Abstract
Protoplasts ofVicia narbonensis isolated from epicotyls and shoot tips of etiolated seedlings were embedded in 1.4% sodium-alginate at a final density of 2.5×105 protoplasts/ml and cultivated in Kao and Michayluk-medium containing 0.5 mg/I of each of 2,4- dichlorophenoxyacetic acid, naphthylacetic acid and 6 -benzylaminopurine. A division frequency of 36% and a plating efficiency of 0.40–0.5% were obtained. Six weeks after embedding, protoplast-derived calluses were transferred onto gelrite-solidified Murashige and Skoog-media containing various growth regulators. Regeneration of plants was achieved via two morphologically distinguishable pathways. A two step protocol (initially on medium with a high auxin concentration followed by a culture phase with lowered auxin amount) was used to regenerate somatic embryos, whereas cultivation on medium containing thidiazuron and naphthylacetic acid resulted in shoot morphogenesis. Mature plants were recovered from both somatic embryos as well as from thidiazuron-induced shoots.
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Abbreviations
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- BA:
-
6 -benzylaminopurine
- CH:
-
casein hydrolysate
- FDA:
-
fluorescein diacetate
- IBA:
-
indole-3-butyric acid
- KM:
-
Kao and Michayluk's medium (1975)
- MS:
-
Murashige and Skoog's medium (1962)
- NAA:
-
naphthylacetic acid
- TDZ:
-
thidiazuron
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Communicated by H. Lörz
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Tegeder, M., Kohn, H., Nibbe, M. et al. Plant regeneration from protoplasts ofVicia narbonensis via somatic embryogenesis and shoot organogenesis. Plant Cell Reports 16, 22–25 (1996). https://doi.org/10.1007/BF01275442
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DOI: https://doi.org/10.1007/BF01275442