Successive Culture and Organogensis of haploid plants in Equisetum arvense L.

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1. We have established cultured gametopluyte tissue originated from a spore of Equisetum arvense L. on the Murashige & Skoog's medium added 3% sucrose and 0.8% agar. It is very rare case to establish and subculture of haploid gametophyte in aseptic condition for a long time.2. Shoot formation occured by the effects of a low concentration of 6-benzil adenine (6-BA) on th gametophyte, however Kinetin, Zeatin and 6-PU have a little effects as compared with 6-BA.3. All tested phytohormones, IAA, NAA, ABA, 2,4-D and GA3, showed some inhibiting effects to shoot fromation.4. Morphological observation of the shoot formation was observed by routine method of light microscope, and transmit- and scanning electron microscope. Apogamy from a epidermal cell of cultured gametophyte was confirmed by these observation.5. In the light, differentiation of antheridia occured in the subcultured gametophyte on MS-medium added 3% sucrose. Many sperm cells were observed in the antheridia during subculture for a long period.6. Modification of sucrose concentration in the medium was required for archegonium formation on the cultured gametophyte. The arechegonia were observed after transplantation from 3% sucrose medium with 6-BA to 0.5%. It seems that the effect is specific in sucrose, because the effect was not appeared with many sugars other than sucrose.7. Protoplasts were isolated from cultured gametophyte tissue by digestive enzymes, and they were cultivated for long term. Regeneration of cell wall, cell division of the protoplasts and gametophyte regeneration were observed.