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类结节素26内在膜蛋白OsNIP3;2参与水稻侧根对亚砷酸盐的吸收。

The Nodulin 26-like intrinsic membrane protein OsNIP3;2 is involved in arsenite uptake by lateral roots in rice.

作者信息

Chen Yi, Sun Sheng-Kai, Tang Zhong, Liu Guidong, Moore Katie L, Maathuis Frans J M, Miller Antony J, McGrath Steve P, Zhao Fang-Jie

机构信息

Department of Sustainable Soils and Grassland Systems, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK.

Department of Metabolic Biology, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK.

出版信息

J Exp Bot. 2017 May 17;68(11):3007-3016. doi: 10.1093/jxb/erx165.

Abstract

Previous studies have shown that the Nodulin 26-like intrinsic membrane protein (NIP) Lsi1 (OsNIP2;1) is involved in arsenite [As(III)] uptake in rice (Oryza sativa). However, the role of other rice NIPs in As(III) accumulation in planta remains unknown. In the present study, we investigated the role OsNIP3;2 in As(III) uptake in rice. When expressed in Xenopus laevis oocytes, OsNIP3;2 showed a high transport activity for As(III). Quantitative real-time RT-PCR showed that the expression of OsNIP3;2 was suppressed by 5 µM As(III), but enhanced by 20 and 100 µM As(III). Transgenic rice plants expressing OsNIP3;2pro-GUS showed that the gene was predominantly expressed in the lateral roots and the stele region of the primary roots. Transient expression of OsNIP3;2:GFP fusion protein in rice protoplasts showed that the protein was localized in the plasma membrane. Knockout of OsNIP3;2 significantly decreased As concentration in the roots, but had little effect on shoot As concentration. Synchrotron microfocus X-ray fluorescence showed decreased As accumulation in the stele of the lateral roots in the mutants compared with wild-type. Our results indicate that OsNIP3;2 is involved in As(III) uptake by lateral roots, but its contribution to As accumulation in the shoots is limited.

摘要

先前的研究表明,结节蛋白26样内在膜蛋白(NIP)Lsi1(OsNIP2;1)参与水稻(Oryza sativa)对亚砷酸盐[As(III)]的吸收。然而,其他水稻NIPs在植物体内As(III)积累中的作用仍不清楚。在本研究中,我们调查了OsNIP3;2在水稻As(III)吸收中的作用。当在非洲爪蟾卵母细胞中表达时,OsNIP3;2对As(III)表现出高转运活性。定量实时RT-PCR表明,5 μM As(III)抑制OsNIP3;2的表达,但20和100 μM As(III)增强其表达。表达OsNIP3;2pro-GUS的转基因水稻植株表明,该基因主要在侧根和初生根的中柱区域表达。OsNIP3;2:GFP融合蛋白在水稻原生质体中的瞬时表达表明,该蛋白定位于质膜。敲除OsNIP3;2显著降低了根中的As浓度,但对地上部As浓度影响不大。同步辐射微聚焦X射线荧光显示,与野生型相比,突变体侧根中柱的As积累减少。我们的结果表明,OsNIP3;2参与侧根对As(III)的吸收,但其对地上部As积累的贡献有限。

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