Abstract
Cibotium barometz is an endangered tree fern, used both as ornamental plant and traditional Chinese medicinal plant. In this study, an effective in vitro propagation protocol was obtained through formation of green globular bodies (GGBs) from in vitro juvenile sporophytes. The effect of plant growth regulators (PGRs) on GGB induction and multiplication, as well as mineral salt concentration and active charcoal (AC) on plantlet regeneration from GGBs was evaluated. Thidiazuron (TDZ; 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea) had a significant effect on GGB induction and multiplication (P < 0.001), while a-naphthaleneacetic acid (NAA) did not (P > 0.05). GGB induction rate was above 80 % on 1/2 Murashige and Skoog (MS) media supplemented with TDZ (1.0 mg L− 1) and NAA (0.1, 0.3 or 0.5 mg L− 1). The same media were also optimal for GGB multiplication. GGBs cultured on 1/4 MS media supplemented with 0.1 or 0.2 % (w/v) AC showed a high rate of GGB development into plantlets above 90 %. 1/2 MS media supplemented with 0.1 or 0.2 % AC were the most effective for plantlet growth. Regenerated plantlets were successfully acclimatized (80 %) in greenhouse conditions. Morphological and histological analysis revealed that C. barometz GGBs was a yellow-green globular structure composed of the single GGB with meristems and hair-like structures, and new single GGBs were initiated from the epidermal cells of meristem zone.
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Abbreviations
- 2iP:
-
N6-(2-Isopentenyl) adenine
- AC:
-
Active charcoal
- BA:
-
N6-benzyladenine
- CITES:
-
Convention for international trade on endangered species
- GGBs:
-
Green globular bodies
- LSD:
-
Least significant difference
- MS:
-
Murashige and Skoog (1962)
- NAA:
-
a-Naphthaleneacetic acid
- PGR:
-
Plant growth regulator
- PLBs:
-
Protocorm-like bodies
- TDZ:
-
Thidiazuron (1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea)
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Acknowledgments
We thank Anthony E. Baniaga (Department of Ecology & Evolutionary Biology, University of Arizona) and Haifei Yan (South China Botanical Garden, Chinese Academy of Sciences) for their help in revising the paper. This work was financially supported by the National Engineering Research Center for Ornamental Horticulture (2012FU125X10), Yunnan Foundation Research Projects for Application (2014FD066), Yunnan Science and Technology Projects (2014RA053), Yunnan Biodiversity Conservation Projects, Innovation Team of Yunnan Academy of Agricultural Sciences (2014CZYY019), PR China.
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Yu, R., Zhang, G., Li, H. et al. In vitro propagation of the endangered tree fern Cibotium barometz through formation of green globular bodies. Plant Cell Tiss Organ Cult 128, 369–379 (2017). https://doi.org/10.1007/s11240-016-1116-0
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DOI: https://doi.org/10.1007/s11240-016-1116-0