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Regeneration of Aeschynanthus radicans via direct somatic embryogenesis and analysis of regenerants with flow cytometry

In Vitro Cellular & Developmental Biology - Plant volume 45, pages 34–43 (2009)Cite this article

Abstract

Plant regeneration through direct somatic embryogenesis in Aeschynanthus radicans ‘Mona Lisa’ was achieved in this study. Globular somatic embryos were formed directly from cut edges of leaf explants and cut ends or on the surface of stem explants 4 wk after culture on Murashige and Skoog (MS) medium supplemented with N-phenyl-N′-1, 2, 3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA), TDZ with 2,4-dichlorophenoxyacetic acid (2,4-D), or 6-benzylaminopurine (BA) or kintin (KN) with 2,4-D. MS medium containing 9.08 μM TDZ and 2.68 μM 2,4-D resulted in 71% of stem explants producing somatic embryos. In contrast, 40% of leaf explants produced somatic embryos when induced in medium containing 6.81 μM TDZ and 2.68 μM 2,4-D. Somatic embryos matured, and some germinated into small plants on the initial induction medium. Up to 64% of stem explants cultured on medium supplemented with 9.08 μM TDZ + 2.68 μM 2,4-D, 36% of leaf explants cultured on medium containing 6.81 μM TDZ and 2.68 μM 2,4-D had somatic embryo germination before or after transferring onto MS medium containing 8.88 μM BA and 1.07 μM NAA. Shoots elongated better and roots developed well on MS medium without growth regulators. Approximately 30–50 plantlets were regenerated from each stem or leaf explant. The regenerated plants grew vigorously after transplanting to a soil-less substrate in a shaded greenhouse with more than a 98% survival rate. Three months after their establishment in the shaded greenhouse, 500 plants regenerated from stem explants were morphologically evaluated, from which five types of variants that had large, orbicular, elliptic, small, and lanceolate leaves were identified. Flow cytometry analysis of the variants along with the parent showed that they all had one identical peak, indicating that the variant lines, like the parent, were diploid. The mean nuclear DNA contents of the variant lines and their parent ranged from 4.90 to 4.99 pg 2C−1, which were not significantly different statistically. The results suggest that the regenerated plants have a stable ploidy level, and the regeneration method established in this study can be used for rapid propagation of ploidy-stable Aeschynanthus radicans.

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Acknowledgements

Authors thank Dr. Jaroslav Doležel at the Institute of Experimental Botany, Olomouc, Czech Republic and Dr. Thomas G. Ranney at North Carolina State University for providing Pisum sativum L. ‘Ctirad’ seeds, Wekiwa Gardens, Inc., Apopka, Florida for initially providing us with Aeschynanthus radicans ‘Mona Lisa’ plant materials, Terri Mellich for performing DNA flow cytometry analysis, and Russell Caldwell for critically reading this manuscript.

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  1. Department of Environmental Horticulture and Mid-Florida Research and Education Center, University of Florida, IFAS, 2725 Binion Road, Apopka, FL, 32703, USA

    Jin Cui, Jianjun Chen & Richard J. Henny

  2. College of Life Science, Nanjing Agricultural University, Nanjing, 210095, China

    Jin Cui

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  1. Jin Cui
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  2. Jianjun Chen
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Correspondence to Jianjun Chen.

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Editor: Praveen K. Saxena

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Cui, J., Chen, J. & Henny, R.J. Regeneration of Aeschynanthus radicans via direct somatic embryogenesis and analysis of regenerants with flow cytometry. In Vitro Cell.Dev.Biol.-Plant 45, 34–43 (2009). https://doi.org/10.1007/s11627-008-9147-9

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Keywords

  • Aeschynanthus
  • Flow cytometry
  • Medicinal plants
  • Ornamental foliage plants
  • Somatic embryogenesis