Search for genes that confer acid tolerance to plants

T. Aoki, M. Shimamura, Y. Sawada, R. Imaizumi
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Abstract

To remediate the worldwide-spreading acid sulphate soil area and utilize it for human beings, acid tolerant leguminous plants are expected to be useful as crops and pioneer plants. The purpose of this study is to produce symbiotic nitrogen fixing systems tolerant to acid sulfate soils, through the identification of the genes that confer acid tolerance on plants and the isolation of acid-tolerant rhizobia. We identified acid tolerant genes by two approaches. One approach is functional screening of a soybean cDNA library and their characterization in planta. E. coli cells were transformed with a cDNA library of soybean and cultured on a LB medium adjusted to pH 4 or pH 5. Selected six tolerant clones were introduced into Arabidopsis thaliana using a plant expression vector. The overexpressors grew better than the wild type under the acid and aluminium stresses. The other one is molecular genetic analysis of the candidate genes identified by previous studies: a gene for a key enzyme of sulfur assimilation, cysteine synthase. The candidate gene was ectopically overexpressed in A. thaliana under the control of the cauliflower mosaic virus 35S promoter. All the genes tested were shown to confer acid tolerance on the model plants.
寻找赋予植物耐酸能力的基因
为了修复世界范围内广泛分布的酸性硫酸盐土壤,使其为人类所利用,耐酸豆科植物有望成为有益的作物和先锋植物。本研究的目的是通过鉴定赋予植物耐酸基因和分离耐酸根瘤菌,产生耐酸性硫酸盐土壤的共生固氮系统。我们通过两种方法鉴定耐酸基因。一种方法是大豆cDNA文库的功能筛选及其在植物中的鉴定。用大豆cDNA文库转化大肠杆菌细胞,并在调节pH为4或5的LB培养基上培养。选择6个耐受性无性系,利用植物表达载体导入拟南芥。在酸和铝胁迫下,过表达型比野生型生长得更好。二是对硫同化关键酶半胱氨酸合成酶基因进行分子遗传分析。该候选基因在花椰菜花叶病毒35S启动子控制下在拟南芥中异位过表达。所有测试的基因都显示出模式植物具有耐酸性。
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