A simple and efficient plant regeneration system for leaf protoplasts and Nierembergia repens by inducing single shoots on the microcolonies
1997
Shizukawa, Y. | Mii, M.
A simple and efficient protocol for plant regeneration from mesophyll protoplasts of Nierembergia repens is described. The protoplasts divided in modified half- strength MS (1/2 MS) medium containing benzylaminopurine (BA) and alpha-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 1/2 MS containing 0.1 mg/l NAA and 0.05 mg/l NAA. However, the percentage of colonies with shoot formation was highest (31.8%) in 1/2 MS 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 1/2 MS 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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