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The GDC1 gene encodes a novel ankyrin domain-containing protein that is essential for grana formation in arabidopsis

机译:GDC1基因编码一种新型的含锚蛋白结构域的蛋白,这对于拟南芥中的格兰娜形成至关重要

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In land-plant chloroplasts, the grana play multiple roles in photosynthesis, including the potential increase of photosynthetic capacity in light and enhancement of photochemical efficiency in shade. However, the molecular mechanisms of grana formation remain elusive. Here, we report a novel gene, Grana-Deficient Chloroplast1 (GDC1), required for chloroplast grana formation in Arabidopsis (Arabidopsis thaliana). In the chloroplast of knockout mutant gdc1-3, only stromal thylakoids were observed, and they could not stack together to form appressed grana. The mutant exhibited seedling lethality with pale green cotyledons and true leaves. Further blue native-polyacrylamide gel electrophoresis analysis indicated that the trimeric forms of Light-Harvesting Complex II (LHCII) were scarcely detected in gdc1-3, confirming previous reports that the LHCII trimer is essential for grana formation. The Lhcb1 protein, the major component of the LHCIIb trimer, was substantially reduced, and another LHCIIb trimer component, Lhcb2, was slightly reduced in the gdc1-3 mutant, although their transcription levels were not altered in the mutant. This suggests that defective LHCII trimer formation in gdc1-3 is due to low amounts of Lhcb1 and Lhcb2. GDC1 encodes a chloroplast protein with an ankyrin domain within the carboxyl terminus. It was highly expressed in Arabidopsis green tissues, and its expression was induced by photosignaling pathways. Immunoblot analysis of the GDC1- green fluorescent protein (GFP) fusion protein in 35S::GDC1-GFP transgenic plants with GFP antibody indicates that GDC1 is associated with an approximately 440-kD thylakoid protein complex instead of the LHCII trimer. This shows that GDC1 may play an indirect role in LHCII trimerization during grana formation.
机译:在陆地植物的叶绿体中,谷物在光合作用中发挥多种作用,包括潜在增加光合能力和增强光化学效率。但是,颗粒形成的分子机制仍然难以捉摸。在这里,我们报告了一个新的基因,缺乏拟南芥叶绿体1(GDC1),拟南芥(Arabidopsis thaliana)中的叶绿体颗粒形成需要。在敲除突变体gdc1-3的叶绿体中,仅观察到间质类囊体,它们不能堆叠在一起形成贴壁的杂种。该突变体表现出幼苗杀伤力,其子叶为浅绿色,叶片为真叶。进一步的蓝色天然聚丙烯酰胺凝胶电泳分析表明,在gdc1-3中几乎未检测到三聚体形式的光捕获复合体II(LHCII),这证实了先前的报道,LHCII三聚体对于形成颗粒是必不可少的。 Lhcb1蛋白是LHCIIb三聚体的主要成分,在gdc1-3突变体中被大量还原,而另一个LHCIIb三聚体成分Lhcb2在gdc1-3突变体中被略微还原,尽管它们的转录水平没有改变。这表明gdc1-3中LHCII三聚体的缺陷形成是由于Lhcb1和Lhcb2的含量较低。 GDC1编码在羧基末端具有锚蛋白结构域的叶绿体蛋白。它在拟南芥绿色组织中高表达,并且其表达是由光信号途径诱导的。带有GFP抗体的35S :: GDC1-GFP转基因植物中GDC1-绿色荧光蛋白(GFP)融合蛋白的免疫印迹分析表明,GDC1与大约440 kD类囊体蛋白复合物相关,而不是与LHCII三聚体相关。这表明GDC1可能在格兰娜形成过程中的LHCII三聚化中起间接作用。

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