摘要
以杉木（Ｃｕｎｎｉｎｇｈａｍｉａｌａｎｃｅｏｌａｔａ（Ｌａｍｂ．）Ｈｏｏｋ．）传粉后的胚珠为材料，分离获得了生活的雌配子和雄配子。当花粉管进入雌配子体组织后，在颈卵器上方形成膨大的精原细胞或成对的精细胞。解剖出的精细胞呈ＦＤＡ正反应，荧光增白剂染色表明精细胞具有细胞壁。雌配子体在酶液作用下可同时释放出多个卵细胞和精细胞。游离出的卵细胞呈ＦＤＡ正反应，近圆形，内含１～２个大液泡。用ＰＥＧ方法诱导卵细胞与卵细胞，卵细胞与精细胞的融合，只获得了相互粘连的两个原生质体。
Viable male and female gametes were isolated from pollinated ovules of Cunninghamia lanceolata (Lamb.) Hook. Prior to their penetration into the female gametophyte, the pollen tubes were drawn out from the nucellus. The isolated pollen tubes were branched and one of them became swollen. An enlarged spermatogenous cell and subsquently a pair of sperm cells were formed as the pollen tube reached the regions over and against the archegonia. The sperm cells were released from the pollen tubes manually with the use of a stereomicroscope. The positive FDA reaction gave evidence of the sperm cells viability, and the Fluorescent Brightener 28 positive demonstrated the presence of cell wall. The egg cells were enzymatically isolated from the female gametophytes.The isolated egg cells were spherical, contained 1 to 2 large and many small vacuoles. FDA test showed the egg cells were viable, and the viability sustained for 8 days in 2 to 4 ℃ without any protectants. Fusion between single pair of male and female gametic protoplasts was attempted with PEG method, but only adhesion of the two was obtained.
基金
国家自然科学基金
关键词
分离
卵细胞
精细胞
融合
杉木
Isolation, Male gamete, Female gamete, Cunninghamia lanceolata
