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过表达拟南芥 NAD 激酶基因的水稻植株的代谢组学和光化学分析。

Metabolome and photochemical analysis of rice plants overexpressing Arabidopsis NAD kinase gene.

机构信息

Institute of Molecular and Cellular Biosciences, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-0032, Japan.

出版信息

Plant Physiol. 2010 Apr;152(4):1863-73. doi: 10.1104/pp.110.153098. Epub 2010 Feb 12.

Abstract

The chloroplastic NAD kinase (NADK2) is reported to stimulate carbon and nitrogen assimilation in Arabidopsis (Arabidopsis thaliana), which is vulnerable to high light. Since rice (Oryza sativa) is a monocotyledonous plant that can adapt to high light, we studied the effects of NADK2 expression in rice by developing transgenic rice plants that constitutively expressed the Arabidopsis chloroplastic NADK gene (NK2 lines). NK2 lines showed enhanced activity of NADK and accumulation of the NADP(H) pool, while intermediates of NAD derivatives were unchanged. Comprehensive analysis of the primary metabolites in leaves using capillary electrophoresis mass spectrometry revealed elevated levels of amino acids and several sugar phosphates including ribose-1,5-bisphosphate, but no significant change in the levels of the other metabolites. Studies of chlorophyll fluorescence and gas change analyses demonstrated greater electron transport and CO2 assimilation rates in NK2 lines, compared to those in the control. Analysis of oxidative stress response indicated enhanced tolerance to oxidative stress in these transformants. The results suggest that NADP content plays a critical role in determining the photosynthetic electron transport rate in rice and that its enhancement leads to stimulation of photosynthesis metabolism and tolerance of oxidative damages.

摘要

质体 NAD 激酶(NADK2)被报道能刺激拟南芥(Arabidopsis thaliana)的碳氮同化,而拟南芥对高光很敏感。由于水稻(Oryza sativa)是一种能适应高光的单子叶植物,我们通过培育组成型表达拟南芥质体 NADK 基因(NK2 系)的转基因水稻植株来研究 NADK2 在水稻中的表达的影响。NK2 系表现出 NADK 活性增强和 NADP(H)库积累,而 NAD 衍生物的中间产物不变。使用毛细管电泳质谱对叶片中的初级代谢物进行综合分析表明,氨基酸和几种糖磷酸酯(包括核糖-1,5-二磷酸)的水平升高,但其他代谢物的水平没有明显变化。对叶绿素荧光和气体变化分析的研究表明,与对照相比,NK2 系中的电子传递和 CO2 同化速率更高。对氧化应激反应的分析表明,这些转化体对氧化应激的耐受性增强。结果表明,NADP 含量在决定水稻光合作用电子传递速率方面起着关键作用,其增强会刺激光合作用代谢并耐受氧化损伤。

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