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拟南芥根内皮层细胞中 SCARECROW 启动子驱动表达的细菌汞转运蛋白 MerC 增强了汞在地上部的积累。

SCARECROW promoter-driven expression of a bacterial mercury transporter MerC in root endodermal cells enhances mercury accumulation in Arabidopsis shoots.

机构信息

Department of Public Health, School of Pharmacy, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8641, Japan.

出版信息

Planta. 2019 Aug;250(2):667-674. doi: 10.1007/s00425-019-03186-3. Epub 2019 May 18.

Abstract

Mercury accumulation in Arabidopsis shoots is accelerated by endodermis specific expression of fusion proteins of a bacterial mercury transporter MerC and a plant SNARE SYP121 under control of SCARECROW promoter. We previously demonstrated that the CaMV 35S RNA promoter (p35S)-driven ubiquitous expression of a bacterial mercury transporter MerC, fused with SYP121, an Arabidopsis SNARE protein increases mercury accumulation of Arabidopsis. To establish an improved fine-tuned mercury transport system in plants for phytoremediation, the present study generated and characterized transgenic Arabidopsis plants expressing MerC-SYP121 specifically in the root endodermis, which is a crucial cell type for root element uptake. We generated four independent transgenic Arabidopsis lines expressing a transgene encoding mCherry-MerC-SYP121 under the control of the endodermis-specific SCARECROW promoter (hereafter pSCR lines). Quantitative real-time PCR analysis showed that expression levels of the transgene in roots of the pSCR lines were 3-23% of the p35S driven-overexpressing line. Confocal microscopy analysis showed that mCherry-MerC-SYP121 was dominantly expressed in the endodermis of the meristematic zone as well as in the mature zone of the pSCR roots. Mercury accumulation in shoots of the pSCR lines exposed to inorganic mercury was overall higher than the wild-type and comparable to the p35S over-expressing line. These results suggest that endodermis-specific expression of the MerC-SYP121 fusion proteins in plant roots sufficiently enhances mercury uptake and accumulation into shoots, which would be an ideal phenotype for phytoremediation of mercury-contaminated environments.

摘要

在 SCARECROW 启动子的控制下,通过将细菌汞转运蛋白 MerC 和植物 SNARE SYP121 的融合蛋白在根内皮细胞中特异性表达,加速拟南芥地上部分的汞积累。我们之前证明,在 CaMV 35S RNA 启动子(p35S)驱动下,细菌汞转运蛋白 MerC 与 Arabidopsis SNARE 蛋白 SYP121 的融合体的普遍表达增加了拟南芥的汞积累。为了在植物中建立一个改进的、微调的汞转运系统用于植物修复,本研究生成并表征了在根内皮细胞中特异性表达 MerC-SYP121 的转基因拟南芥植物,根内皮细胞是根元素吸收的关键细胞类型。我们生成了四个独立的转基因拟南芥系,在根内皮细胞特异性 SCARECROW 启动子(以下简称 pSCR 系)的控制下表达编码 mCherry-MerC-SYP121 的转基因。定量实时 PCR 分析显示,pSCR 系中根中转基因的表达水平是 p35S 驱动过表达系的 3-23%。共焦显微镜分析显示,mCherry-MerC-SYP121 在分生区和 pSCR 根的成熟区的内皮细胞中主要表达。暴露于无机汞的 pSCR 系地上部分的汞积累总体上高于野生型,与 p35S 过表达系相当。这些结果表明,植物根系中 MerC-SYP121 融合蛋白的内皮细胞特异性表达足以增强汞的吸收和向地上部分的积累,这将是汞污染环境植物修复的理想表型。

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