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

(1)从蕨类植物金鱼草孢子体幼叶中分离出原生质体，接种于含有NAA 2.7um、BAP 2.2um、甘露醇0.6M、蔗糖0.05M的MS培养基中。培养8d内细胞分裂，观察到10-15个细胞。约2个月后，在无激素培养条件下，心形原叶体发育成同时具心梗和颈卵器的组织。(2)异源蕨类植物丹参的孢子体。由两种类型的孢子(大孢子和小孢子)产生的雌、雄配子体共培养获得。用酶法从孢子体幼叶中分离出原生质体。(3)采用组织培养方法对异源四叶百合进行了体外增殖。(4)从百合小孢子细胞中分离、纯化并研究了拓扑异构酶I。

(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.