Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET-UNR), Ocampo y Esmeralda, Predio CCT, Rosario 2000, Argentina.
NUI Galway, Plant Systems Biology Lab, Plant and AgriBiosciences Research Centre, Botany and Plant Science, Galway, Ireland.
J Exp Bot. 2015 Jun;66(11):3381-9. doi: 10.1093/jxb/erv150. Epub 2015 Apr 15.
Glutamate (Glu) is a taste enhancer that contributes to the characteristic flavour of foods. In fruit of tomato (Solanum lycopersicum L.), the Glu content increases dramatically during the ripening process, becoming the most abundant free amino acid when the fruit become red. There is also a concomitant increase in NADH-dependent glutamate dehydrogenase (GDH) activity during the ripening transition. This enzyme is located in the mitochondria and catalyses the reversible amination of 2-oxoglutarate to Glu. To investigate the potential effect of GDH on Glu metabolism, the abundance of GDH was altered by artificial microRNA technology. Efficient silencing of all the endogenous SlGDH genes was achieved, leading to a dramatic decrease in total GDH activity. This decrease in GDH activity did not lead to any clear morphological or metabolic phenotype in leaves or green fruit. However, red fruit on the transgenic plants showed markedly reduced levels of Glu and a large increase in aspartate, glucose and fructose content in comparison to wild-type fruit. These results suggest that GDH is involved in the synthesis of Glu in tomato fruit during the ripening processes. This contrasts with the biological role ascribed to GDH in many other tissues and species. Overall, these findings suggest that GDH has a major effect on the control of metabolic composition during tomato fruit ripening, but not at other stages of development.
谷氨酸(Glu)是一种味觉增强剂,有助于食物呈现独特的风味。在番茄(Solanum lycopersicum L.)果实中,Glu 含量在成熟过程中会急剧增加,当果实变红时,它成为最丰富的游离氨基酸。在成熟过程中,NADH 依赖性谷氨酸脱氢酶(GDH)的活性也会同时增加。这种酶位于线粒体中,催化 2-酮戊二酸到 Glu 的可逆氨基化反应。为了研究 GDH 对 Glu 代谢的潜在影响,我们利用人工 microRNA 技术改变 GDH 的丰度。成功实现了所有内源性 SlGDH 基因的有效沉默,导致总 GDH 活性显著下降。这种 GDH 活性的降低在叶片或绿色果实中并没有导致任何明显的形态或代谢表型。然而,与野生型果实相比,转基植物上的红色果实中 Glu 水平明显降低,天冬氨酸、葡萄糖和果糖含量显著增加。这些结果表明,GDH 参与了番茄果实成熟过程中 Glu 的合成。这与 GDH 在许多其他组织和物种中所具有的生物学功能形成对比。总的来说,这些发现表明,在番茄果实成熟过程中,GDH 对代谢组成的控制有重大影响,但在发育的其他阶段没有影响。