摘要
用PEG法诱导了玉帘(Zephyranthes candida)、石蒜(Lycoris radiata)、黄花菜(Hemerocallisminor)、朱顶红(Hippeastrum vittata)和鸢尾(Iris tectorum)五种植物的同种生殖细胞之间、玉帘与石蒜异种生殖细胞之间、石蒜生殖细胞与其花瓣原生质体之间、石蒜生殖细胞与油菜下胚轴原生质体之间的融合,获得了含二至多个细胞核的同核体和异核体。其中石蒜生殖细胞的同核体还进一步发生了细胞核的融合。用与DNA特异结合的荧光染料DAPI预先标记的方法,可以在融合体中显示生殖细胞核。FDA荧光鉴定表明融合体是生活的。还研究了若干因素对玉帘生殖细胞之间融合的影响。
The generative cells used for fusion experiments were isolated from pollen grains of Zephyranthes Candida and Lycoris radiata by '2-step osmotic shock' and from those of Hippeastrum vittata,Hemerocallis minor and Iris tectorum by 'weak enzyme treatment' as reported previously.Using PEG method,fusions have been successfully induced between generative cells of the same species mentioned above,between generative cells of Z.Candida and L.radiata, between generative cells and petal protoplasts in L.radiata,and between generative cells of L.radiata and hypocotyl protoplasts of Brassica napus.In all cases either homokaryons or heterokaryons could be obtained.Fusion of nuclei was observed sometimes in homokaryons of generative cells in L.radiata.The generative nuclei in fusion products could be well identified by labelling the generative cells before fusion with DAPI.FDA test demonstrated that most of the fusion products were viable.Factors affecting fusion efficiency including cell density, PEG concentration,duration of PEG treatment and effect of calcium ions were studied in fusion of generative cells in Z.Candida.Our experiments indicate that isolated generative cells are likely to be deprived of cell walls and may be regarded as a special kind of protoplasts for direct fusion experiments.
基金
863高技术
高等学校博士点专项基金资助的项目
关键词
生殖细胞
花粉
原生质体
融合
Generative cell
Protoplast
Fusion
Zephyranthes Candida
Lycoris ra-diata
