Identification and functional characterization of grapevine transporters that mediate glucose-6-phosphate uptake into plastids

Two grapevine glucose-6-Pi plastidial transporters differently expressed in plant organs and in response to environmental and hormonal signals are characterized. They are involved in starch accumulation in berries and canes.In grapevine, starch accumulation in the trunk is important for winter storage of carbon and in the flower for reproductive development. Berries also accumulate starch in their plastids, which are also involved in the synthesis of aroma compounds important for fruit quality. The present work characterizes two glucose-phosphate translocators (VvGPT1, VvGPT2) that control the accumulation of starch in grape amyloplasts. Three different splicing variants identified for VvGPT2 (VvGPT2 alpha, VvGPT2 beta and VvGPT2a"broken vertical bar) were more expressed in the leaves than in other organs. In contrast, VvGPT1 transcripts were more abundant in mature berries, canes and flowers than in the leaves. Expression of 35S-VvGPT1-GFP and 35S-VvGPT2a"broken vertical bar-GFP in tobacco leaf epidermal cells showed that the fusion proteins localized at the plastidial envelope. Complementation of the Arabidopsis pgi1-1 mutant impaired in leaf starch synthesis restored its ability to synthesize starch, demonstrating that VvGPT1 and VvGPT2a"broken vertical bar mediate the transport of glucose-6-Pi across the plastidial envelope. In grape cell suspensions, ABA, light and galactinol, together with sucrose and fructose, significantly increased the transcript abundance of VvGPT1, whereas VvGPT2a"broken vertical bar expression was affected only by sucrose. In addition, elicitation with methyl jasmonate strongly upregulated VvGPT1, VvGPT2a"broken vertical bar and VvPAL1, suggesting a role for GPTs in the production of secondary compounds in grapevine. Moreover, in grapevines cultivated in field conditions, VvGPT1 expression was higher in berries more exposed to the sun and subjected to higher temperatures. Although both VvGPT1 and VvGPT2 mediate the same function at the molecular level, they exhibit different expressioWe would like to thank Dr. Anja Schneider from the Ludwig-Maximilians, University of Munich for kindly providing Arabidopsis pgi1-1 seed. This work is supported by European Union Funds (FEDER/COMPETE Operational Competitiveness Programme) and Portuguese national Funds (FCT-Portuguese Foundation for Science and Technology): KBBE-2012-6-3117 "Inovinne", FCOMP-01-0124-FEDER-022692 and PTDC/AGR-ALI/100636/2008. HN was supported by a PhD grant from FCT (SFRH/BD/74257/2010). This work also benefited from the networking activities within the European funded COST ACTION FA1106 "QualityFruit".info:eu-repo/semantics/publishedVersio