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Purine biosynthetic enzyme ATase2 is involved in the regulation of early chloroplast development and chloroplast gene expression in Arabidopsis

机译:嘌呤生物合成酶ATase2参与拟南芥中早期叶绿体发育和叶绿体基因表达的调控

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摘要

To investigate the molecular mechanism of chloroplast biogenesis and development, we characterized an Arabidopsis mutant (dg169, delayed greening 169) which showed growth retardation and delayed greening phenotype in leaves. Newly emerged chlorotic leaves recovered gradually with leaf development in the mutant, and the mature leaves showed similar phenotype to those of wild-typewild-type plants. Compared with wild-type, the chloroplasts were oval-shaped and smaller and the thylakoid membranes were less abundant in yellow section of young leaves of dg169. In addition, the functions of photosystem II (PSII) and photosystem I (PSI) were also impaired. Furthermore, the amount of core subunits of PSII and PSI, as well as PSII and PSI complexes reduced in yellow section of young leaves of dg169. Map-based positional cloning identified that phenotype of dg169 was attributed to a point mutation of ATase2 which converts the conserved Ile-155 residue to Asn. ATase2 catalyzes the first step of de novo purine biosynthesis. This mutation resulted in impaired purine synthesis and a significant decrease in ATP, ADP, GTP and GDP contents. The analysis of ATase2-GFP protein fusion showed that ATase2 was localized to nucleoid of chloroplasts. Our results further demonstrated that the levels of PEP-dependent transcripts in yellow section of young leaves of dg169 were decreased while NEP-dependent and both PEP- and NEP-dependent transcripts and chloroplast DNA replications were increased. The results in this study suggest that ATase2 plays an essential role in early chloroplast development through maintaining PEP function.
机译:为了研究叶绿体生物发生和发展的分子机制,我们表征了拟南芥突变体(dg169,延迟绿化169),该突变体在叶片中显示出生长迟缓和延迟绿化表型。随着突变体叶片的发育,新出现的绿藻叶片逐渐恢复,成熟叶片表现出与野生型野生植物相似的表型。与野生型相比,dg169幼叶的黄色部分叶绿体呈椭圆形且较小,类囊体膜较少。此外,光系统II(PSII)和光系统I(PSI)的功能也受到损害。此外,dg169幼叶的黄色部分中PSII和PSI核心亚基的数量以及PSII和PSI复合物的数量减少。基于图的位置克隆确定dg169的表型归因于ATase2的点突变,该突变将保守的Ile-155残基转化为Asn。 ATase2催化从头进行嘌呤生物合成的第一步。这种突变导致嘌呤合成受损,ATP,ADP,GTP和GDP含量显着下降。对ATase2-GFP蛋白融合物的分析表明,ATase2位于叶绿体的核苷中。我们的结果进一步证明,dg169幼叶黄色部分中PEP依赖性转录物的水平降低,而NEP依赖性以及PEP和NEP依赖性转录物和叶绿体DNA复制均增加。这项研究的结果表明,ATase2通过维持PEP功能在早期叶绿体发育中起重要作用。

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