Abstract
Leaf mesophyll protoplasts ofDianthus superbus were cultured at a density of 5 × 104 protoplasts/ml and divided at about 18% plating efficiency in MS liquid medium supplemented with 0.5 mg/L BAP, 2.0 mg/L NAA and 9% mannitol after 2 weeks. Protocolonies formed after 3 to 4 weeks of culture in the dark at 27°C. These colonies were transferred to continuous illumination (21.5 μE m−2 sec−1) for 2 weeks where most of the colonies divided to form microcalli, about 2 mm in diameter. Subsequently, green microcalli were transferred to MS solidified medium with 2.0 mg/L 2,4-D that induced shoot-forming calli after 4 weeks. These calli were transferred onto N6-2 medium containing 0.1 mg/L 2,4-D, 0.1 mg/L NAA, 2.0 mg/L kinetin and 2.0 g/L casein hydrolysate and were cultured under light. After 5 weeks the calli gave rise to multiple shoots (10 to 15 per callus). Upon transfer to MS medium containing 2.0 mg/L NAA, individual shoots were rooted in 4 weeks. The regenerants were successfully transplanted into potting soil.
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Abbreviations
- MS:
-
Murashige and Skoog
- BAP:
-
6-benzylaminopurine
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- NAA:
-
1-naphthaleneacetic acid
- N6 :
-
Chu basal salt mixture
- MES:
-
2-N-morpholinoethanesulfonic acid
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Communicated by I. K. Vasil
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Kim, J.C., Lee, E.A. Plant regeneration from mesophyll protoplasts ofDianthus superbus . Plant Cell Reports 16, 18–21 (1996). https://doi.org/10.1007/BF01275441
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DOI: https://doi.org/10.1007/BF01275441