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整合代谢物和基因表达谱揭示拟南芥中光敏色素A对多胺生物合成的调控。

Integrated metabolite and gene expression profiling revealing phytochrome A regulation of polyamine biosynthesis of Arabidopsis thaliana.

作者信息

Jumtee Kanokwan, Bamba Takeshi, Okazawa Atsushi, Fukusaki Eiichiro, Kobayashi Akio

机构信息

Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan.

出版信息

J Exp Bot. 2008;59(6):1187-200. doi: 10.1093/jxb/ern026. Epub 2008 Mar 28.

Abstract

In this study, metabolite profiling was demonstrated as a useful tool to plot a specific metabolic pathway, which is regulated by phytochrome A (phyA). Etiolated Arabidopsis wild-type (WT) and phyA mutant seedlings were irradiated with either far-red light (FR) or white light (W). Primary metabolites of the irradiated seedlings were profiled by gas chromatography time-of-flight mass spectrometry (GC/TOF-MS) to obtain new insights on phyA-regulated metabolic pathways. Comparison of metabolite profiles in phyA and WT seedlings grown under FR revealed a number of metabolites that contribute to the differences between phyA and the WT. Several metabolites, including some amino acids, organic acids, and major sugars, as well as putrescine, were found in smaller amounts in WT compared with the content in phyA seedlings grown under FR. There were also significant differences between metabolite profiles of WT and phyA seedlings during de-etiolation under W. The polyamine biosynthetic pathway was investigated further, because putrescine, one of the polyamines existing in a wide variety of living organisms, was found to be present in lower amounts in WT than in phyA under both light conditions. The expression levels of polyamine biosynthesis-related genes were investigated by quantitative real-time RT-PCR. The gene expression profiles revealed that the arginine decarboxylase 2 (ADC2) gene was transcribed less in the WT than in phyA seedlings under both light conditions. This finding suggests that ADC2 is negatively regulated by phyA during photomorphogenesis. In addition, S-adenosylmethionine decarboxylase 2 and 4 (SAMDC2 and SAMDC4) were found to be regulated by phyA but in a different manner from the regulation of ADC2.

摘要

在本研究中,代谢物谱分析被证明是描绘特定代谢途径的有用工具,该代谢途径受光敏色素A(phyA)调控。将黄化的拟南芥野生型(WT)和phyA突变体幼苗用远红光(FR)或白光(W)照射。通过气相色谱-飞行时间质谱(GC/TOF-MS)对照射后幼苗的初级代谢物进行谱分析,以获得关于phyA调控的代谢途径的新见解。比较在FR下生长的phyA和WT幼苗的代谢物谱,发现了许多导致phyA与WT之间差异的代谢物。与在FR下生长的phyA幼苗中的含量相比,WT中包括一些氨基酸、有机酸、主要糖类以及腐胺在内的几种代谢物含量较少。在W下脱黄化过程中,WT和phyA幼苗的代谢物谱也存在显著差异。由于腐胺(广泛存在于各种生物体中的多胺之一)在两种光照条件下在WT中的含量均低于phyA,因此对多胺生物合成途径进行了进一步研究。通过定量实时RT-PCR研究了多胺生物合成相关基因的表达水平。基因表达谱显示,在两种光照条件下,精氨酸脱羧酶2(ADC2)基因在WT中的转录水平均低于phyA幼苗。这一发现表明,在光形态建成过程中,ADC2受phyA负调控。此外,还发现S-腺苷甲硫氨酸脱羧酶2和4(SAMDC2和SAMDC4)受phyA调控,但调控方式与ADC2不同。

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