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利用爪蟾卵母细胞对硬粒小麦 Na+转运系统的功能鉴定揭示了两个 HKT1;4 转运蛋白的多样性。

Functional characterization in Xenopus oocytes of Na+ transport systems from durum wheat reveals diversity among two HKT1;4 transporters.

机构信息

Plant Protection and Improvement Laboratory, Center of Biotechnology of Sfax (CBS)/University of Sfax, B.P. '1177' 3018, Sfax, Tunisia.

出版信息

J Exp Bot. 2014 Jan;65(1):213-22. doi: 10.1093/jxb/ert361. Epub 2013 Nov 5.

Abstract

Plant tolerance to salinity constraint involves complex and integrated functions including control of Na(+) uptake, translocation, and compartmentalization. Several members of the high-affinity K(+) transporter (HKT) family, which comprises plasma-membrane transporters permeable to K(+) and Na(+) or to Na(+) only, have been shown to play major roles in plant Na(+) and K(+) homeostasis. Among them, HKT1;4 has been identified as corresponding to a quantitative trait locus (QTL) of salt tolerance in wheat but was not functionally characterized. Here, we isolated two HKT1;4-type cDNAs from a salt-tolerant durum wheat (Triticum turgidum L. subsp. durum) cultivar, Om Rabia3, and investigated the functional properties of the encoded transporters using a two-electrode voltage-clamp technique, after expression in Xenopus oocytes. Both transporters displayed high selectivity for Na(+), their permeability to other monovalent cations (K(+), Li(+), Cs(+), and Rb(+)) being ten times lower than that to Na(+). Both TdHKT1;4-1 and TdHKT1;4-2 transported Na(+) with low affinity, although the half-saturation of the conductance was observed at a Na(+) concentration four times lower in TdHKT1;4-1 than in TdHKT1;4-2. External K(+) did not inhibit Na(+) transport through these transporters. Quinine slightly inhibited TdHKT1;4-2 but not TdHKT1;4-1. Overall, these data identified TdHKT1;4 transporters as new Na(+)-selective transporters within the HKT family, displaying their own functional features. Furthermore, they showed that important differences in affinity exist among durum wheat HKT1;4 transporters. This suggests that the salt tolerance QTL involving HKT1;4 may be at least in part explained by functional variability among wheat HKT1;4-type transporters.

摘要

植物对盐胁迫的耐受性涉及包括控制 Na(+)摄取、转运和区隔化在内的复杂和综合功能。高亲和力 K(+)转运体 (HKT) 家族的几个成员,其组成质膜转运体对 K(+)和 Na(+)或仅对 Na(+)具有通透性,已被证明在植物 Na(+)和 K(+)稳态中发挥主要作用。其中,HKT1;4 已被确定为小麦耐盐性的数量性状位点 (QTL),但尚未对其功能进行表征。在这里,我们从耐盐性强的杜伦小麦(Triticum turgidum L. subsp. durum)品种 Om Rabia3 中分离出两个 HKT1;4 型 cDNA,并在非洲爪蟾卵母细胞中表达后,使用双电极电压钳技术研究了编码转运体的功能特性。两种转运体对 Na(+)具有高选择性,对其他单价阳离子(K(+)、Li(+)、Cs(+)和 Rb(+))的通透性比 Na(+)低 10 倍。尽管 TdHKT1;4-1 的电导半饱和浓度是 TdHKT1;4-2 的四分之一,但两种转运体均以低亲和力转运 Na(+)。外部 K(+) 不抑制这些转运体通过 Na(+)的运输。奎宁轻微抑制 TdHKT1;4-2,但不抑制 TdHKT1;4-1。总的来说,这些数据将 TdHKT1;4 转运体鉴定为 HKT 家族中新的 Na(+)-选择性转运体,具有其自身的功能特征。此外,它们表明,杜伦小麦 HKT1;4 转运体之间在亲和力上存在重要差异。这表明涉及 HKT1;4 的耐盐性 QTL 至少部分可以用小麦 HKT1;4 型转运体之间的功能变异性来解释。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cffc/3883290/ec6b37ca979b/exbotj_ert361_f0001.jpg

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