Flachmann R, Kühlbrandt W
European Molecular Biology Laboratory, Heidelberg, Germany.
Plant Cell. 1995 Feb;7(2):149-60. doi: 10.1105/tpc.7.2.149.
Transgenic tobacco plants expressing antisense RNA directed against the multigene family of the light-harvesting complex of photosystem II (LHCII) were raised and analyzed biochemically and physiologically. A partial 5' terminal sequence with 509 nucleotides complementary to cab (chlorophyll a/b binding protein) genes reduced the amount of transcript to almost undectectable levels. We demonstrated for endogenous genes that a 5' terminal sequence with only 52 to 105 nucleotides complementary to the transit sequence of cab can be equally efficient in gene repression. Chlorophyll content and chlorophyll a-to-chlorophyll b ratios of thylakoid membranes isolated from transgenic plants were unchanged in comparison with the wild type. Photosynthetic oxygen evolution and in vivo-measured chlorophyll fluorescence of the transformants showed that LHCII accumulates to normal levels. The reduced level of cab mRNA did not correlate with the amount of LHCII in thylakoids. This indicates that transcriptional regulation is not the rate-limiting step in the biogenesis of the LHCII apoprotein. The antenna size of photosystem II is therefore modulated by yet undiscovered posttranscriptional mechanisms.
培育出了表达针对光系统II捕光复合体(LHCII)多基因家族的反义RNA的转基因烟草植株,并对其进行了生化和生理分析。一个具有509个核苷酸、与cab(叶绿素a/b结合蛋白)基因互补的部分5'末端序列将转录本的量减少到几乎检测不到的水平。我们证明,对于内源基因,一个仅具有52至105个核苷酸、与cab转运序列互补的5'末端序列在基因抑制方面同样有效。与野生型相比,从转基因植物中分离出的类囊体膜的叶绿素含量和叶绿素a与叶绿素b的比率没有变化。转化体的光合放氧和体内测量的叶绿素荧光表明,LHCII积累到正常水平。cab mRNA水平的降低与类囊体中LHCII的量无关。这表明转录调控不是LHCII脱辅基蛋白生物合成中的限速步骤。因此,光系统II的天线大小是由尚未发现的转录后机制调节的。