Maria Katarzyna Wojciechowicz, Małgorzata Anna Kikowska
Induction of multi-nucleate microspores in anther culture of Salix viminalis L.
Dendrobiology 2009, vol. 61: 55-64
Abstract: The aim of the work was to determine how the induction of androgenesis in selected genotypes of Salix viminalis is affected by thermal factors and medium composition. Anthers isolated from male clones of S. viminalis were pre-cultured at 4, 27 and 32°C for two to eight days in liquid Kyo medium with and without the addition of mannitol and on solid MS or WPM medium. The solid media were supplemented with different concentration of disaccharidesand various combinations of growth regulators including kinetin, 2iP, IAA and IBA. Multi-nucleate microspores indicative of androgenesis were observed in anthers pre-cultured for seven days at 4°C in liquid Kyo medium containing mannitol. The rate of androgenesis was higher when the anthers were transferred to solid modified MS medium containing high concentrations of sucrose and kinetin. In studied genotypes of basket willow early uninucleate microspores underwent sporophytic divisions. Ultrastructural observations showed differences in cellular arrangement of pre-stressed microspores. In Salix viminalis, microspores without starch grains and decreased number of lipid bodies were potentially androgenic cells.
Additional key words: mannitol, thermal shock, TEM, stress response