Department of Plant, Soil and Insect Sciences, University of Massachusetts, 202 French Hall, Amherst, MA 01003, USA.
Transgenic Res. 2012 Dec;21(6):1265-77. doi: 10.1007/s11248-012-9600-8. Epub 2012 Feb 19.
Rice accumulates high level of arsenic (As) in its edible parts and thus plays an important role in the transfer of As into the food chain. However, the mechanisms of As uptake and its detoxification in rice are not well understood. Recently, members of the Nodulin 26-like intrinsic protein (NIP) subfamily of plant aquaporins were shown to transport arsenite in rice and Arabidopsis. Here we report that members of the rice plasma membrane intrinsic protein (PIP) subfamily are also involved in As tolerance and transport. Based on the homology search with the mammalian AQP9 and yeast Fps1 arsenite transporters, we identified and cloned five rice PIP gene subfamily members. qRT-PCR analysis of PIPs in rice root and shoot tissues revealed a significant down regulation of transcripts encoding OsPIP1;2, OsPIP1;3, OsPIP2;4, OsPIP2;6, and OsPIP2;7 in response to arsenite treatment. Heterologous expression of OsPIP2;4, OsPIP2;6, and OsPIP2;7 in Xenopus laevis oocytes significantly increased the uptake of arsenite. Overexpression of OsPIP2;4, OsPIP2;6, and OsPIP2;7 in Arabidopsis yielded enhanced arsenite tolerance and higher biomass accumulation. Further, these transgenic plants showed no significant accumulation of As in shoot and root tissues in long term uptake assays. Whereas, short duration exposure to arsenite caused both active influx and efflux of As in the roots. The data suggests a bidirectional arsenite permeability of rice PIPs in plants. These rice PIPs genes will be highly useful for engineering important food and biofuel crops for enhanced crop productivity on contaminated soils without increasing the accumulation of toxic As in the biomass or edible tissues.
水稻在可食用部分积累了高水平的砷(As),因此在将 As 转移到食物链中起着重要作用。然而,水稻吸收 As 的机制及其解毒机制还不是很清楚。最近,植物水通道蛋白的 Nodulin 26 样内在蛋白(NIP)亚家族的成员被证明可以在水稻和拟南芥中转运亚砷酸盐。在这里,我们报告水稻质膜内在蛋白(PIP)亚家族的成员也参与了 As 耐受和转运。根据与哺乳动物 AQP9 和酵母 Fps1 亚砷酸盐转运蛋白的同源性搜索,我们鉴定并克隆了五个水稻 PIP 基因亚家族成员。对水稻根和茎叶组织中 PIPs 的 qRT-PCR 分析显示,OsPIP1;2、OsPIP1;3、OsPIP2;4、OsPIP2;6 和 OsPIP2;7 的转录本在受到亚砷酸盐处理时显著下调。OsPIP2;4、OsPIP2;6 和 OsPIP2;7 在非洲爪蟾卵母细胞中的异源表达显著增加了亚砷酸盐的摄取。在拟南芥中过表达 OsPIP2;4、OsPIP2;6 和 OsPIP2;7 提高了亚砷酸盐的耐受性和生物量的积累。此外,这些转基因植物在长期摄取试验中,茎叶组织中没有明显的 As 积累。然而,在亚砷酸盐的短期暴露下,根部既发生了 As 的主动内流又发生了外排。数据表明,水稻 PIPs 在植物中具有双向的亚砷酸盐通透性。这些水稻 PIP 基因对于在污染土壤上提高作物生产力而不增加生物量或可食用组织中有毒 As 的积累,对于工程重要的粮食和生物燃料作物非常有用。