The haploid embryoids development in anther culture of transformed tobacco (Nicotiana tabacum L.)

Induction of pollen embryogenesis was used for the test of cytokinin influence on the development of pollen grain. Gene for maize cytokinin-specific β-glucosidase were introduced into tobacco genome together with the methotrexate resistance marker (Brzobohatý et al. 1994). Anther culture of T1 generation of transformed Nicotiana tabacum L. plants revealed that high activity of β-glucosidase caused a reduced regeneration of pollen embryoids. The stages of pollen embryoid development were comparable with those of zygotic embryos. Optimal nutritive medium contained charcoal and chelated iron. The absence of iron in the initiation medium caused a distinct decrease of pollen embryoid regeneration