Napoli C., Lemieux C., Jorgensen R.
DNA Plant Technology Corporation, 6701 San Pablo Avenue, Oakland, California 94608.
Plant Cell. 1990 Apr;2(4):279-289. doi: 10.1105/tpc.2.4.279.
We attempted to overexpress chalcone synthase (CHS) in pigmented petunia petals by introducing a chimeric petunia CHS gene. Unexpectedly, the introduced gene created a block in anthocyanin biosynthesis. Forty-two percent of plants with the introduced CHS gene produced totally white flowers and/or patterned flowers with white or pale nonclonal sectors on a wild-type pigmented background; none of hundreds of transgenic control plants exhibited such phenotypes. Progeny testing of one plant demonstrated that the novel color phenotype co-segregated with the introduced CHS gene; progeny without this gene were phenotypically wild type. The somatic and germinal stability of the novel color patterns was variable. RNase protection analysis of petal RNAs isolated from white flowers showed that, although the developmental timing of mRNA expression of the endogenous CHS gene was not altered, the level of the mRNA produced by this gene was reduced 50-fold from wild-type levels. Somatic reversion of plants with white flowers to phenotypically parental violet flowers was associated with a coordinate rise in the steady-state levels of the mRNAs produced by both the endogenous and the introduced CHS genes. Thus, in the altered white flowers, the expression of both genes was coordinately suppressed, indicating that expression of the introduced CHS gene was not alone sufficient for suppression of endogenous CHS transcript levels. The mechanism responsible for the reversible co-suppression of homologous genes in trans is unclear, but the erratic and reversible nature of this phenomenon suggests the possible involvement of methylation.
我们试图通过导入一个嵌合的矮牵牛查尔酮合酶(CHS)基因,来在有色矮牵牛花瓣中过表达CHS。出乎意料的是,导入的基因在花青素生物合成过程中造成了阻断。42%导入CHS基因的植株开出了完全白色的花和/或在野生型有色背景上带有白色或浅的非克隆区域的斑纹花;数百株转基因对照植株中没有一株表现出这种表型。对一株植株的后代检测表明,这种新的颜色表型与导入的CHS基因共分离;没有该基因的后代在表型上是野生型。这种新颜色模式的体细胞和生殖稳定性是可变的。对从白色花中分离的花瓣RNA进行核糖核酸酶保护分析表明,虽然内源CHS基因mRNA表达的发育时间没有改变,但该基因产生的mRNA水平比野生型水平降低了50倍。开白花的植株向表型上亲本的紫色花的体细胞回复与内源和导入的CHS基因产生的mRNA稳态水平的协同上升有关。因此,在改变后的白色花中,两个基因的表达都被协同抑制,这表明导入的CHS基因的表达本身不足以抑制内源CHS转录本水平。负责同源基因反式可逆共抑制的机制尚不清楚,但这种现象的不稳定和可逆性质表明可能涉及甲基化。