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

异孢子蕨类植物雌雄细胞分化的研究

• 批准号: 02640538
• 负责人: MAEDA Masuo
• 金额: $ 1.15万
• 依托单位: Fukui University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02640538/
• 关键词: ferns; heterospory; sex organ differentiation; cell culture; protoplast; female and male gametophytes; meiosis; microsporocytes; 異型胞子; 顕微螢光測光

(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.

（1）从海金沙孢子体幼叶中酶解分离的原生质体接种于MS + NAA2.7 μ m，BAP2.2 μ m，0.6M甘露醇，0.05M蔗糖的培养基中。细胞分裂在培养8天内发生，观察到10-15个细胞。约2个月后，在无菌培养基中，心形原叶体发育，同时具有精子器和颈卵器。（2）异孢蕨Salvinia natans All.通过共培养来自两种类型孢子（大孢子和小孢子）的雌配子体和雄配子体而无菌获得。从孢子体幼叶中酶解分离原生质体。细胞分裂在培养的6天内发生。(3)In对四叶散菜进行离体增殖，异孢子生殖。（4）从百合小孢子母细胞中分离纯化了拓扑异构酶I。

项目成果

M.Maeda,Y.Sugimoto,M.Nakamura K.Masuda,H.Kaneko,and M.Sugai: "Division and gametophytic tissue formation from protoplasts of young sporophytes in fern Lygodium japonicum" Plant Cell Reports. 9. 113-116 (1990)

M.Maeda、Y.Sugimoto、M.Nakamura K.Masuda、H.Kaneko 和 M.Sugai：“蕨类海金沙幼孢子体原生质体的分裂和配子体组织形成”植物细胞报告。

前田 桝夫・中村 三千代: "シダ類の組織培養 ー 形態形成実験系としての利用 ー (特集・植物培養細胞の利用:菌類からシダ植物まで)" 遺伝 (裳華房), 43-47 (1990)

Masuo Maeda 和 Michiyo Nakamura：“蕨类植物的组织培养 - 用作形态发生实验系统 -（特色：培养植物细胞的使用：从真菌到蕨类植物）” 遗传学 (Shokabo)，43-47 (1990)

Michiyo Nakamura: "Induction and establishment of haploidic and diploidic cell lines from a fern,Anemiya phyllitidis" Plant Cell Reports. (1992)

Michiyo Nakamura：“从蕨类植物 Anemiya phyllitidis 中诱导和建立单倍体和二倍体细胞系”《植物细胞报告》。

Ohyama,T.,S.Tabata,T.Kobayashi,S.Sato,M.Maeda,and Y.Hotta: "Purification and preliminary characterization of DNA topoisomerase I from lily microsporocytes" Plant Science.

Ohyama,T.,S.Tabata,T.Kobayashi,S.Sato,M.Maeda,and Y.Hotta：“百合小孢子母细胞 DNA 拓扑异构酶 I 的纯化和初步表征”植物科学。

T. Ohyama, S. Tabata, To Kobayashi, S. Sato, M. Maeda, and Y. Hotta: "Purification and preliminary characterization of DNA topoisomerase I from lily microsporocytes" Plant Science.

T. Ohyama、S. Tabata、To Kobayashi、S. Sato、M. Maeda 和 Y. Hotta：“百合小孢子母细胞 DNA 拓扑异构酶 I 的纯化和初步表征”《植物科学》。