Breia Richard, Conde Artur, Pimentel Diana, Conde Carlos, Fortes Ana Margarida, Granell Antonio, Gerós Hernâni
Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Braga, Portugal.
Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes e Alto Douro, Vila Real, Portugal.
Front Plant Sci. 2020 Jan 27;10:1753. doi: 10.3389/fpls.2019.01753. eCollection 2019.
The newly-identified SWEETs are high-capacity, low-affinity sugar transporters with important roles in numerous physiological mechanisms where sugar efflux is critical. SWEETs are desirable targets for manipulation by pathogens and their expression may be transcriptionally reprogrammed during infection. So far, few plant SWEET transporters have been functionally characterized, especially in grapevine. In this study, in the -susceptible variety "Trincadeira," we thoroughly analyzed modifications in the gene expression profile of key genes in -infected grape berries. VvSWEET7 and VvSWEET15 are likely to play an important role during fruit development and infection as they are strongly expressed at the green and mature stage, respectively, and were clearly up-regulated in response to infection. Also, infection down-regulated expression at the green stage, and expression at the veraison stage, and expression at the mature stage. VvSWEET7 was functionally characterized by heterologous expression in as a low-affinity, high-capacity glucose and sucrose transporter with a of 15.42 mM for glucose and a of 40.08 mM for sucrose. VvSWEET7-GFP and VvSWEET15-GFP fusion proteins were transiently expressed in epidermal cells and confocal microscopy allowed to observe that both proteins clearly localize to the plasma membrane. In sum, VvSWEETs transporters are important players in sugar mobilization during grape berry development and their expression is transcriptionally reprogrammed in response to infection.
新发现的SWEETs是高容量、低亲和力的糖转运蛋白,在许多糖外流至关重要的生理机制中发挥着重要作用。SWEETs是病原体操纵的理想靶标,其表达在感染过程中可能会发生转录重编程。到目前为止,很少有植物SWEET转运蛋白得到功能鉴定,尤其是在葡萄中。在本研究中,我们在易感品种“Trincadeira”中,深入分析了感染葡萄浆果中关键基因的基因表达谱变化。VvSWEET7和VvSWEET15可能在果实发育和感染过程中发挥重要作用,因为它们分别在绿色和成熟阶段强烈表达,并且在感染后明显上调。此外,感染下调了绿色阶段的表达、转色期的表达以及成熟阶段的表达。VvSWEET7通过在酵母中异源表达进行功能鉴定,它是一种低亲和力、高容量的葡萄糖和蔗糖转运蛋白,对葡萄糖的Km值为15.42 mM,对蔗糖的Km值为40.08 mM。VvSWEET7-GFP和VvSWEET15-GFP融合蛋白在烟草表皮细胞中瞬时表达,共聚焦显微镜观察发现这两种蛋白都明显定位于质膜。总之,VvSWEETs转运蛋白是葡萄浆果发育过程中糖转运的重要参与者,其表达在感染后发生转录重编程。