Pascual María Belén, Jing Zhong Ping, Kirby Edward G, Cánovas Francisco M, Gallardo Fernando
Departamento de Biología Molecular y Bioquímica, Facultad de Ciencias, Universidad de Málaga, and Instituto Andaluz de Biotecnología, Campus de Teatinos, E-29071 Málaga, Spain.
Phytochemistry. 2008 Jan;69(2):382-9. doi: 10.1016/j.phytochem.2007.07.031. Epub 2007 Sep 20.
Glutamine synthetase (GS) is the main enzyme involved in ammonia assimilation in plants and is the target of phosphinothricin (PPT), an herbicide commonly used for weed control in agriculture. As a result of the inhibition of GS, PPT also blocks photorespiration, resulting in the depletion of leaf amino acid pools leading to the plant death. Hybrid transgenic poplar (Populus tremula x P. alba INRA clone 7171-B4) overexpressing cytosolic GS is characterized by enhanced vegetative growth [Gallardo, F., Fu, J., Cantón, F.R., García-Gutiérrez, A., Cánovas, F.M., Kirby, E.G., 1999. Expression of a conifer glutamine synthetase gene in transgenic poplar. Planta 210, 19-26; Fu, J., Sampalo, R., Gallardo, F., Cánovas, F.M., Kirby, E.G., 2003. Assembly of a cytosolic pine glutamine synthetase holoenzyme in leaves of transgenic poplar leads to enhanced vegetative growth in young plants. Plant Cell Environ. 26, 411-418; Jing, Z.P., Gallardo, F., Pascual, M.B., Sampalo, R., Romero, J., Torres de Navarra, A., Cánovas, F.M., 2004. Improved growth in a field trial of transgenic hybrid poplar overexpressing glutamine synthetase. New Phytol. 164, 137-145], increased photosynthetic and photorespiratory capacities [El-Khatib, R.T., Hamerlynck, E.P., Gallardo, F., Kirby, E.G., 2004. Transgenic poplar characterized by ectopic expression of a pine cytosolic glutamine synthetase gene exhibits enhanced tolerance to water stress. Tree Physiol. 24, 729-736], enhanced tolerance to water stress (El-Khatib et al., 2004), and enhanced nitrogen use efficiency [Man, H.-M., Boriel, R., El-Khatib, R.T., Kirby, E.G., 2005. Characterization of transgenic poplar with ectopic expression of pine cytosolic glutamine synthetase under conditions of varying nitrogen availability. New Phytol. 167, 31-39]. In vitro plantlets of GS transgenic poplar exhibited enhanced resistance to PPT when compared with non-transgenic controls. After 30 days exposure to PPT at an equivalent dose of 275 g ha(-1), growth of GS transgenic poplar plantlets was 5-fold greater than controls. The response of young leaves to PPT treatment depends on physiological state as indicated by GS and Rubisco (LSU) levels. Young leaves from control plants, typically in a low differentiation state, respond to the herbicide showing up-regulation of GS and LSU. In contrast, young leaves from transgenic lines, with higher initial GS and LSU levels compared to control, display up-regulation of NADP(+)-isocitrate dehydrogenase. Differences between control and GS transgenics in their response to PPT are discussed in relation to their differences in photosynthetic and photorespiratory capacities (El-Khatib et al., 2004).
谷氨酰胺合成酶(GS)是植物中参与氨同化的主要酶,也是草丁膦(PPT)的作用靶点,草丁膦是农业中常用的一种除草剂。由于GS受到抑制,PPT还会阻断光呼吸,导致叶片氨基酸库耗尽,从而致使植物死亡。过量表达胞质GS的杂交转基因杨树(欧洲山杨×银白杨INRA克隆7171 - B4)具有营养生长增强的特征[Gallardo, F., Fu, J., Cantón, F.R., García - Gutiérrez, A., Cánovas, F.M., Kirby, E.G., 1999. 针叶树谷氨酰胺合成酶基因在转基因杨树中的表达。植物 210, 19 - 26;Fu, J., Sampalo, R., Gallardo, F., Cánovas, F.M., Kirby, E.G., 2003. 胞质松树谷氨酰胺合成酶全酶在转基因杨树叶片中的组装导致幼龄植物营养生长增强。植物细胞环境。26, 411 - 418;Jing, Z.P., Gallardo, F., Pascual, M.B., Sampalo, R., Romero, J., Torres de Navarra, A., Cánovas, F.M., 2004. 过量表达谷氨酰胺合成酶的转基因杂交杨树田间试验中生长改善。新植物学家。164, 137 - 145],光合和光呼吸能力增强[El - Khatib, R.T., Hamerlynck, E.P., Gallardo, F., Kirby, E.G., 2004. 以松树胞质谷氨酰胺合成酶基因异位表达为特征的转基因杨树表现出对水分胁迫的耐受性增强。树木生理学。24, 729 - 736],对水分胁迫的耐受性增强(El - Khatib等人,2004),以及氮利用效率提高[Man, H.-M., Boriel, R., El - Khatib, R.T., Kirby, E.G., 2005. 在不同氮供应条件下异位表达松树胞质谷氨酰胺合成酶的转基因杨树的特性。新植物学家。167, 31 - 39]。与非转基因对照相比,GS转基因杨树的离体苗对PPT表现出更强的抗性。在以275 g ha(-1)的等效剂量暴露于PPT 30天后,GS转基因杨树苗的生长比对照大五倍。幼叶对PPT处理的反应取决于生理状态,并以GS和Rubisco(LSU)水平为指标。对照植物的幼叶通常处于低分化状态,对除草剂的反应表现为GS和LSU上调。相比之下,转基因品系的幼叶与对照相比初始GS和LSU水平较高,表现出NADP(+)-异柠檬酸脱氢酶上调。对照和GS转基因植物在对PPT反应上的差异与其光合和光呼吸能力差异相关(El - Khatib等人,2004)。