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KMID : 0903619920330060425
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Journal of the Korean Society for Horticultural Science
1992 Volume.33 No. 6 p.425 ~ p.431
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Induction of Microspore - derived Embryos in Anther Culture of Raphanus sativus L.
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Abstract
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Microspore-derived embryos and plantlets were obtained successfully by anther culture of Raphanus sativus under the same condition as reported by Keller et al. Young floral buds in which the length of the anther is slightly longer than that of petal(petal : anther=3:4) were collected and rinsed with tap water and sterilized with 70% ethyl alcohol for 20 to 30 seconds. The bud were moved to 4.2% calcium hypochlorite emulsified with tween 20 for 10 minutes and washed with distilled water 4 times. The culture medium solid or liquid developed by Keller et al. for B. campestris was used together with that supplemented with 10% (w/v) coconut milk. The filaments of anthers were carefully removed under dissecting microscope and each 30 anthers were inoculated into petri-dish containing 25㎖ of medium. Petri-dish tightly sealed with paraffin film were incubated in 35℃ for 2 days in darkness and transferred to 25℃, where to be kept until regeneration. The emergence of embryoids per anther from the suture of the anther began to be seen on 15th day after inoculation. One to 10 embryos per anther were obtained in 30 to 35 days from culture. Proliferation of callus was not readily observed when filament tissue was completely removed and anther culture of Raphanus sativus was obtained the course of direct embryogenesis. Big difference in the anther culture response between varieties were recognized. It was also promotive in anther culture of radish to supplement the Keller´s medium with 2,4-D and NAA each 0.1㎎/ℓ.
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