Study on the differentiation of sex organs using female and male cells in heterosporous ferns

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(1)Protoplasts isolated enzymatically from juvenile leaflets of sporophytes in fern Lygodium japonicum were inoculated in a MS medium containing NAA 2.7 um, BAP 2.2 um, 0.6 M mannitol, and 0.05 M sucrose. Cell division took place within 8 days of culture with 10-15 cells were observed. About 2 months later, cordate prothallia developed bearing both antheridia and archegonia under hormone-free medium.(2)Sporophytes in heterosporous fern Salvinia natans All. were aseptically obtained by co-culture of female and male gametophytes derived from two types of spores (megaspores and microspores). Protoplasts were isolated enzymatically from juvenile leaflets of sporophytes. Cell division took place within 6 days of culture.(3)In vitro multiplication of Marsilea quadrifolia L., a heterospory, was carried out by tissue culture method.(4)Topoisomerase I was isolated, purified, and studied from microsporocytes of Lilium.