Effects of Chilling, Anther Preculture and Growth Regulators on Embryogenesis in Isolated Pollen Culture of Nicotiana rustica L.

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Pollen culture of Nicotiana rustica was performed by using 3 tages of pollen as the experimeutal materials. Freshly isolated pollen failed to form embryoid regardless of the presence of NAA and BA. Embryoids were produced from pollen isolated from anthers precultured on agar medium for 4 days or liberated. from anthers floated on liquid medium for 6 days. Prolonged preculture, however, reduced embryoid formation in both methods. Presence of NAA and BA in the media inhibited the induction of embryogenesis even at extremely low concentrations. Chilling pretreatment of flower buds at 5°C for 4 days remarkably increased embryoid formation. The inhibitory effect of prolonged preculture on liquid medium medium was cancelled by the chilling pretreatment, but not the case in the preculture on agar medium. By using the method with preculture on liquid medium, embryoid formation occurred in both uninucleate and mitotic stages but not in the early binucleate stage where high androgenic response, although, was observed in anther culture.
日本育種学会の論文