Abstract
Leaf protoplasts of axenic shoot cultures of Lonicera nitida cv Maigrun underwent sustained division to give multicellular colonies (microcalli) on a modified, ammonium-free MS (Murashige & Skoog) medium containing 0.5 mg l-1 NAA (1-naphthaleneacetic acid), 1.0 mg l-1 BAP (6-benzylaminopurine) and 150 mg l-1 casein enzymatic hydrolysate. Callus was produced upon transfer of cell colonies to MS medium with 2.0 mg l-1 NAA and 0.2 mg l-1 BAP. About 110 days from isolation protoplast-derived shoots were regenerated on a half-strength MS medium with 0.01 mg l-1 NAA, 5.0 mg l-1 BAP, 0.5 mg l-1 zeatin and a complex mixture of group B vitamins. The replacement of such mixture by 250 mg l-1 casein enzymatic hydrolysate promoted rhizogenesis in calli, with shoot buds being subsequently regenerated from the protoplast-derived roots. Micropropagation of protoplast-derived shoots (of either origin) was difficult, due to a strong apical dominance, but could be accomplished by transferring single-node explants to half-strength MS medium with 1.5 mg l-1 BAP. Such shoots were, in turn, successfully rooted and transferred to the glasshouse where they completed acclimatization.
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Abbreviations
- BAP:
-
6-benzylaminopurine
- CPW:
-
Power et al. (1989) medium
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- FDA:
-
fluorescein diacetate
- F.P.E.:
-
final plating efficiency
- f.wt.:
-
fresh weight
- IAA:
-
4-indole-3yl-acetic acid
- IBA:
-
4-indole-3yl-butyric acid
- I.P.E.:
-
initial plating efficiency
- MES:
-
2-N-morpholinoethane sulfonic acid
- M.P.E.:
-
intermediate plating efficiency
- MS:
-
Murashige & Skoog (1962) medium
- NAA:
-
1-naphthaleneacetic acid
- PVP-10:
-
polyvinylpirrolidone
- Av MW 10,000, TIBA:
-
2,3,5-tri-iodobenzoic acid
- Z:
-
zeatin
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Ochatt, S.J. Requirements for plant regeneration from protoplasts of the shrubby ornamental honeysuckle, Lonicera nitida cv. Maigrun. Plant Cell Tiss Organ Cult 25, 161–167 (1991). https://doi.org/10.1007/BF00042188
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DOI: https://doi.org/10.1007/BF00042188