Abstract
The development of tissue-culture systems in duckweed has been limited to species of the genus Lemna. Here we report on a tissue-culture system (callus induction, callus growth and plant regeneration) for the rootless duckweed Wolffia arrhiza. We developed a two-step procedure for callus induction in Wolffia using Schenk & Hildebrandt (SH) medium containing glucose, mannitol and sorbitol. In the first stage, explants were precultivated in the presence of 5.0 mg l−1 2,4-dichlorophenoxyacetic acid and 0.5 mg l−1 N6-benzyladenine (BA) for 16 weeks. In the second stage, BA was replaced with 12.5 mg l−1 Picloram (PCL) for 4 weeks. The resulting callus could be maintained in vitro for a long time (about 1 year) with a relatively low concentration of PCL (4.0 mg l−1), or used to regenerate whole plants by transferring it to growth regulator-free SH medium. The protocols created for callus induction and regeneration were highly efficient at each stage (with the efficiency of 97 % for callus formation and 78 % for regeneration).
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