Eigel L, Koop H U
Laboratory of Cell Biology and Cell Culture, Institute of Botany, University of Munich, FRG.
Mol Gen Genet. 1992 Jun;233(3):479-82. doi: 10.1007/BF00265447.
Defined numbers (1-5) of (donor) chloroplasts were transferred into (acceptor) protoplasts of plastid albino mutants by subprotoplasts/protoplast microfusion. Single transferred plastids gave rise to new organelle populations in the progeny of the fusion products when suitable combinations of plastomes were used or when selective pressure for the plastome transferred was applied. This process is termed "chloroplast cloning" and is the first reported case of "cloning" a cell organelle. The plastome combination and the presence or absence of selective pressure were found to influence the frequencies with which cell lines, containing both plastomes or acceptor or donor only, were obtained, and the number of cell generations needed for complete segregation - as measured by the duration of culture before the green donor plastome could be detected. The high frequency of cell lines and regenerated shoots recovered with donor plastome only, even when only a single chloroplast was transferred, leads to the conclusion that all organelles present in the fusion product contribute to the organelle population of the progeny, i.e. organelle death or loss are not regularly occurring events during plant regeneration from protoplasts in Nicotiana tabacum.
通过亚原生质体/原生质体微融合，将确定数量（1 - 5个）的（供体）叶绿体转移到质体白化突变体的（受体）原生质体中。当使用合适的质体基因组组合或对转移的质体基因组施加选择压力时，单个转移的质体在融合产物的后代中产生新的细胞器群体。这个过程被称为“叶绿体克隆”，是首次报道的“克隆”细胞器的案例。发现质体基因组组合以及选择压力的存在与否会影响获得同时含有两个质体基因组、仅含有受体或供体质体基因组的细胞系的频率，以及完全分离所需的细胞世代数——通过在检测到绿色供体质体基因组之前的培养持续时间来衡量。即使仅转移了单个叶绿体，仅含有供体质体基因组的细胞系和再生芽的高频率回收表明，融合产物中存在的所有细胞器都对后代的细胞器群体有贡献，即在烟草原生质体再生植物的过程中，细胞器死亡或丢失并非经常发生的事件。
