Maciej Zenkteler

Cytological analysis of hybrid embryos of intergeneric crosses between Salix viminalis and Populus species.

Intergeneric hybridisation between Salix viminalis L. as the female and four Populus species (Populus trichocarpa, P. tremula, P. × canadensis and P. simonii) as male pollen donors was performed by in vitro stigma pollination. To overcome postzygotic barriers, transfer of hybrid embryos to new medium is necessary. We carried out detailed ultrastructural analyses to establish: (i) at which stage of embryo development the first signs of programmed cell death (PCD) could be detected; and (ii) at which stage the lack of serious or irreversible changes guaranteed that advanced development of hybrid plants could occur after embryo rescue. Transmission electron microscopy and confocal laser scanning microscopy revealed the presence of both developing and degenerating embryos. Developing globular, heart-shaped, and early cotyledonary embryos contained cells of correct ultrastructure. The only sign of intergeneric hybridisation was a delay in development for a few days, in comparison with control embryos. The earliest indicators of embryo degeneration were noted at 9 days after pollination (DAP). The most common indicators were excessive embryo vacuolisation, which was characterised by a large number of vesicles and formation of small vacuoles, as well as enlarged central vacuoles. Extended plastid thylakoids, folding of the cell wall, and autophagosomes were observed. Our detailed investigation of PCD in hybrid embryos enabled us to conclude that the embryo rescue technique was most effective in intergeneric willow × poplar crosses if applied between 9 and 16 DAP.

The de novo Formation of Buds and Plantlets from Various Explants of Ailanthus altissima Mill. Cultured in vitro

The hypocotyls, cotyledons, leaf blades, whole leaves and petioles of seedlings ofAilanthus altissima are capable of producing callus and budsin vitro. Buds and callus were also obtained from whole leaves and internodes of 2-years old plantlets grownin vitro. From the calli buds differentiated and later, both from buds developing directly without a callus phase and alsovia a callus phase, well developed shoots were formed. The cultures were mainained on MS medium in 2 combinations: A) IAA - 0.2 mg 1−1, BAP - 1 mg 1−1, GA3 - 0.5 mg 1−1, thiamine - 4 mg 1−1 and sucrose 3 %; B) BAP - 0.5 mg 1−1, IAA - 1 mg 1−1, casein hydrolysate 400 mg 1−1, thiamine 4 mg 1−1 and sucrose 3 %. Excised shoots, which had developedde novo in culture, produced roots when incubated on the basic mineral medium of MS with the addition of IAA. The regenerative potential ofA. altissima is very high and this woody species seems to be an ideal object for various morphogenetic studies.