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Direct and indirect shoot and bulblet regeneration from cultured leaf explants of Lilium pumilum, an endangered species

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Abstract

Alternative methods of in vitro cloning that involve both adventitious (direct) and callus intermediate (indirect) pathways were investigated for the endangered species Lilium pumilum. Plantlet regeneration was obtained from leaf explants, cultured on Murashige and Skoog (MS) basal medium supplemented with various combinations of auxins and cytokinins at different concentrations. About 30% of the explants directly formed adventitious shoots on MS medium containing 8.88 μM 6-benzyladenine (BA) and 2.69 μM α-naphthaleneacetic acid (NAA). For production of regenerable callus, callus formation followed by shoot induction was best when explants were initially cultured on MS medium supplemented with 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Regenerable calli were yellow or purple and readily regenerated shoots when subcultured onto MS medium containing 2.22 μM BA and 1.61 μM NAA. About 78% of the calli were able to produce adventitious shoots. Shoots were rooted on half-strength MS medium supplemented with 1.34 μM NAA and were successfully acclimatized to greenhouse conditions. This report describes an efficient method for the in vitro multiplication of whole plants from leaf explants of the endangered species L. pumilum.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (NSFC) (31070616), the Natural Science Foundation of Heilong Jiang Province (No. C200714), and the Youth Science Foundation of Harbin City (No. 2008RFQXN007).

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Authors and Affiliations

  1. Key Laboratory of Saline-Alkali Vegetation Ecology Restoration in Oil Field (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin, 150040, China

    Shumei Jin, Ji Wang, Dan Sun & Shengkui Liu

  2. Texas A&M AgriLife Research Center, Texas A&M University System, 17360 Coit Road, Dallas, TX, 75252, USA

    Xinwang Wang & A. D. Genovesi

  3. Maize Research Institute of Heilongjiang Academy of Agricultural Sciences, Harbin, 150086, China

    Guoliang Li

Authors
  1. Shumei Jin
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  2. Ji Wang
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  3. Xinwang Wang
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  4. Dan Sun
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  5. Guoliang Li
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  6. A. D. Genovesi
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  7. Shengkui Liu
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Corresponding authors

Correspondence to Xinwang Wang or Shengkui Liu.

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Editor: Wagner Campos Otoni

S. Jin, J. Wang, and X. Wang contributed equally to this work.

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Jin, S., Wang, J., Wang, X. et al. Direct and indirect shoot and bulblet regeneration from cultured leaf explants of Lilium pumilum, an endangered species. In Vitro Cell.Dev.Biol.-Plant 50, 69–75 (2014). https://doi.org/10.1007/s11627-013-9573-1

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  • DOI: https://doi.org/10.1007/s11627-013-9573-1

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