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拟南芥中两种质体ATP/ADP转运体的分子生理学分析

Molecular physiological analysis of the two plastidic ATP/ADP transporters from Arabidopsis.

作者信息

Reiser Jens, Linka Nicole, Lemke Lilia, Jeblick Wolfgang, Neuhaus H Ekkehard

机构信息

Pflanzenphysiologie, Fachbereich Biologie, Universität Kaiserslautern, D-67663 Kaiserslautern, Germany.

出版信息

Plant Physiol. 2004 Nov;136(3):3524-36. doi: 10.1104/pp.104.049502. Epub 2004 Oct 29.

Abstract

Arabidopsis (Arabidopsis thaliana) possesses two isoforms of plastidic ATP/ADP transporters (AtNTT1 and AtNTT2) exhibiting similar biochemical properties. To analyze the function of both isoforms on the molecular level, we examined the expression pattern of both genes by northern-blot analysis and promoter-beta-glucuronidase fusions. AtNTT1 represents a sugar-induced gene mainly expressed in stem and roots, whereas AtNTT2 is expressed in several Arabidopsis tissues with highest accumulation in developing roots and young cotyledons. Developing lipid-storing seeds hardly contained AtNTT1 or -2 transcripts. The absence of a functional AtNTT1 gene affected plant development only slightly, whereas AtNTT2T-DNA, AtNTT1-2T-DNA, and RNA interference (RNAi) plants showed retarded plant development, mainly characterized by a reduced ability to generate primary roots and a delayed chlorophyll accumulation in seedlings. Electron microscopic examination of chloroplast substructure also revealed an impaired formation of thylakoids in RNAi seedlings. Moreover, RNAi- and AtNTT1-2T-DNA plants showed reduced accumulation of the nuclear-encoded protein CP24 during deetiolation. Under short-day conditions reduced plastidic ATP import capacity correlates with a substantially reduced plant growth rate. This effect is absent under long-day conditions, strikingly indicating that nocturnal ATP import into chloroplasts is important. Plastidic ATP/ADP transport activity exerts significant control on lipid synthesis in developing Arabidopsis seeds. In total we made the surprising observation that plastidic ATP/ADP transport activity is not required to pass through the complete plant life cycle. However, plastidic ATP/ADP-transporter activity is required for both an undisturbed development of young tissues and a controlled cellular metabolism in mature leaves.

摘要

拟南芥(Arabidopsis thaliana)具有两种质体ATP/ADP转运体同工型(AtNTT1和AtNTT2),它们表现出相似的生化特性。为了在分子水平上分析这两种同工型的功能,我们通过Northern杂交分析和启动子-β-葡萄糖醛酸酶融合来检测这两个基因的表达模式。AtNTT1是一个糖诱导基因,主要在茎和根中表达,而AtNTT2在拟南芥的多个组织中表达,在发育中的根和幼嫩子叶中积累量最高。正在发育的储存脂质的种子几乎不含AtNTT1或-2转录本。功能性AtNTT1基因的缺失对植物发育影响较小,而AtNTT2 T-DNA、AtNTT1-2 T-DNA和RNA干扰(RNAi)植物显示出植物发育迟缓,主要表现为初生根生成能力降低和幼苗中叶绿素积累延迟。对叶绿体亚结构的电子显微镜检查还显示,RNAi幼苗中类囊体的形成受损。此外,RNAi和AtNTT1-2 T-DNA植物在脱黄化过程中核编码蛋白CP24的积累减少。在短日条件下,质体ATP输入能力降低与植物生长速率大幅降低相关。在长日条件下则不存在这种效应,这显著表明夜间ATP输入叶绿体很重要。质体ATP/ADP转运活性对拟南芥发育种子中的脂质合成有显著控制作用。总的来说,我们有一个惊人的发现,即质体ATP/ADP转运活性并非植物完整生命周期所必需。然而,质体ATP/ADP转运体活性对于幼嫩组织的正常发育和成熟叶片中受控的细胞代谢都是必需的。

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