Schaaf J., Walter M. H., Hess D.
Universitat Hohenheim, Institut fur Pflanzenphysiologie (260), D-70593 Stuttgart, Germany.
Plant Physiol. 1995 Jul;108(3):949-960. doi: 10.1104/pp.108.3.949.
NADP-dependent malic enzyme (NADP-ME, EC 1.1.1.40) catalyzes the oxidative decarboxylation of malate to pyruvate, producing CO2 and NADPH. We have examined regulatory properties of a 2.8-kb promoter-leader fragment of a bean (Phaseolus vulgaris L.) NADP-ME gene (PvME1) predicted to encode a cytosolic form of the enzyme by expression analysis of promoter-[beta]-glucuronidase fusions in transgenic tobacco plants. The PvME1 promoter directed strong expression in stems, which was confined to vascular and pith tissues, and was also active in floral and reproductive tissues. Wounding caused a marked induction of promoter activity, which was further strongly enhanced upon application of stimuli related to pathogen defense. Glutathione (reduced form) was the strongest inducer, but oxidized glutathione, fungal elicitor, cellulase, catalase, ascorbic acid, and NADPH were additional potent promoter-stimulating agents. Responsiveness to reduced glutathione was also shown at the level of PvME1 mRNA accumulation in bean plants. The putative contributions of NADP-ME gene expression to the plant defense response and possible mechanisms of defense gene regulation by conditions of oxidative stress as well as by H2O2 and antioxidant levels are discussed.
依赖于烟酰胺腺嘌呤二核苷酸磷酸(NADP)的苹果酸酶(NADP-ME,EC 1.1.1.40)催化苹果酸氧化脱羧生成丙酮酸,产生二氧化碳和还原型辅酶Ⅱ(NADPH)。我们通过在转基因烟草植株中对启动子-葡糖醛酸糖苷酶融合体进行表达分析,研究了菜豆(Phaseolus vulgaris L.)NADP-ME基因(PvME1)一个2.8 kb的启动子-前导片段的调控特性,该基因预计编码该酶的一种胞质形式。PvME1启动子在茎中指导强表达,这种表达局限于维管组织和髓组织,并且在花和生殖组织中也有活性。创伤引起启动子活性的显著诱导,在施加与病原体防御相关的刺激后,这种诱导进一步强烈增强。谷胱甘肽(还原型)是最强的诱导剂,但氧化型谷胱甘肽、真菌激发子、纤维素酶、过氧化氢酶、抗坏血酸和NADPH也是其他有效的启动子刺激剂。在菜豆植株中,PvME1 mRNA积累水平也显示出对还原型谷胱甘肽的响应性。讨论了NADP-ME基因表达对植物防御反应的推定贡献以及氧化应激条件以及过氧化氢和抗氧化剂水平对防御基因调控的可能机制。