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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.

SUBMITTER: Sun H

PROVIDER: S-EPMC6153473 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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

Sun Hao H Duan Yaoke Y Qi Xiaocui X Zhang Liyang L Huo Heqiang H Gong Haijun H

Annals of botany 20180901 4

<h4>Background and aims</h4>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 ge ...[more]

PMID: 29905780

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Project description:BACKGROUND: Despite its extensive use as a nitrogen fertilizer, the role of urea as a directly accessible nitrogen source for crop plants is still poorly understood. So far, the physiological and molecular aspects of urea acquisition have been investigated only in few plant species highlighting the importance of a high-affinity transport system. With respect to maize, a worldwide-cultivated crop requiring high amounts of nitrogen fertilizer, the mechanisms involved in the transport of urea have not yet been identified. The aim of the present work was to characterize the high-affinity urea transport system in maize roots and to identify the high affinity urea transporter. RESULTS: Kinetic characterization of urea uptake (<300 ?M) demonstrated the presence in maize roots of a high-affinity and saturable transport system; this system is inducible by urea itself showing higher Vmax and Km upon induction. At molecular level, the ORF sequence coding for the urea transporter, ZmDUR3, was isolated and functionally characterized using different heterologous systems: a dur3 yeast mutant strain, tobacco protoplasts and a dur3 Arabidopsis mutant. The expression of the isolated sequence, ZmDUR3-ORF, in dur3 yeast mutant demonstrated the ability of the encoded protein to mediate urea uptake into cells. The subcellular targeting of DUR3/GFP fusion proteins in tobacco protoplasts gave results comparable to the localization of the orthologous transporters of Arabidopsis and rice, suggesting a partial localization at the plasma membrane. Moreover, the overexpression of ZmDUR3 in the atdur3-3 Arabidopsis mutant showed to complement the phenotype, since different ZmDUR3-overexpressing lines showed either comparable or enhanced 15[N]-urea influx than wild-type plants. These data provide a clear evidence in planta for a role of ZmDUR3 in urea acquisition from an extra-radical solution. CONCLUSIONS: This work highlights the capability of maize plants to take up urea via an inducible and high-affinity transport system. ZmDUR3 is a high-affinity urea transporter mediating the uptake of this molecule into roots. Data may provide a key to better understand the mechanisms involved in urea acquisition and contribute to deepen the knowledge on the overall nitrogen-use efficiency in crop plants.

Dataset Information

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

Publications

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

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