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首页> 外文期刊>Journal of Experimental Botany >Cloning, localization and expression analysis of vacuolar sugar transporters in the CAM plant Ananas comosus (pineapple)
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Cloning, localization and expression analysis of vacuolar sugar transporters in the CAM plant Ananas comosus (pineapple)

机译：CAM植物菠萝（菠萝）中液泡糖转运蛋白的克隆，定位和表达分析

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摘要

In photosynthetic tissues of the CAM plant pineapple (Ananas comosus), storage of soluble sugars in the central vacuole during the daytime and their remobilization at night is required to provide carbon skeletons for nocturnal CO2 fixation. However, soluble sugars produced photosynthetically must also be exported to support growth processes in heterotrophic tissues. To begin to address how vacuolar sugar storage and assimilate partitioning are regulated in A. comosus, degenerate PCR and cDNA library screening were used to clone three candidate sugar transporters from the leaves of this species. Subcellular localization of the three transporters was investigated via expression of YFP-fusion proteins in tobacco epidermal cells and their co-localization with subcellular markers by confocal microscopy. Using this strategy, a putative hexose transporter (AcMST1) and a putative inositol transporter (AcINT1) were identified that both localized to the tonoplast, whereas a putative sucrose transporter (AcSUT1) was found to localize to prevacuolar compartments. A cDNA (AcMST2) with high similarity to a recently characterized tonoplast hexose transporter in Arabidopsis was also identified from an A. comosus fruit EST database. Analyses of transcript abundance indicated that AcMST1 was more highly expressed in fruits compared to leaves of A. comosus, whilst transcripts of AcINT1, AcSUT1, and AcMST2 were more abundant in leaves. Transcript abundance of AcINT1, the putative inositol transporter, showed day–night changes comparable to those of other CAM-related transcripts described in Mesembryanthemum crystallinum. The results are discussed in terms of the role of vacuolar sugar transporters in regulating carbon flow during the diel cycle in CAM plants.
机译：在CAM植物菠萝（Ananas comosus）的光合组织中，白天将可溶性糖储存在中央液泡中，晚上将其固定，以为夜间固定CO 2 提供碳骨架。但是，光合作用产生的可溶性糖也必须出口以支持异养组织中的生长过程。为了开始解决如何在空肠曲霉中调节液泡糖的贮藏和同化物分配，简并PCR和cDNA文库筛选用于从该物种的叶片中克隆三个候选糖转运蛋白。通过YFP融合蛋白在烟草表皮细胞中的表达及其与亚细胞标记物的共聚焦显微镜共定位，研究了这三种转运蛋白的亚细胞定位。使用此策略，确定了假定的己糖转运蛋白（AcMST1）和假定的肌醇转运蛋白（AcINT1）均定位于液泡膜，而发现的蔗糖转运蛋白（AcSUT1）定位于前真空腔。 cDNA（AcMST2）与拟南芥中最近鉴定的液泡塑料己糖转运蛋白也具有很高的相似性，它也可以从玉米曲霉果实EST数据库中鉴定出来。转录本丰度分析表明，与果曲霉的叶片相比，AcMST1在果实中的表达更高，而AcINT1，AcSUT1和AcMST2的转录本在叶片中更为丰富。推定的肌醇转运蛋白AcINT1的转录本丰富度显示了昼夜变化，与结晶膜中描述的其他CAM相关转录本相当。就液泡糖转运蛋白在CAM植物的diel循环中调节碳流量方面的作用进行了讨论。

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