Evaluation of metabolic alteration in transgenic rice overexpressing dihydroflavonol-4-reductase
Evaluation of metabolic alteration in transgenic rice overexpressing dihydroflavonol-4-reductase
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DOI：
10.1093/aob/mcl162
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发表时间：
2006-10-01
期刊：
影响因子：
4.2
通讯作者：
Uchimiya, Hirofumi
中科院分区：
文献类型：
--
作者：
Takahashi, Hideyuki;Hayashi, Mitsunori;Uchimiya, Hirofumi
中心点背景和目的先前的研究已经表明，过表达具有二氢黄酮醇-4-还原酶（DFR）活性的YK 1的转基因水稻植物表现出生物和非生物胁迫耐受性。通过傅里叶变换离子回旋质谱法，在这样的转基因植物中也显示了代谢物的高通量谱。在这项研究中，毛细管电泳质谱分析（CE/MS），以确定精确的代谢产物，如有机酸，氨基酸和sugars.center.方法使用CE/MS，我们分析了几个代谢产物的糖酵解，三羧酸（TCA）循环和戊糖磷酸途径。此外，糖和离子的浓度是quantified.center. Key Results在YK 1（DFR）过表达植物中，顺乌头酸、异柠檬酸和2-酮戊二酸的浓度在叶中较高，而果糖-1，6-二磷酸和甘油醛-3-磷酸的浓度在根中较低。在种子中，游离氨基酸和金属的量发生了变化，而种子中的糖保持不变。在YK 1愈伤组织中，3-观察到谷胱甘肽的倍数增加，而谷胱甘肽过氧化物酶和谷胱甘肽还原酶的活性同时增加。结论YK 1（DFR）的过表达与在整个植物中分析的几种代谢物的量的轻微变化有关，而谷胱甘肽衍生物在悬浮培养的细胞中显著增加。increased.center
center dot Background and Aims Previous studies have shown that transgenic rice plants overexpressing YK1, which possesses dihydroflavonol-4-reductase (DFR) activity, showed biotic and abiotic stress tolerance. High throughput profiles of metabolites have also been shown in such transgenic plants by Fourier transform ion cyclotron mass spectrometry. In this study, capillary electrophoresis mass spectrometry analysis (CE/MS) was employed to identify precise metabolites such as organic acids, amino acids and sugars.center dot Methods Using CE/MS, we analysed several metabolites of glycolysis, the tricarboxylic acid (TCA) cycle and the pentose phosphate pathway. In addition, the concentrations of sugars and ion were quantified.center dot Key Results In YK1 (DFR)-overexpressing plants, the concentrations of cis-aconitate, isocitrate and 2-oxoglutarate were higher in leaves, whereas those of fructose-1,6-bisphosphate and glyceraldehyde-3-phosphate were lower in roots. In seeds, the amounts of free amino acids and metals were altered, whereas sugars in seeds were kept constant. In YK1 calli, an approx. 3-fold increase in glutathione was observed, whereas the activities of glutathione peroxidase and glutathione reductase were concomitantly increased.center dot Conclusions The overexpression of YK1 (DFR) was associated with slight changes in the amounts of several metabolites analysed in whole plants, whilst glutathione derivatives were substantially increased in suspension-cultured cells.