Abstract
Mature zygotic embryos of Cycas revoluta Thunb. were used as explants to investigate direct and indirect organogenesis. Explants were incubated on half-strength Murashige and Skoog (½ MS) basal medium supplemented with various plant growth regulators, singly or in combination (all at 0.5 mg l-1): 6-benzyladenine (BA), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), Kin×2,4-D, BA×Kin and BA×2,4-D. Cultures were placed at a low light intensity (4 µmol m-2 s-1 PPFD). Adventitious shoot regeneration was observed in the presence of 0.5 mg l-1BA after 35 days. The highest number of direct and indirect shoots per zygotic embryo was 3.67 and 29.67, respectively. Roots were induced on indirect shoots by continuous culture on rooting medium (½ MS, 0.1 mg l-1 1-naphthaleneacetic acid) and hardened successfully in perlite. Each rooted plantlet with pinnate leaves and a primary tap root was individually isolated and acclimatized 185 days after the beginning of culture, with a 10% success rate.
Highlights
Cycads, an ancient group of plants that has survived from the Permian era and flourished in the Mesozoic and Jurassic periods, are often referred to as ‘living fossils’ (Gilbert, 1984)
The basal culture medium was Murashige and Skoog (MS; Murashige and Skoog, 1962) macro- and microelements at half the concentration (i.e., 1⁄2 MS), MS vitamins, 30 g l-1 sucrose and solidified with 0.6% (w/v) agar. 1⁄2MS was supplemented with several plant growth regulators, alone and in combination, all at 0.5 mg l-1: BA, Kin, 2,4-D, Kin × 2,4-D, BA × Kin and BA × 2,4-D
The objective of this experiment was to establish a simplified protocol for shoot production from C. revoluta zygotic embryos (ZEs)
Summary
An ancient group of plants that has survived from the Permian era and flourished in the Mesozoic and Jurassic periods, are often referred to as ‘living fossils’ (Gilbert, 1984). Of the Cycadaceae family, commonly known as "Japanese sago palm", is native to southern China and Japan (Stevenson, 1990). Cycads are dioecious with males bearing cones and females bearing groups of megasporophylls (Ikeno, 1896). Plants are propagated either from seeds, which remain viable for only a short time, or from vegetative offshoots. C. revoluta demonstrates rapid loss of seed viability and low morphogenic potential, which hinder its conservation as well as favour an effective and rapid mass propagation. In vitro plant regeneration could be a way to conserve this plant and other endangered species (Ling et al, 2008)
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