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In vitro culture may provide a suitable environment for selection of heavy-metal tolerant plantlets. Such clones of woody plants could be valuable material applicable to soil remediation. In in vitro culture conditions shoots of Daphne jasminea Sibth. & Sm. and Daphne tangutica Maxim. (Thymelaeaceae) were grown on media supplemented with 0.1, 0.5 and 1.0 mM lead nitrate. Level of lead bioaccumulation, growth parameters, content of photosynthetic pigments, and mineral status of cultured shoots were investigated. D. jasminea has grown vigorously on Pb2+-containing media, with growth tolerance index reaching 73–89%, depending on concentration applied, and the highest growth value was obtained in the presence of 1.0 mM lead nitrate. In vitro propagation of D. tangutica shoots was slightly inhibited by lead ions, however the growth tolerance index has increased up to 152% on medium with 1.0 mM Pb(NO3)2. In both studied species the highest content of accumulated lead, as well as the value of bioconcentration factor, were found in shoots grown on 0.1 mM lead nitrate. D. tangutica accumulated over two times as much lead in comparison with D. jasminea. Chlorophyll content in D. jasminea was not affected by applied lead nitrate doses, while in D. tangutica stimulation of chlorophyll, as well as carotenoid, synthesis occurred. In tested concentrations lead nitrate had no toxic effect on the level of shoot nutrition. Detected levels of essential and trace elements were still high enough to maintain undisturbed growth and development of cultured shoots. This is first report confirming the suitability of in vitro selection for obtaining of vigorous, proliferative, tolerant to elevated lead concentration shoots of ornamental Daphne species.
In this study, an attempt was made to investigate in vitro morphogenetic competence of three shrub species from the Thymelaeaceae family. The studied plant material originated from Russia, Greece and China, and the effectiveness of in vitro shoot formation and rhizogenesis of Daphne caucasica, D. jasminea, and D. tangutica was verified. The multiplication coefficient was compared for different propagation media. Medium composed of WPM mineral salts, MS microelements and a set of vitamins, supplemented with 1.0 mg dm-3 2iP, 0.1 mg dm-3 NAA, and 0.65 g dm-3 calcium gluconate, was appropriate for micropropagation of the tested genotypes. Shoot propagation in medium containing B5 vitamins and microelements was not as effective as on WPM/MS medium. The rooting phase, especially in D. tangutica, needs further optimization in order to reduce the costs associated with acclimatization of microplantlets obtained to in vivo conditions. After stabilization, the plants were successfully cultivated under greenhouse conditions.
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