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黄瓜中硅转运蛋白基因CsLsi1的分离与功能鉴定

Isolation and functional characterization of CsLsi1, a silicon transporter gene in Cucumis sativus.

作者信息

Sun Hao, Guo Jia, Duan Yaoke, Zhang Tiantian, Huo Heqiang, Gong Haijun

机构信息

College of Horticulture, Northwest A&F University, Yangling 712100, China.

Mid-Florida Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, Apopka, FL 32703, USA.

出版信息

Physiol Plant. 2017 Feb;159(2):201-214. doi: 10.1111/ppl.12515. Epub 2016 Nov 4.

Abstract

Cucumber (Cucumis sativus) is a widely grown cucurbitaceous vegetable that exhibits a relatively high capacity for silicon (Si) accumulation, but the molecular mechanism for silicon uptake remains to be clarified. Here we isolated and characterized CsLsi1, a gene encoding a silicon transporter in cucumber (cv. Mch-4). CsLsi1 shares 55.70 and 90.63% homology with the Lsi1s of a monocot and dicot, rice (Oryza sativa) and pumpkin (Cucurbita moschata), respectively. CsLsi1 was predominantly expressed in the roots, and application of exogenous silicon suppressed its expression. Transient expression in cucumber protoplasts showed that CsLsi1 was localized in the plasma membrane. Heterologous expression in Xenopus laevis oocytes showed that CsLsi1 evidenced influx transport activity for silicon but not urea or glycerol. Expression of cucumber CsLsi1-mGFP under its own promoter showed that CsLsi1 was localized at the distal side of the endodermis and the cortical cells in the root tips as well as in the root hairs near the root tips. Heterologous expression of CsLsi1 in a rice mutant defective in silicon uptake and the over-expression of this gene in cucumber further confirmed the role of CsLsi1 in silicon uptake. Our results suggest that CsLsi1 is a silicon influx transporter in cucumber. The cellular localization of CsLsi1 in cucumber roots is different from that in other plants, implying the possible effect of transporter localization on silicon uptake capability.

摘要

黄瓜(Cucumis sativus)是一种广泛种植的葫芦科蔬菜,具有相对较高的硅(Si)积累能力,但硅吸收的分子机制仍有待阐明。在此,我们分离并鉴定了CsLsi1,这是一个在黄瓜(品种Mch - 4)中编码硅转运蛋白的基因。CsLsi1与单子叶植物水稻(Oryza sativa)和双子叶植物南瓜(Cucurbita moschata)的Lsi1分别具有55.70%和90.63%的同源性。CsLsi1主要在根中表达,外源硅的施用会抑制其表达。在黄瓜原生质体中的瞬时表达表明CsLsi1定位于质膜。在非洲爪蟾卵母细胞中的异源表达表明CsLsi1具有硅的流入转运活性,但对尿素或甘油没有转运活性。在其自身启动子下黄瓜CsLsi1 - mGFP的表达表明,CsLsi1定位于根尖内皮层和皮层细胞的远端以及根尖附近的根毛中。CsLsi1在硅吸收缺陷的水稻突变体中的异源表达以及该基因在黄瓜中的过表达进一步证实了CsLsi1在硅吸收中的作用。我们的结果表明CsLsi1是黄瓜中的一种硅流入转运蛋白。CsLsi1在黄瓜根中的细胞定位与其他植物不同,这意味着转运蛋白定位可能对硅吸收能力有影响。

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