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Development of an in vitro protocol for a difficult-to-propagate endemic Australian dryland sedge species Mesomelaena pseudostygia (Cyperaceae)

  • Plant Tissue Culture
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Abstract

In vitro propagation for Mesomelaena pseudostygia a difficult-to-propagate dryland sedge species (Cyperaceae) endemic to Western Australia is described. Multiple avenues to in vitro propagation were investigated: shoot culture, organogenesis and somatic embryogenesis, with zygotic embryos as initiation material. The highest multiplication rate for shoots was 3.4 ± 1.0 after 6 wk on basal medium (1/2 strength Murashige and Skoog) with 2.5 μM kinetin and 0.5 μM 6-benzylaminopurine. Shoots achieved peak rooting (83%) following a pulse treatment on basal medium containing 10 μM indolebutyric acid and 2 μM α-naphthaleneacetic acid for 7 wk, followed by transfer to medium (without growth regulators) for a further 7 wk. Alternatively, in vitro grown shoots were pulse treated on basal medium with both 100 μM indolebutyric acid and 20 μM α-naphthaleneacetic acid for 1 wk then placed in Rockwool plugs (under propagation house conditions) for another 7 wk resulting in 63% root induction. Rooted plantlets were also successfully transferred to potting mixture either in Rockwool plugs or bare rooted and maintained in propagation house conditions with ≥95% survival after 7 wk. These results indicate that micropropagation of M. pseudostygia is feasible for small to medium scale restoration purposes. The highest frequency of callus induction was from cultured zygotic embryos on basal medium with 5 μM α-naphthaleneacetic acid, whereas 2,4-dichlorophenoxacetic acid (2 or 5 μM) produced the largest callus sizes. A low frequency of shoot regeneration occurred in zygotic callus tissues in basal medium treatments containing cytokinin (kinetin or thidiazuron at 1 μM). A small proportion (<20%) of zygotic embryo callus explants from 2,4-dichlorophenoxyacetic acid treatments were found to be embryogenic, firstly developing embryo-like structures after 2 wk on basal medium (minus plant growth hormones), that continued to develop with approximately one in twenty germinating after a further 4 wk on basal medium to form small plantlets. Further optimisation is needed to improve somatic embryogenesis efficiency for mass propagation.

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Acknowledgments

The authors acknowledge the assistance of Amanda Shade (Curator, Nursery) and other staff at the Kings Park and Botanic Gardens Nursery. SRT is presently supported by industry funding through Grange Resources and Cliffs Natural Resources.

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Authors and Affiliations

  1. Botanic Gardens and Parks Authority, Fraser Avenue, West Perth, WA, 6005, Australia

    Simin Lai, Akshay Menon, Shane Turner & Eric Bunn

  2. School of Plant Biology, Faculty of Natural and Agricultural Sciences, University of Western Australia, Crawley, WA, 6009, Australia

    Simin Lai, Shane Turner & Eric Bunn

  3. Western Australian Biomedical Research Institute, Curtin Health Innovation Research Institute, School of Biomedical Sciences, Curtin University, GPO Box U1987, Perth, WA, 6845, Australia

    Akshay Menon

  4. Melbourne School of Land and Environment, The University of Melbourne, Burnley Campus, Richmond, VIC, 3121, Australia

    Andrea Kodym

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  1. Simin Lai
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  2. Akshay Menon
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  3. Shane Turner
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  4. Andrea Kodym
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  5. Eric Bunn
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Correspondence to Eric Bunn.

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Editor: J. Forster

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Lai, S., Menon, A., Turner, S. et al. Development of an in vitro protocol for a difficult-to-propagate endemic Australian dryland sedge species Mesomelaena pseudostygia (Cyperaceae). In Vitro Cell.Dev.Biol.-Plant 50, 99–109 (2014). https://doi.org/10.1007/s11627-013-9542-8

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  • DOI: https://doi.org/10.1007/s11627-013-9542-8

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