Somatic embryogenesis in Cryptomeria japonica was initiated at a relatively high frequency from immature seeds collected from the end of June to mid -July. Induction of embryogenic cultures was possible on media with or without plant growth regulators, and the initiation frequency varied from 5 to 16%. Embryogenic cell lines have been maintained and proliferated for more than 2 years in solid and liquid media. For long-term storage embryogenic cells were cryopreserved using a simple freezing method. Cotyledonary embryos were obtained mostly on maturation media containing abscisic acid (ABA) and polyethylene glycol (PEG) as osmotic agent, however, the plant conversion rate was still low. Plants regenerated from somatic embryos continued growing in a greenhouse. Furthermore, a procedure for the individual culture of protoplasts isolates from embryonal masses, and an approach for microprojectile bombardment-mediated transformation using pIPT and pMAT vectors was also described.
This report describes the successful plant regeneration via somatic embryogenesis from immature zygotic embryos of Cryptomeria japonica D. Don. For the induction of embryogenic tissue, we determined that the optimal medium contained N6-benzyladenine and 2,4-dichlorophenoxyacetic acid. Immature zygotic embryos that were collected at the end of June yielded embryogenic tissue at the highest frequency. Embryogenic tissues that had proliferated in liquid medium included small and loosely packed cells and elongating or elongated cells. We used ten cell lines to determine the optimal medium for the development of somatic embryos. Induced somatic embryos germinated with synchronous sprouting of cotyledons, hypocotyls and roots. Gibberellin A3 in the germination medium had a positive effect on both the elongation of hypocotyls and the survival of seedlings. The frequencies of induction and germination of somatic embryos differed among the cell lines examined. Most of the seedlings grew normally. This system of somatic embryogenesis required 4-5 months for the regeneration of C. japonica plantlets from immature zygotic embryos.
This report describes the efficient plant regeneration of Chamaecyparis obtusa Sieb et Zucc. via somatic embryogenesis. Embryogenic cultures were initiated from megagametophytes containing immature zygotic embryos. Embryogenic cultures were maintained and proliferated by 2-3-week interval subcultures in medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. High maturation frequencies of cotyledonary embryos were obtained on maturation medium containing maltose, polyethylene glycol, activated charcoal, and abscisic acid. Somatic embryos germinated readily after transfer to plant growth regulator-free medium. Growth of regenerated emblings has been monitored in a greenhouse.
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