Pawlowski W P, Torbert K A, Rines H W, Somers D A
Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul 55108, USA.
Plant Mol Biol. 1998 Nov 1;38(4):597-607. doi: 10.1023/a:1006090731414.
An irregular pattern of transgene silencing was revealed in expression and inheritance studies conducted over multiple generations following transgene introduction by microprojectile bombardment of allohexaploid cultivated oat (Avena sativa L.). Expression of two transgenes, bar and uidA, delivered on the same plasmid was investigated in 23 transgenic oat lines. Twenty-one transgenic lines, each derived from an independently selected transformed tissue culture, showed expression of both bar and uidA while two lines expressed only bar. The relationship of the transgenic phenotypes to the presence of the transgenes in the study was determined using (1) phenotypic scoring combined with Southern blot analyses of progeny, (2) coexpression of the two transgenic phenotypes since the two transgenes always cosegregated, and (3) reactivation of a transgenic phenotype in self-pollinated progenies of transgenic plants that did not exhibit a transgenic phenotype. Transgene silencing was observed in 19 of the 23 transgenic lines and resulted in distorted segregation of transgenic phenotypes in 10 lines. Silencing and inheritance distortions were irregular and unpredictable. They were often reversible in a subsequent generation of self-pollinated progeny and abnormally segregating progenies were as likely to trace back to parents that exhibited normal segregation in a previous generation as to parents showing segregation distortions. Possible causes of the irregular patterns of transgene silencing are discussed.
在用微粒轰击法将转基因导入异源六倍体栽培燕麦(Avena sativa L.)后,通过多代进行的表达和遗传研究揭示了一种不规则的转基因沉默模式。在23个转基因燕麦品系中研究了位于同一质粒上的两个转基因bar和uidA的表达情况。21个转基因品系,每个品系都源自一个独立选择的转化组织培养物,同时显示出bar和uidA的表达,而两个品系仅表达bar。研究中使用以下方法确定转基因表型与转基因存在之间的关系:(1)对后代进行表型评分并结合Southern印迹分析;(2)由于两个转基因总是共分离,因此对两种转基因表型进行共表达分析;(3)在未表现出转基因表型的转基因植物的自花授粉后代中重新激活转基因表型。在23个转基因品系中的19个中观察到了转基因沉默,导致10个品系中的转基因表型分离出现扭曲。沉默和遗传分离扭曲是不规则且不可预测的。它们在随后的自花授粉后代中通常是可逆的,并且异常分离的后代追溯到上一代表现出正常分离的亲本的可能性与追溯到表现出分离扭曲的亲本的可能性一样大。文中讨论了转基因沉默不规则模式的可能原因。