Department of Public Health, School of Pharmacy, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Plant Cell Physiol. 2017 Oct 1;58(10):1730-1742. doi: 10.1093/pcp/pcx114.
Phytochelatin (PC) synthesis has been well demonstrated as a major metal tolerance mechanism in Arabidopsis thaliana, whereas its contribution to long-distance element transport especially in monocots remains elusive. Using rice as a cereal model, we examined physiological roles of Oryza sativa phytochelatin synthase 1 (OsPCS1) in the distribution and detoxification of arsenic (As) and cadmium (Cd), two toxic elements associated with major food safety concerns. First, we isolated four different transcript variants of OsPCS1 as well as one from OsPCS2. Quantitative real-time reverse transcription-PCR (RT-PCR) of each OsPCS transcript in rice seedlings suggested that expression of OsPCS1full, the longest OsPCS1 variant, was most abundant, followed by OsPCS2. Heterologous expression of OsPCS variants in PCS-deficient mutants of Schizosaccharomyces pombe and A. thaliana suggested that OsPCS1full possessed PCS activity in response to As(III) and Cd while the activity of other PCS variants was very low. To address physiological functions in toxic element tolerance and accumulation, two independent OsPCS1 mutant rice lines (a T-DNA and a Tos17 insertion line) were identified. The OsPCS1 mutants exhibited increased sensitivity to As(III) and Cd in hydroponic experiments, showing the importance of OsPCS1-dependent PC synthesis for rice As(III) and Cd tolerance. Elemental analyses of rice plants grown in soil with environmentally relevant As and Cd concentrations showed increased As accumulation and decreased Cd accumulation in grains of the T-DNA line. The Tos17 mutant also exhibited the reduced Cd accumulation phenotype. These contrasting effects on As and Cd distribution to grains suggest the existence of at least partially distinct PC-dependent pathways for As and Cd.
植物螯合肽(PC)的合成已被证明是拟南芥金属耐受的主要机制,然而,其在单子叶植物中的长距离元素运输中的贡献仍然难以捉摸。本研究以水稻为谷类模式,研究了水稻谷胱甘肽合成酶 1(OsPCS1)在砷(As)和镉(Cd)分布和解毒中的生理作用,As 和 Cd 是与主要食品安全问题相关的两种有毒元素。首先,我们从 OsPCS2 中分离出四种不同的 OsPCS1 转录变体和一种。水稻幼苗中每个 OsPCS 转录物的定量实时 RT-PCR(qRT-PCR)表明,OsPCS1full 的表达最为丰富,OsPCS1full 是最长的 OsPCS1 变体,其次是 OsPCS2。在酿酒酵母和拟南芥 PCS 缺陷突变体中异源表达 OsPCS 变体表明,OsPCS1full 在响应 As(III)和 Cd 时具有 PCS 活性,而其他 PCS 变体的活性非常低。为了研究在有毒元素耐受和积累中的生理功能,鉴定了两个独立的 OsPCS1 突变体水稻系(一个 T-DNA 和一个 Tos17 插入系)。在水培实验中,OsPCS1 突变体对 As(III)和 Cd 的敏感性增加,表明 OsPCS1 依赖的 PC 合成对水稻 As(III)和 Cd 耐受的重要性。在土壤中生长的水稻植株的元素分析表明,在含有环境相关 As 和 Cd 浓度的土壤中,T-DNA 系的 As 积累增加,而 Cd 积累减少。Tos17 突变体也表现出减少的 Cd 积累表型。这些对 As 和 Cd 向谷物分布的相反影响表明,至少存在部分不同的 PC 依赖途径用于 As 和 Cd。