DEVELOPMENT OF PRODUCTION PROCESS OF ARTIFICIAL SEEDS USING CELL AGGREGATES DERIVED FROM PLANT HAIRY ROOTS

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The aim of the present research is to develop artificial seeds for the mass propagation of useful plant variants. This research is comprised of the following parts : 1) process of cell aggregate (referred to as CA) production from hairy roots, 2) preparation of artificial seeds by CA encapsulation, and 3) process of root emergence and plant development from artificial seeds. In each process, the hairy root cells were used because of their genetic stability and fast growth rate, and the following results were obtained :1. The medium with phytohormones, namely, BAP (6-benzylaminopurine) and NAA (1-naphthaleneacetic acid) was effective for the formation of CA from horseradish hairy roots. It was demonstrated that these CA were useful as artificial embryos. The synergistic effects of BAP and NAA on the CA formation were also clarified with a kinetic model.2. During encapsulation of CA in alginate gel beads, it was shown that the regenetation potential of CA could be enhanced by NAA treatment. To improve the storage potential of the artificial seeds, the application of coats on the alginate gel surface was considered, and paraffin was found to be the most suitable among the various materials tested.3. It was observed that the root emergence from CA could be expressed as a normal distribution function of time, and the kinetics of CA regeneration was analyzed. Moreover, it was shown that the regenerating CA subsequently developed into whole plants upon transfer under light condition. On the other hand, the relation between regeneration effeciency and permeability of paraffin coat membrane to oxygen was elucidated, and the regeneration potential of CAA after 60 days of storage was verified.It is considered that the above findings could contribute to the advancement of artificial seed technology for the preservation and mass propagation of useful plants.