Takahashi Hideyuki, Takahara Kentaro, Hashida Shin-nosuke, Hirabayashi Takayuki, Fujimori Tamaki, Kawai-Yamada Maki, Yamaya Tomoyuki, Yanagisawa Shuichi, Uchimiya Hirofumi
Iwate Biotechnology Research Center, Kitakami, Iwate 024-0003, Japan.
Plant Physiol. 2009 Sep;151(1):100-13. doi: 10.1104/pp.109.140665. Epub 2009 Jul 8.
Nicotinamide nucleotides (NAD and NADP) are important cofactors in many metabolic processes in living organisms. In this study, we analyzed transgenic Arabidopsis (Arabidopsis thaliana) plants that overexpress NAD kinase2 (NADK2), an enzyme that catalyzes the synthesis of NADP from NAD in chloroplasts, to investigate the impacts of altering NADP level on plant metabolism. Metabolite profiling revealed that NADP(H) concentrations were proportional to NADK activity in NADK2 overexpressors and in the nadk2 mutant. Several metabolites associated with the Calvin cycle were also higher in the overexpressors, accompanied by an increase in overall Rubisco activity. Furthermore, enhanced NADP(H) production due to NADK2 overexpression increased nitrogen assimilation. Glutamine and glutamate concentrations, as well as some other amino acids, were higher in the overexpressors. These results indicate that overexpression of NADK2 either directly or indirectly stimulates carbon and nitrogen assimilation in Arabidopsis under restricted conditions. Importantly, since neither up-regulation nor down-regulation of NADK2 activity affected the sum amount of NAD and NADP or the redox state, the absolute level of NADP and/or the NADP/NAD ratio likely plays a key role in regulating plant metabolism.
烟酰胺核苷酸(NAD和NADP)是生物体许多代谢过程中的重要辅助因子。在本研究中,我们分析了过表达NAD激酶2(NADK2)的转基因拟南芥植株,NADK2是一种在叶绿体中催化由NAD合成NADP的酶,以研究改变NADP水平对植物代谢的影响。代谢物谱分析表明,在NADK2过表达植株和nadk2突变体中,NADP(H)浓度与NADK活性成正比。与卡尔文循环相关的几种代谢物在过表达植株中也更高,同时伴随着整体 Rubisco 活性的增加。此外,由于NADK2过表达导致的NADP(H)产量增加促进了氮同化。过表达植株中的谷氨酰胺和谷氨酸浓度以及其他一些氨基酸含量更高。这些结果表明,在受限条件下,NADK2的过表达直接或间接刺激了拟南芥中的碳和氮同化。重要的是,由于NADK2活性的上调或下调均不影响NAD和NADP的总量或氧化还原状态,NADP的绝对水平和/或NADP/NAD比率可能在调节植物代谢中起关键作用。