Institute of Plant Biology, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary.
Plant Cell Physiol. 2010 May;51(5):823-35. doi: 10.1093/pcp/pcq031. Epub 2010 Mar 15.
The crtB gene of Synechocystis sp. PCC 6803, encoding phytoene synthase, was inactivated in the Delta crtH mutant to generate a carotenoidless Delta crtH/B double mutant. Delta crtH mutant cells were used because they had better transformability than wild-type cells, most probably due to their adaptation to partial carotenoid deficiency. Cells of the Delta crtH/B mutant were light sensitive and could grow only under light-activated heterotrophic growth conditions in the presence of glucose. Carotenoid deficiency did not significantly affect the cellular content of phycobiliproteins while the chlorophyll content of the mutant cells decreased. The mutant cells exhibited no oxygen-evolving activity, suggesting the absence of photochemically active PSII complexes. This was confirmed by 2D electrophoresis of photosynthetic membrane complexes. Analyses identified only a small amount of a non-functional PSII core complex lacking CP43, while the monomeric and dimeric PSII core complexes were absent. On the other hand, carotenoid deficiency did not prevent formation of the cytochrome b(6)f complex and PSI, which predominantly accumulated in the monomeric form. Radioactive labeling revealed very limited synthesis of inner PSII antennae, CP47 and especially CP43. Thus, carotenoids are indispensable constituents of the photosynthetic apparatus, being essential not only for antioxidative protection but also for the efficient synthesis and accumulation of photosynthetic proteins and especially that of PSII antenna subunits.
将聚球藻 PCC 6803 的 crtB 基因(编码类胡萝卜素合酶)失活,以产生缺乏类胡萝卜素的 crtH/B 双突变体。选择 Delta crtH 突变体细胞是因为它们比野生型细胞具有更好的转化能力,这很可能是由于它们适应了部分类胡萝卜素缺乏。Delta crtH/B 突变体细胞对光敏感,只能在含有葡萄糖的光激活异养生长条件下生长。类胡萝卜素缺乏对藻胆蛋白的细胞含量没有显著影响,而突变体细胞的叶绿素含量下降。突变体细胞没有氧气产生活性,表明缺乏光化学活性 PSII 复合物。这通过光合膜复合物的二维电泳得到证实。分析仅鉴定出少量缺乏 CP43 的无功能 PSII 核心复合物,而单体和二聚 PSII 核心复合物不存在。另一方面,类胡萝卜素缺乏并不妨碍细胞色素 b(6)f 复合物和 PSI 的形成,它们主要以单体形式积累。放射性标记显示 PSII 天线、CP47 尤其是 CP43 的内天线的合成非常有限。因此,类胡萝卜素是光合作用装置的必需组成部分,不仅对抗氧化保护,而且对光合蛋白的有效合成和积累,尤其是 PSII 天线亚基的积累都是必不可少的。