The Role of Benzyladenine in the Differentiation of Tracheary Elements in Jerusalem Artichoke Tuber Explants Cultured in vitro

The role of benzyladenine (BA) in the differentiation of trachearyelements in Jerusalem artichoke (Helianthus tuberosus L.) tuberexplants was studied. For maximum differentiation of trachearyelements (25–30% of the cell population), treatment withoptimal concentrations of benzyladenine (5.0 mg dm^–3)in the presence of -naphthaleneacetic acid |NAA| (1.0 mg dm^–3)for the first 6 d was as effective as its continued presenceduring the entire 14 d period of study. A majority of the differentiatedtracheary element appeared between the 10th and 14th days ofculture. It was further observed that concentrations of activecytokinins in the tissue were considerably reduced within 2d after transfer from the BA-containing medium to a BA-freemedium. This was shown in three different ways: (1) monitoringthe amount of ethanol-soluble radioactivity at various timesafter transfer from |14C|-BA containing medium to BA-free medium;(2) bioassay of various cytokinin fractions from tissue extractseparated by thin layer chromatography; (3) indirect assay oftissue cytokinin activity through its interaction with abscisicacid for the promotion of auxin-induced cell division in thistissue. Both gibberellic acid (5.0 mg dm^–3) and abscisic acid(2–0 mg dm^–3) effectively inhibited the differentiationof tracheary elements even if provided after 6 d of pre-incubationin a high tracheid inducing medium. However, the appearanceof differentiated cells for the first 2 d after transfer wasnot significantly affected. A hypothetical scheme for the role of benzyladenine in the differentiationof tracheary elements in this tissue is discussed. It is suggestedthat during one or more critical cell divisions in the presenceof optimal levels of benzyladenine, a proportion of cells areinduced or committed for later differentiation into trachearyelements. The high concentrations of benzyladenine requiredduring induction are not needed during the intervening celldivisions, nor for the actual differentiation of the trachearyelements. Key words: Tracheary element differentiation, Jerusalem artichoke (Heliantlus tuberosus), Benzyladenine, Gibberellic acid, Abscisic acid