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Authors: ALEXANDRU FIRA, DOINA CLAPA, ELENA RAKOSY-TICAN.
In vitro culture was successfully initiated by using seeds from cultivar Ningxia 1 as inocula cultured on Murashige and Skoog (1962) hormone-free medium, solidified with agar. The highest proliferation rates were obtained on the MS media with either 0.3 or 0.5 mg/l benzyl adenine, gelled with wheat starch as an agar alternative. The experimental treatments with 0.5 mg/l benzyl adenine ensured proliferation rates superior to the ones with 0.3 mg/l benzyl adenine, but the shoots obtained on MS + 0.3 mg/l benzyl adenine were longer and more robust. Also, the inoculation of large microcuttings onto the multiplication media ensured superior results regarding in vitro survival rates, the number of shoots regenerated / plantlet and the vigor of the plantlets. The microcuttings inserted vertically into the media yielded superior growth and multiplication as compared to the microcuttings placed horizontally on the surface of the media. The shoots regenerated in the multiplication stage could be used for cyclic in vitro multiplication. Explants either two centimetres or four centimetres in length proved to be effective. The non-rooted shoots resulting from the treatment with 0.3 mg/l benzyl adenine were either rooted in vitro on hormone-free MS medium gelled with starch or used for tests of ex vitro rooting and acclimatization. The optimal number of microcuttings in the in vitro rooting stage proved to be 40 explants/jar and the rooted plantlets were efficiently acclimatized ex vitro by three methods: float hydroculture in floating cell trays, floating perlite as well as Jiffy7 pellets. A successful alternative to in vitro rooting and subsequent ex vitro acclimatization was the direct ex vitro rooting and acclimatization in Jiffy7 pellets, in the same stage, using shoots excised from plantlets cultured in the multiplication stage. For direct ex vitro rooting and acclimatization, all other substrates we tested except for Jiffy7 pellets failed. Ex vitro rooting in floating perlite was stimulated by IBA but the technique was difficult and the results were not conclusive. After ex vitro acclimatization the resulting plants grew rapidly and vigorously. Acknowledgements. This work was possible with the financial support of the Sectoral Operational Programme for Human Resources Development 2007-2013, co-financed by the European Social Fund, under the project number POSDRU/107/1.5/S/76841 entitled “Modern Doctoral Studies: Internationalization and Interdisciplinarity”.
