Eigel L, Oelmüller R, Koop H U
Botanical Institute, University of Munich, FRG.
Mol Gen Genet. 1991 Jul;227(3):446-51. doi: 10.1007/BF00273936.
A procedure is described by which it is possible to perform controlled microfusion of microscopically selected protoplast fusion partners with high efficiencies. The procedure is applied to fusion of Nicotiana tabacum (line 92V37. N. undulata cytoplasm) plastid albino protoplasts as a recipient and spontaneously formed subprotoplasts of green N. tabacum (line SR1) as donor. Products of individual electrofusion events are cloned via single cell nurse culture and the derived cell lines are analysed for the occurrence of variegated or green regenerating shoots, which are indicative of the establishment of the transferred organelles in the cell progeny. The plastid population in green regenerants recovered after the transfer of only two chloroplasts was demonstrated to have originated from the donor subprotoplast organelles by restriction analysis of total DNA using a plastome-specific probe.
本文描述了一种方法，通过该方法可以高效地对显微镜下选择的原生质体融合伙伴进行可控微融合。该方法应用于烟草（92V37品系，具波浪叶烟草细胞质）质体白化原生质体作为受体，与绿色烟草（SR1品系）自发形成的亚原生质体作为供体的融合。通过单细胞滋养培养对单个电融合事件的产物进行克隆，并对衍生的细胞系进行分析，以检测杂色或绿色再生芽的出现，这表明转移的细胞器在细胞后代中得以建立。通过使用质体基因组特异性探针进行总DNA的限制性分析，证明仅转移两个叶绿体后回收的绿色再生植株中的质体群体源自供体亚原生质体细胞器。
