Abstract
Mesophyll protoplasts were isolated from leaf tissues of a diploid daylily (Hemerocallisx‘Red Magic’) by enzymatic digestion with a solution containing 0.5% Pectolyase Y-23, 0.1% Cellulase R-10, 0.1% Driselase, 0.6 M sorbitol and half-strength MS inorganic salts. When cultured on MS medium supplemented with 0.5 mg/l NAA and 0.5 mg/l BA, the protoplasts underwent sustained division to produce multicellular colonies. The optimal plating density for cell division was 0.5 × 105 protoplasts/ml. The highest plating efficiency was obtained in cultures grown in media solidified with 0.2% Gelrite. Under these conditions, formation of colonies occurred from 14% of cultured protoplasts. Calli were recovered from 9 colonies only after the cultures were treated with a conditioned medium. Intact plants were regenerated from protoplast-derived calli through organogenesis.
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Abbreviations
- BA:
-
6-benzylaminopurine
- FDA:
-
fluorescein diacetate
- GA3 :
-
gibberellic acid
- MS medium:
-
Murashige and Skoog (1962) medium
- NAA:
-
1-naphthaleneacetic acid
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Communicated by G. C. Phillips
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Ling, JT., Sauve, R.J. Isolation and culture of daylily mesophyll protoplasts. Plant Cell Reports 15, 293–296 (1995). https://doi.org/10.1007/BF00193739
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DOI: https://doi.org/10.1007/BF00193739