Consequences of the expression of a bacterial glucokinase in potato tubers, both in combination with and independently of a yeast-derived invertase

A. Fernie, J. Riesmeier, Annette Martiny, S. Ramalingam, L. Willmitzer, R. Trethewey
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引用次数: 27

Abstract

The aim of this work was to further define the metabolic factors that regulate carbohydrate metabolism in potato (Solanum tuberosum L. cv. Desiree) tubers. We previously found that glycolysis is induced (and starch accumulation reduced) in transgenic tubers in which a yeast invertase and a glucokinase from Zymomonas mobilis were expressed in the cytosol, whereas potato tuber size is dramatically increased when invertase expression is targeted to the apoplast. In this study we describe the further characterisation of potato tubers expressing a yeast invertase in the apoplast. We also report the generation of two novel transgenic plants in which the Z. mobilis glucokinase gene is expressed tuber-specifically (either in the wild type or apoplastic invertase-expressing back- ground). We evaluated the influence that increasing the glucokinase activity, independent of invertase activity, had on the shift in carbon partitioning, and assessed if the hexoses produced by the apoplastic cleavage of sucrose could be brought into metabolism. We found that expression of glucokinase either in the wild type or in the apoplastic invertase-expressing background led to changes in the levels of glucose and glucose 6-phosphate. However, these changes had little effect on carbon partitioning or tuber size with respect to the parent line. We conclude that neither the accumulation nor the phosphorylation of glucose play a pivotal role in the regulation of metabolism or morphology in the potato tuber.
细菌葡萄糖激酶在马铃薯块茎中表达的结果,无论是联合还是独立于酵母衍生的转化酶
本研究旨在进一步确定马铃薯碳水化合物代谢的调控因子。拿破仑情史)块茎。我们之前发现,在细胞质中表达酵母转化酶和来自活动单胞菌的葡萄糖激酶的转基因块茎中,糖酵解被诱导(和淀粉积累减少),而当转化酶在外质体中表达时,马铃薯块茎的大小显着增加。在这项研究中,我们描述了马铃薯块茎在外质体中表达酵母转化酶的进一步特征。我们还报道了两种新的转基因植物的产生,在这些植物中,Z. mobilis葡萄糖激酶基因在块茎上特异性表达(无论是在野生型还是在胞外逆转录酶表达背景下)。我们评估了增加独立于转化酶活性的葡萄糖激酶活性对碳分配转变的影响,并评估了由蔗糖的外胞体分裂产生的己糖是否可以被带入代谢。我们发现,无论是在野生型还是在载脂蛋白逆转录酶表达背景下,葡萄糖激酶的表达都会导致葡萄糖和葡萄糖6-磷酸水平的变化。然而,这些变化对亲本的碳分配和块茎大小影响不大。我们得出结论,葡萄糖的积累和磷酸化在马铃薯块茎的代谢或形态调节中都不起关键作用。
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