Isolation and characterization of a metallothionein-1 protein in Chloris virgata Swartz that enhances stress tolerances to oxidative, salinity and carbonate stress in Saccharomyces cerevisiae
Isolation and characterization of a metallothionein-1 protein in Chloris virgata Swartz that enhances stress tolerances to oxidative, salinity and carbonate stress in Saccharomyces cerevisiae
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DOI：
10.1007/s10529-007-9396-4
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发表时间：
2007-05
影响因子：
2.7
通讯作者：
S. Nishiuchi;Shenkui Liu;T. Takano
中科院分区：
文献类型：
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作者：
S. Nishiuchi;Shenkui Liu;T. Takano
Chloris virgataSwartz (C. virgata) 是一种禾本科野生植物，发现于中国东北碱性土壤地区，对碳酸盐胁迫具有高度耐受性。我们构建了 C 的 cDNA 文库。用NaHCO3处理virgata幼苗，并从文库中分离出1型金属硫蛋白（MT1）基因（ChlMT1：AB294238）。 ChlMT1 的氨基酸序列包含 12 个半胱氨酸残基，构成 N 端和 C 端区域的 Cys-X-Cys（X = 除 Cys 之外的氨基酸）基序。 Northern 杂交表明，ChlMT1 的表达受到多种非生物胁迫的诱导，包括盐（NaCl 和 NaHCO3）、ROS 诱导剂（百草枯）和金属（CuSO4、ZnSO4 和 CoCl2）。叶子中的 ChlMT1 表达是由 200 mM NaCl 和 100 mM NaHCO3 诱导的。大约 5 μM 百草枯、500 μM Zn2+ 和 500 μM Co2+ 6 小时后也在叶片中诱导 ChlMT1 的表达，100 μM Cu2+ 24 小时后诱导其表达。 酿酒酵母在用 ChlMT1 基因转化时，对盐（NaCl 和 NaHCO3）和 ROS 的耐受性显着增加。
Chloris virgataSwartz (C. virgata) is a gramineous wild plant that is found in alkaline soil areas in northeast China and is highly tolerant to carbonate stress. We constructed a cDNA library fromC. virgataseedlings treated with NaHCO3, and isolated a type1 metallothionein (MT1) gene (ChlMT1: AB294238) from the library. The amino acid sequence of ChlMT1 contained 12 cysteine residues that constituted the Cys-X-Cys (X = amino acid except Cys) motifs in theN- andC-terminal regions. Northern hybridization showed that expression ofChlMT1was induced by several abiotic stresses, from salts (NaCl and NaHCO3), a ROS inducer (paraquat), and metals (CuSO4, ZnSO4, and CoCl2).ChlMT1expression in leaf was induced by 200 mM NaCl and 100 mM NaHCO3. About 5 μM Paraquat, 500 μM Zn2+, and 500 μM Co2+also induced expression ofChlMT1in leaf after 6 h, and 100 μM Cu2+induced it after 24 h.Saccharomyces cerevisiaewhen transformed with theChlMT1gene had dramatically increased tolerances to salts (NaCl and NaHCO3) and ROS.