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

(1)从蕨类植物Lygodium japonicum孢子体幼叶中酶解分离原生质体，接种于含有NAA 2.7 um、BAP 2.2 um、0.6 M甘露醇和0.05 M蔗糖的MS培养基中。8 d内细胞发生分裂，观察到10-15个细胞。约2个月后，在无激素的培养基下，心形原体发育，同时具有卵母细胞和卵母细胞。(2)异孢子蕨类植物萨尔维尼亚的孢子体。由两种孢子（大孢子和小孢子）的雌配子体和雄配子体无菌共培养而得。从孢子植物幼叶中酶解分离原生质体。细胞在培养后6天内发生分裂。(3)采用组织培养法对异孢子植物马西莱（Marsilea quadrifolia L.）进行了离体增殖。(4)从百合小孢子细胞中分离纯化拓扑异构酶I，并对其进行了研究。

(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.