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编码水稻类金属硫蛋白的rgMT基因在酿酒酵母和拟南芥中的表达。

Expression of the rgMT gene, encoding for a rice metallothionein-like protein in Saccharomyces cerevisiae and Arabidopsis thaliana.

作者信息

Jin Shumei, Sun Dan, Wang Ji, Li Ying, Wang Xinwang, Liu Shenkui

机构信息

Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field (SAVER),Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin 150040, People's Republic of China.

出版信息

J Genet. 2014 Dec;93(3):709-18. doi: 10.1007/s12041-014-0430-8.

Abstract

Metallothioneins (MTs) are cysteine-rich proteins of low molecular weight with many attributed functions, such as providing protection against metal toxicity, being involved in regulation of metal ions uptake that can impact plant physiology and providing protection against oxidative stress. However, the precise function of the metallothionein-like proteins such as the one coded for rgMT gene isolated from rice (Oryza sativa L.) is not completely understood. The whole genome analysis of rice (O. sativa) showed that the rgMT gene is homologue to the Os11g47809 on chromosome 11 of O. sativa sp. japonica genome. This study used the rgMT coding sequence to create transgenic lines to investigate the subcellular localization of the protein, as well as the impact of gene expression in yeast (Saccharomyces cerevisiae) and Arabidopsis thaliana under heavy metal ion, salt and oxidative stresses. The results indicate that the rgMT gene was expressed in the cytoplasm of transgenic cells. Yeast cells transgenic for rgMT showed vigorous growth compared to the nontransgenic controls when exposed to 7 mM CuCl2, 10 mM FeCl2, 1 M NaCl, 24 mM NaHCO3 and 3.2 mM H2O2, but there was no significant difference for other stresses tested. Similarly, Arabidopsis transgenic for rgMT displayed significantly improved seed germination rates over that of the control when the seeds were stressed with 100 μM CuCl2 or 1 mM H2O2. Increased biomass was observed in the presence of 100 μM CuCl2, 220 μM FeCl2, 3 mM Na2CO3, 5 mM NaHCO3 or 1 mM H2O2. These results indicate that the expression of the rice rgMT gene in transgenic yeast and Arabidopsis is implicated in improving their tolerance for certain salt and peroxide stressors.

摘要

金属硫蛋白(MTs)是一类富含半胱氨酸的低分子量蛋白质,具有多种功能,如提供对金属毒性的保护、参与影响植物生理的金属离子吸收调节以及提供对氧化应激的保护。然而,对于从水稻(Oryza sativa L.)中分离的rgMT基因编码的类金属硫蛋白等的精确功能尚未完全了解。水稻(O. sativa)的全基因组分析表明,rgMT基因与粳稻基因组第11号染色体上的Os11g47809同源。本研究利用rgMT编码序列创建转基因株系,以研究该蛋白质的亚细胞定位,以及在重金属离子、盐和氧化应激条件下在酵母(Saccharomyces cerevisiae)和拟南芥中基因表达的影响。结果表明,rgMT基因在转基因细胞的细胞质中表达。与非转基因对照相比,转rgMT基因的酵母细胞在暴露于7 mM CuCl2、10 mM FeCl2、1 M NaCl、24 mM NaHCO3和3.2 mM H2O2时显示出旺盛生长,但在测试的其他应激条件下没有显著差异。同样,当种子用100 μM CuCl2或1 mM H2O2处理时,转rgMT基因的拟南芥的种子发芽率比对照显著提高。在存在100 μM CuCl2、220 μM FeCl2、3 mM Na2CO3、5 mM NaHCO3或1 mM H2O2的情况下观察到生物量增加。这些结果表明,水稻rgMT基因在转基因酵母和拟南芥中的表达与提高它们对某些盐和过氧化物应激源的耐受性有关。

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