Isolation and functional characterization of CsLsi2, a cucumber silicon efflux transporter gene

Abstract

Background and Aims Silicon has been proven to exert beneficial effects on plant growth and stress tolerance, and silicon accumulation varies among different plant species. Cucumber (Cucumis sativus) is a widely used dicot model for silicon accumulation, but little is known about the molecular mechanism of its silicon uptake. Previously, we isolated and characterized CsLsi1, a silicon influx transporter gene from cucumber. In this study, we cloned a putative silicon efflux transporter gene, CsLsi2, from cucumber and investigated its role in silicon uptake. Methods The expression pattern, transport activity, and subcellular and cellular localizations of CsLsi2 were investigated. The transport activity of CsLsi2 was determined in Xenopus laevis oocytes. The subcelluar and cellular localizations were conducted by transient expression of fused 35S:CsLsi2-eGFP in onion epidermal cells and expression of ProCsLsi2:CsLsi2-mGFP in cucumber, respectively. Key Results CsLsi2 was mainly expressed in the roots. Expression of CsLsi2-eGFP fusion sequence in onion epidermis cells showed that CsLsi2 was localized at the plasma membrane. Transient expression in Xenopus laevis oocytes showed that CsLsi2 demonstrated efflux but no influx transport activity for silicon, and the transport was energy-dependent. Expression of CsLsi2-mGFP under its own promoter revealed that CsLsi2 was mainly expressed on endodermal cells, showing no polar distribution. In combination with our previous work on CsLsi1, a model for silicon uptake in cucumber roots is proposed. Conclusion The results suggest that CsLsi2 is a silicon efflux transporter gene in cucumber. The coordination of CsLsi1 and CsLsi2 mediates silicon uptake in cucumber roots. This study may help us understand the molecular mechanism for silicon uptake in cucumber, one of the few dicots with a relatively high capacity for silicon accumulation.