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A high-frequency cyclic secondary somatic embryogenesis system for Cyclamen persicum Mill

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Abstract

In this study, a high frequency cyclic secondary somatic embryogenesis system was established for Cyclamen persicum. Moreover, the influence of primary somatic embryos (PSEs) at different stages of development, number of passages, abscisic acid (ABA), and sucrose concentrations on secondary somatic embryogenesis were investigated. Primary somatic embryos, at different stages of development, were incubated on an induction medium consisting of half-strength Murashige and Skoog (MS) (Physiol Plant 15:473–497, 1962), 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.2 mg/l 6-benzyladenine (BA). These initial PSE cultures along with calli were transferred to a plant growth regulator (PGR)-free medium to promote secondary somatic embryo (SSE) development. Embryogenic calli were induced at a frequency >90% from all PSE cultures. Both morphology and size of PSEs influenced embryogenic competence. Large-sized globular embryos (GEs) yielded the highest number of SSEs followed by small- and large-sized torpedo-stage embryos (TEs). Passages of induced calli obtained from large-sized GEs had no effects on frequency of secondary embryogenesis; however, they significantly influenced SSE production. Calli from the first passage exhibited the highest competence for secondary embryogenesis, 100% frequency of SSEs, producing 3,306 and 1,296 SSEs per gram fresh weight callus and embryo, respectively. Incubating embryogenic calli derived from large-sized primary GEs in the first passage on a medium containing 30 g/l sucrose, and supplemented with either 0.5 or 1.0 mg/l abscisic acid (ABA) increased the frequency of TEs, consequently enhancing normal embryo development of SSEs. Transfer of such embryogenic calli to a PGR-free medium containing 60 g/l sucrose was optimal for SSE induction, producing 3,204 SSEs per gram callus and 8% TEs.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

ABA:

Abscisic acid

BA:

6-Benzyladenine

GEs:

Globular embryos

MS:

Murashige and Skoog’s medium

NTEs:

Non-polar tuber embryos

PGRs:

Plant growth regulators

PSEs:

Primary somatic embryos

SEs:

Somatic embryos

SSEs:

Secondary somatic embryos

TEs:

Torpedo-stage embryos

TWEs:

Twin embryos

UBEs:

Unipolar tuber-bud embryos

UTEs:

Unipolar tuber-root embryos

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Acknowledgments

The authors thank Dr. Jian-cheng Song from School of Biological Sciences, University of Canterbury, New Zealand and Dr. Rong-shuang Lin from University of Maryland, College Park, USA for their assistance in editing and proofreading the manuscripts. This project was supported by Key Scientific and Technological Projects of Shandong province (Contract No. 2004GG4202010 and 2007GG20002019) and Natural Science Foundation of Shandong province (Grant No. Y2007D07).

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Authors and Affiliations

  1. College of Marine Life Sciences, Division of Life Science and Technology, Ocean University of China, Qingdao, 266003, Shandong, China

    Cui Rong You & Ting Jun Fan

  2. College of Life Sciences, Yantai University, Yantai, 264005, Shandong, China

    Cui Rong You, Xue Qin Gong, Fu Hua Bian, Li Kun Liang & Fu Ning Qu

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  1. Cui Rong You
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  2. Ting Jun Fan
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Correspondence to Fu Ning Qu.

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You, C.R., Fan, T.J., Gong, X.Q. et al. A high-frequency cyclic secondary somatic embryogenesis system for Cyclamen persicum Mill. Plant Cell Tiss Organ Cult 107, 233–242 (2011). https://doi.org/10.1007/s11240-011-9974-y

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