Abstract
A simple and efficient protocol for plant regeneration from mesophyll protoplasts ofNierembergia repens is described. The protoplasts divided in modified half-strength MS (/12 MS) medium containing benzylaminopurine (BA) and a-naphthaleneacetic acid (NAA) and formed visible colonies after 2 weeks which produced single adventitious shoots 4 weeks later. Plating efficiency (11.2%), percent colony formation (0.84%), and the number of shoot-forming colonies (368/dish) were highest in /12 MS containing 0.1 mg/l BA and 0.05 mg/l NAA. However, the percentage of colonies with shoot formation was highest (31.8%) in /12 containing 0.05 mg/l BA and 0.01 mg/l NAA. Almost all of the remaining colonies (97.5%) also regenerated shoots upon transfer onto MS medium containing 0.05 mg/l BA. The shoots with 2–3 leaves readily rooted 3–5 days after insertion in /12MS lacking plant growth regulators. Regenerated plantlets were easily established in soil 50 days after protoplast isolation. All the regenerants were normal and possessed diploid chromosome numbers.
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Abbreviations
- BA :
-
Benzylaminopurine
- FDA :
-
fluorescein diacetate
- MES :
-
2-(Morpholino)ethanesulfonic acid
- MS:
-
Murashige and Skoog (1962) medium
- /12MS :
-
half strength MS
- NAA:
-
α-naphthaleneacetic acid
- PGR :
-
plant growth regulator
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Communicated by J. Widholm
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Shizukawa, Y., Mii, M. A simple and efficient plant regeneration system for leaf protoplasts ofNierembergia repens by inducing single shoots on the microcolonies. Plant Cell Reports 16, 545–549 (1997). https://doi.org/10.1007/BF01142321
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DOI: https://doi.org/10.1007/BF01142321