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酵母亚砷酸盐外排系统ACR3的异源表达提高了拟南芥对砷胁迫的耐受性。

Heterologous expression of the yeast arsenite efflux system ACR3 improves Arabidopsis thaliana tolerance to arsenic stress.

作者信息

Ali Waqar, Isner Jean-Charles, Isayenkov Stanislav V, Liu Wenju, Zhao Fang-Jie, Maathuis Frans J M

机构信息

Biology Department, Area 9, University of York, York YO10 5DD, UK.

Department of Genomics and Molecular Biotechnology, Institute of Food Biotechnology and Genomics, Osipovskogo str. 2a, Kiev-123, 04123, Ukraine.

出版信息

New Phytol. 2012 May;194(3):716-723. doi: 10.1111/j.1469-8137.2012.04092.x. Epub 2012 Mar 1.

Abstract

• Arsenic contamination has a negative impact on crop cultivation and on human health. As yet, no proteins have been identified in plants that mediate the extrusion of arsenic. Here, we heterologously expressed the yeast (Saccharomyces cerevisiae) arsenite efflux transporter ACR3 into Arabidopsis to evaluate how this affects plant tolerance and tissue arsenic contents. • ACR3 was cloned from yeast and transformed into wild-type and nip7;1 Arabidopsis. Arsenic tolerance was determined at the cellular level using vitality stains in protoplasts, in intact seedlings grown on agar plates and in mature plants grown hydroponically. Arsenic efflux was measured from protoplasts and from intact plants, and arsenic levels were measured in roots and shoots of plants exposed to arsenate. • At the cellular level, all transgenic lines showed increased tolerance to arsenite and arsenate and a greater capacity for arsenate efflux. With intact plants, three of four stably transformed lines showed improved growth, whereas only transgenic lines in the wild-type background showed increased efflux of arsenite into the external medium. The presence of ACR3 hardly affected tissue arsenic levels, but increased arsenic translocation to the shoot. • Heterologous expression of yeast ACR3 endows plants with greater arsenic resistance, but does not lower significantly arsenic tissue levels.

摘要

• 砷污染对作物种植和人类健康都有负面影响。目前,尚未在植物中鉴定出介导砷外排的蛋白质。在此,我们将酵母(酿酒酵母)亚砷酸盐外排转运蛋白ACR3在拟南芥中进行异源表达,以评估其对植物耐受性和组织砷含量的影响。

• 从酵母中克隆ACR3并将其转化到野生型和nip7;1拟南芥中。使用原生质体活力染色法在细胞水平上测定砷耐受性,在琼脂平板上生长的完整幼苗和水培生长的成熟植株中也进行测定。测定原生质体和完整植株的砷外排,并测量暴露于砷酸盐的植物根和地上部分的砷含量。

• 在细胞水平上,所有转基因株系对亚砷酸盐和砷酸盐的耐受性均增强,且砷酸盐外排能力更强。对于完整植株,四个稳定转化株系中的三个生长得到改善,而只有野生型背景下的转基因株系显示亚砷酸盐向外部培养基的外排增加。ACR3的存在几乎不影响组织砷含量,但增加了砷向地上部分的转运。

• 酵母ACR3的异源表达赋予植物更强的抗砷能力,但并未显著降低砷的组织含量。

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