Abstract
Anthurium andraeanum Lind. is the second most important tropical flower in the world flower market. Somatic embryogenesis and plant regeneration in Anthurium has been reported previously; however, a stable and effective method for its commercial use has not been available. In this study, an efficient somatic embryogenesis and liquid culture system for large-scale production of A. andraeanum seedlings was achieved. Building on previous research, this study investigated the main factors for proembryogenic mass (PEM) proliferation, somatic embryo (SE) development, and SE germination in Anthurium. The results showed that relatively low concentrations of plant growth regulators, mineral nutrition, and sucrose promoted PEM proliferation, SE formation, and germination in a liquid culture system. This system can be described as follows: PEMs were induced from leaf blade explants on Murashige & Skoog (MS) medium with half-strength MS macronutrients (1/2 MS) containing 2.0 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.5 mg L-1 kinetin (KT), and 3% sucrose and were proliferated in ½ MS liquid medium containing 1.0 mg L-1 2,4-D, 0.5 mg L-1 KT, and 3% sucrose. The highest proliferation coefficients were 5.11–5.16. PEMs were then transferred to MS medium with 1/8 MS macronutrients (1/8 MS) liquid medium containing 1% sucrose to develop into globular embryos and mature embryos. Finally, the mature embryos were placed on four layers of absorbent filter paper saturated with 1/8 MS liquid medium containing 1% sucrose for germination, and an average of 60 seedlings per gram SEs was obtained. This liquid culture system can be used in large-scale and synchronic production of Anthurium seedlings.
Highlights
Anthurium andraeanum Lind. belongs to the Araceae family
We concluded that 1/2 Murashige & Skoog (MS) supplemented with 1.0 mg L−1 2,4-D, 3% sucrose, and 0.5 mg L−1 KT was the optimal medium for proembryogenic mass (PEM) proliferation in liquid culture
Somatic embryogenesis and plant regeneration in solid culture has been reported in Anthurium (Kuehnle et al, 1992; Xin et al, 2006; Bautista et al, 2008; Pinheiro et al, 2013, 2014; Bhattacharya et al, 2016), but a complete liquid culture system including proliferation of logarithmic growth was observed at the early stage of proliferation (Figure 2A)
Summary
Anthurium andraeanum Lind. belongs to the Araceae family. It is famous for its showy spathe and beautiful foliage and has been the second most important tropical flower (next to orchid) in the world flower market (Dufour and Guerin, 2003; Teixeira da Silva et al, 2015). Anthurium tissue cultures have been established by organogenesis from leaf, petiole, spathe, or axillary bud explants, and multiple shoots can be proliferated through a repeated callus-induction and regeneration process or axillary shoot proliferation (Pierik, 1976; Pierik et al, 1979; Geier, 1990; Martin et al, 2003; Puchooa, 2005; Teixeira da Silva et al, 2015). This method has several disadvantages, including low propagation efficiency, occasional somaclonal variation, and inhomogeneity (Kuehnle et al, 1992). Somatic embryogenesis is an effective micropropagation method that can overcome the shortcomings described above and has been successfully used for many kinds of economically valuable plants (Merkle et al, 1990; Duoue et al, 2006; Klimaszewska et al, 2015)
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