Plantlet regeneration from protoplast-derived embryogenic calli of Crocus cancellatus |
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Authors: | Karamian Roya Ebrahimzadeh Hassan |
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Institution: | (1) Department of Biology, Faculty of Science, Bu-Ali Sina University, Hamadan, Iran;(2) Department of Biology, Faculty of Sciences, University of Tehran, Iran |
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Abstract: | Plant regeneration from protoplast culture of Crocus cancellatus was investigated using regenerable embryogenic calli obtained from shoot meristem culture on LS (Linsmaier and Skoog, 1965)
medium containing 4 mg l−1 kinetin and 1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Protoplasts were isolated directly from embryogenic calli. The best protoplast growth
was found on those embedded in Ca-alginate beads and cultured with nurse cells in MS (Murashige and Skoog, 1962) medium supplemented
with 2 mg l−1 kinetin, 1 mg l−1 2,4-D and 100 mg l−1 ascorbic acid at 25 °C in darkness. After 4–5 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads,
but the protoplasts without immobilization in Ca-alginate beads showed very low cell division. Growth of the microcalli in
the medium with nurse cells was much better than in the medium without nurse cells. Transferring beads onto half strength
MS medium supplemented with 0.2 mg l−1 kinetin and 0.1 mg l−1 2,4-D, increased the growth of embryogenic calli. Somatic embryo development was observed either on half strength MS medium
growth regulator free or with 1 mg l−1 abscisic acid. Matured embryos germinated on half strength MS medium containing 25 mg l−1 of gibberelic acid. Plantlet formation was obtained on half strength MS medium containing 1 mg l−1 6-benzyladenine and 1 mg l−1 α-naphthaleneacetic acid at 20 °C in a 16/8 h light/dark cycle.
This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | Ca-alginate Crocus cancellatus nurse culture plantlet regeneration protoplast culture somatic embryogenesis |
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