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Effects of light, sucrose, and cytokinins on somatic embryogenesis in Lilium ledebourii (Baker) Bioss. via transverse thin cell-layer cultures of bulblet microscales

Summary This is the first report describing the culture conditions necessary to induce somatic embryogenesis and plantlet regeneration from transverse thin cell-layers (tTCL) of the rare and endangered bulb species, Lilium ledebourii (Baker) Boiss. (Liliaceae). The tTCLs were transferred onto 1.0 Murashige and Skoog medium (MS) containing various sucrose concentrations [3.0, 4.5, or 6.0% (w/v)] and different combinations of two cytokinins [6-benzylaminopurine (BA) or thidiazuron (TDZ)] with 1.0 µM -naphthaleneacetic acid (NAA) in the dark, or exposed to light (40 µmol m–2 s–1). The aims of this work were to provide an improved propagation method torescue L. ledebourii, and to determine the effects of sucrose concentration, light, and different cytokinins on somatic embryogenesis. Embryogenic callus cultures were obtained only when the tTCLs were transferred onto 1.0 MS medium containing 1.0 µM NAA, various levels of BA (0.4, 1.1, or 2.2 µM), and sucrose [3.0, 4.5, or 6.0% (w/v)] after 3 months culture in the light or in darkness. Combinations of various concentrations of TDZ and NAA did not generate embryogenic calli. The highest rate of growth of embryogenic calli was achieved on 1.0 MS medium supplemented with 1.0µM NAA, 1.1 µM BA, and 4.5% (w/v) sucrose, in the light. Embryo-like structures were grown into plantlets after transfer onto 1.0 MS medium without any plant growth regulators and incubated in the light. Regenerated plants were successfully acclimatised to ex vitro conditions, with a survival rate of 90%.