Flachmann R
European Molecular Biology Laboratory, Heidelberg, Germany.
Plant Physiol. 1997 Mar;113(3):787-94. doi: 10.1104/pp.113.3.787.
Plants with genes coding for chlorophyll a/b-binding proteins of light-harvesting complex II (LHCII) in antisense orientation (Lhcb) that are characterized by severely reduced Lhcb transcript levels (below 10% of wild type) do not show a bleached phenotype due to a specific loss of the polypeptide. To produce such a phenotype, a conceptually different antisense approach was tested with a dual-functional transcript encoding the gene for hygromycin phosphotransferase and the transit sequence of Lhcb1-2 in the antisense orientation. Using increasing concentrations of hygromycin, transformants with Lhcb steady-state levels as low as 9% of wild type were regenerated and grown in a growth chamber. Together with Lhcb antisense plants obtained in an earlier study, these antisense plants were analyzed biochemically for their photosystem II (PSII) antenna composition under varying light conditions. All antisense plants showed a characteristic low-irradiance-induced increase of their PSII antenna size as determined by higher chlorophyll concentrations, an increased content of LHCII, and a constant chlorophyll b-to-lutein ratio in comparison with control plants. One to 5% of the total Lhcb transcript amount was sufficient to allow unrestricted formation of the PSII antenna at low irradiance, suggesting that LHCH biogenesis is not controlled primarily by transcription.
具有以反义方向编码光捕获复合物II(LHCII)的叶绿素a/b结合蛋白的基因(Lhcb)的植物,其特征在于Lhcb转录水平严重降低(低于野生型的10%),但并未因多肽的特异性缺失而表现出漂白表型。为了产生这种表型,测试了一种概念上不同的反义方法,即使用一种双功能转录本,其以反义方向编码潮霉素磷酸转移酶基因和Lhcb1-2的转运序列。使用浓度不断增加的潮霉素,再生出Lhcb稳态水平低至野生型9%的转化体,并在生长室中培养。与早期研究中获得的Lhcb反义植物一起,对这些反义植物在不同光照条件下的光系统II(PSII)天线组成进行了生化分析。与对照植物相比,所有反义植物在低辐照下均表现出PSII天线大小的特征性低辐照诱导增加,这由更高的叶绿素浓度、LHCII含量增加以及叶绿素b与叶黄素的恒定比例所确定。总Lhcb转录本量的1%至5%足以在低辐照下无限制地形成PSII天线,这表明LHCH的生物合成主要不是由转录控制的。