A fundamental research for genetic transformation via microprotoplast fusion in Liliaceous species

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Toward the application of microprotoplast fusion to genetic improvement of Liliaceous ornamental plants, 1)establishment of regenerable callus or cell suspension cultures, 2)development of protoplast culture systems, and 3)development of systems for isolating microprotoplasts, were examined.1)Highly regenerable callus cultures were established in Agapanthus praecox ssp. orientalis and Muscari armeniacum, and highly regenerable cell suspension cultures were established in Hosta sieboldiana.2)By utilizing callus cultures established, protoplast culture systems were developed in A. praecox ssp. orientalis and M. armeniacum. However, further studies are necessary to improve the efficiencies of callus formation from protoplasts and plant regeneration from protoplast-derived calluses.3)Two different systems for isolating microprotoplasts, one from partially synchronized cell suspension cultures of Hemerocallis hybrida(somatic microprotoplasts) and the other from developing microspores of Lilium longiflorum(gametic microprotoplasts) were developed. As compared with the somatic microprotoplast system, the gametic system seems to be more practical in Liliaceous ornamental plants.The systems developed in the present study may pave the way for the transfer of one or a few chromosomes via microprotoplast fusion in Liliaceous ornamental plants. Chromosome addition lines produced via microprotoplast fusion may contribute to chromosome studies as well as genetic improvement in these plants.