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An efficient protocol for plant regeneration from protoplasts of the moss Atrichum undulatum P. Beauv in vitro

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Abstract

A system for plant regeneration from protoplasts of the moss, Atrichum undulatum (Hedw.) P. Beauv. in vitro, is first reported. Viable protoplasts were isolated at about 9 × 105 protoplasts g−1 fresh weight from 10 to 18 days protonemata. For regeneration of protoplasts, viable protoplasts were cultured in liquid–solid medium containing surface liquid medium MS (0.4 M mannitol) and subnatant solid medium Benecke (0.3 M mannitol) at 20 °C under a 16-h photoperiod white light after 12 h preculture in darkness at 20 °C. The great majority of protoplasts follow a regenerative sequence: formation of asymmetric cells in 2–3 days; division of the asymmetric cells to 2–3 cells in 4–5 days, and further develop to produce a new chloronemal filament in 15 days. Juvenile gametophyte can be visible in 20 days. The plating ratio of cell cluster regenerated from protoplasts reaches up to 45%. Transient expression experiments indicate the electroporation uptake of DNA is possible.

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Abbreviations

FDA:

fluorescein diacetate

FW:

fresh weight

Gus:

β-glucuronidase

MES:

2-(N-morpholino) ethanesulphonic acid

MS:

Murashinge and Skoog

PEG:

polyethlence glycol

PE:

plating efficiency

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Authors and Affiliations

  1. Department of Biology, Capital Normal University, Beijing, 10037, China

    Lin-Hui Li, Xian-Ping Wang, Xiang-Lin Liu & Yi-Kun He

  2. Department of Biology, China West Normal University, NanChong, 637002, Sichuan, China

    Lin-Hui Li & Wan-Ru Hou

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  1. Lin-Hui Li
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  2. Xian-Ping Wang
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  3. Wan-Ru Hou
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  4. Xiang-Lin Liu
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  5. Yi-Kun He
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Correspondence to Yi-Kun He.

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Li, LH., Wang, XP., Hou, WR. et al. An efficient protocol for plant regeneration from protoplasts of the moss Atrichum undulatum P. Beauv in vitro. Plant Cell Tiss Organ Cult 82, 281–288 (2005). https://doi.org/10.1007/s11240-005-1370-z

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  • DOI: https://doi.org/10.1007/s11240-005-1370-z

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