/explant) or IAA (6.4/explant). BA and TDZ at 5–10 μM could induce both shoots and somatic embryos. A higher concentration of TDZ (25 μM) induced only somatic embryos (39.8/explant). The same concentration of BA induced both adventitious shoots (23.6/explant) and somatic embryos (9.7/explant). Thus, somatic embryogenesis in this plant needs a high cytokinin concentration (BA; TDZ), as evidenced by histology. Somatic embryos germinated easily when left on the same media, but formed adventitious roots in two weeks on MS supplemented with 0.5 μM NAA, 0.5 μM IBA and 0.1% activated charcoal. Over 93% of plantlets survived following acclimatization and transfer to a mixture of sand and vermiculite (1:1, v/v) in trays.
Highlights
IntroductionThe establishment of an efficient propagation and plant regeneration system is one effective way of circumventing sudden deterioration or loss of a plant’s natural environment, and serves as an ideal tool to preserve and utilize this rare and endangered plant species
Somatic embryogenesis has never been reported for this plant
We used a high concentration of BA and TDZ to successfully induce both shoot organogenesis and somatic embryogenesis
Summary
The establishment of an efficient propagation and plant regeneration system is one effective way of circumventing sudden deterioration or loss of a plant’s natural environment, and serves as an ideal tool to preserve and utilize this rare and endangered plant species. A protocol for shoot organogenesis from leaf explants was reported for M. ovalifolia[5]. To date, no report exists on somatic embryogenesis for this plant. The main focus of this study was to assess the ability of different plant growth regulators (PGRs) to control somatic embryogenesis and shoot organogenesis in M. ovalifolia in a bid to expand the choices for viable in vitro regeneration pathways which would make applied biotechnological applications, such as genetic transformation, feasible
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