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Overexpression of malate dehydrogenase in transgenic tobacco leaves: enhanced malate synthesis and augmented Al-resistance

机译:苹果酸脱氢酶在转基因烟叶中的过表达:苹果酸合成增强和铝抗性增强

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Numerous studies with transgenic plants have demonstrated that overexpression of enzymes related to organic acid metabolism under the control of CaMV 35S promoter increased organic acid exudation and Al-resistance. The synthesis of organic acids requires a large carbon skeleton supply from leaf photosynthesis. Thus, we produced transgenic tobacco overexpressing cytosolic malate dehydrogenase (MDH) cDNA from Arabidopsis thaliana (amdh) and the MDH gene from Escherichia coli (emdh), respectively, under the control of a leaf-specific light-inducible promoter (Rubisco small subunit promoter, PrbcS) in the present study. Our data indicated that an increase (120–130%) in MDH-specific activity in leaves led to an increase in malate content in the transgenic tobacco leaves and roots as well as a significant increase in root malate exudation compared with the WT plants under the acidic (pH 4.5) conditions irrespective of 300 μM Al3+ stress absence or presence. After being exposed to 25 μM Al3+ in a hydroponic solution, the transgenic plants exhibited stronger Al-tolerance than WT plants and the degree of A1 tolerance in the transgenic plants corresponded with the amount of malate secretion. When grown in an Al-stress perlite medium, the transgenic tobacco lines showed better growth than the WT plants. The results suggested that overexpression of MDH driven by the PrbcS promoter in transgenic plant leaves enhanced malate synthesis and improved Al-resistance.
机译:对转基因植物的大量研究表明,在CaMV 35S启动子的控制下,与有机酸代谢相关的酶的过表达增加了有机酸的渗出和耐铝性。有机酸的合成需要叶片光合作用提供大量的碳骨架。因此,我们在叶片特异性光诱导启动子(Rubisco小亚基启动子)的控制下,分别从拟南芥(amdh)和大肠杆菌(emdh)生产了转基因烟草过表达胞质苹果酸脱氢酶(MDH)cDNA和大肠杆菌(emdh)的MDH基因。 (PrbcS))。我们的数据表明,与野生型植物相比,叶片中MDH比活性的增加(120-130%)导致转基因烟草叶片和根中苹果酸含量的增加,以及根中苹果酸的分泌显着增加。酸性(pH 4.5)条件下,无论是否存在300μMAl 3 + 胁迫。在水培溶液中暴露于25μMAl 3 + 后,转基因植物显示出比WT植物更强的耐铝性,并且转基因植物中A1耐性的程度与苹果酸的分泌量相对应。当在铝胁迫的珍珠岩培养基中生长时,转基因烟草品系显示出比野生型植物更好的生长。结果表明,由PrbcS启动子驱动的MDH在转基因植物叶片中的过表达增强了苹果酸的合成并改善了耐铝性。

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