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Activation of Sucrose Transport in Defoliated Lolium perenne L.: An Example of Apoplastic Phloem Loading Plasticity

机译:落叶的多年生黑麦草中蔗糖运输的活化:质外生韧皮部负载可塑性的一个例子

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The pathway of carbon phloem loading was examined in leaf tissues of the forage grass Lolium perenne. The effect of defoliation (leaf blade removal) on sucrose transport capacity was assessed in leaf sheaths as the major carbon source for regrowth. The pathway of carbon transport was assessed via a combination of electron microscopy, plasmolysis experiments and plasma membrane vesicles (PMVs) purified by aqueous two-phase partitioning from the microsomal fraction. Results support an apoplastic phloem loading mechanism. Imposition of an artificial proton-motive force to PMVs from leaf sheaths energized an active, transient and saturable uptake of sucrose (Suc). The affinity of Suc carriers for Suc was 580 μM in leaf sheaths of undefoliated plants. Defoliation induced a decrease of Km followed by an increase of Vmax. A transporter was isolated from stubble (including leaf sheaths) cDNA libraries and functionally expressed in yeast. The level of L.perenne SUcrose Transporter 1 (LpSUT1) expression increased in leaf sheaths in response to defoliation. Taken together, the results indicate that Suc transport capacity increased in leaf sheaths of L. perenne in response to leaf blade removal. This increase might imply de novo synthesis of Suc transporters, including LpSUT1, and may represent one of the mechanisms contributing to rapid refoliation.
机译:在饲草草黑麦草的叶子组织中检查了碳韧皮部负载的途径。评估了叶鞘作为再生的主要碳源时的脱叶(叶片的去除)对蔗糖运输能力的影响。通过电子显微镜,溶质实验和通过从微粒体级分中进行水相两相分配纯化的质膜囊泡(PMV)的组合来评估碳迁移的途径。结果支持外生韧皮部加载机制。从叶鞘向PMV施加人工质子动力可主动,短暂和饱和地吸收蔗糖(Suc)。在未落叶植物的叶鞘中,Suc载体对Suc的亲和力为580μM。落叶导致K m 减小,然后V max 增大。从残茬(包括叶鞘)cDNA文库中分离出转运蛋白,并在酵母中功能性表达。响应叶的脱落,紫苏糖蔗糖转运蛋白1(LpSUT1)的表达水平在叶鞘中增加。两者合计,结果表明响应叶片去除,紫苏叶鞘中的Suc转运能力增加。这种增加可能意味着包括LpSUT1在内的Suc转运蛋白的从头合成,并且可能代表了导致快速再叶化的机制之一。

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