Direct organogenesis provides an efficient means of propagating plants in vitro without the necessity of growing callus cultures. Shoot regeneration was achieved from bud explants of Bienertia sinuspersici on Murashige and Skoog's media supplemented with 6-benzyladenine (BA). The frequency of regeneration varied based on the concentration of BA used. The highest rate of new shoot induction was observed when explants were cultured on media containing 4.4 lM BA after three weeks. Shoots were successfully elongated on media containing gibberellic acid with the highest shoot elongation observed at 2.9 lM. Elongated shoots developed adventitious roots in media without plant growth regulator and were subsequently transplanted to soil. Light microscopic analysis of cross sections of leaves from regenerated shoots showed similar intracellular cytoplasmic compartmentalization of organelles in chlorenchyma cells compared to those observed in leaves grown under greenhouse conditions. Moreover, Western blot analysis detected high expression of key C 4 photosynthetic enzymes, suggesting the C 4 cycle is maintained in the single cell system. We have developed a procedure for whole plant regeneration via direct organogenesis in B. sinuspersici.
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