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Plant Cell, Tissue and Organ Culture

, Volume 82, Issue 3, pp 281–288 | Cite as

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

  • Lin-Hui Li
  • Xian-Ping Wang
  • Wan-Ru Hou
  • Xiang-Lin Liu
  • Yi-Kun HeEmail author
Article

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.

Keywords

Atrichum undulatum (Hedw.) P. BeauvDNA uptake electroporation plant regeneration protoplast 

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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Copyright information

© Springer 2005

Authors and Affiliations

  • Lin-Hui Li
    • 1
    • 2
  • Xian-Ping Wang
    • 1
  • Wan-Ru Hou
    • 2
  • Xiang-Lin Liu
    • 1
  • Yi-Kun He
    • 1
    Email author
  1. 1.Department of BiologyCapital Normal UniversityBeijingChina
  2. 2.Department of BiologyChina West Normal UniversityNanChongChina

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