Summary
Successful plant regeneration was achieved for the first time from hairy root-derived protoplasts of Hyoscyamus muticus. High yields (7 × 106 / g fresh weight) of protoplasts were isolated directly from the transformed roots of Hyoscyamus muticus using an enzyme mixture comprising 1 % macerozyme and 2 % cellulase in an osmoticum consisting of 0.2 M CaCl2 and 0.6 M mannitol. Protoplasts were first cultured in liquid NT/PRO I medium and further on semi-solid NT/PRO II agar medium. The procedure permits highly efficient formation of colonies. The plating efficiency varied from 1–9 %. The small individual colonies regenerated easily into shoots and roots at frequencies of 18 % and 70 %, respectively. The time required for the development of small plantlets from protoplasts was 8–11 weeks. The regenerated plants contained rolB from Ri-T-DNA and exhibited an altered phenotype compared to the control plants.
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Abbreviations
- BAP:
-
benzylaminopurine
- NAA:
-
naphthaleneacetic acid
- PCR:
-
Polymerase Chain Reaction
- fw:
-
fresh weight
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Communicated by N. Amrhein
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Sevón, N., Oksman-Caldentey, KM. & Hiltunen, R. Efficient plant regeneration from hairy root-derived protoplasts of Hyoscyamus muticus . Plant Cell Reports 14, 738–742 (1995). https://doi.org/10.1007/BF00232659
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DOI: https://doi.org/10.1007/BF00232659