Expression of Arabidopsis acyl-CoA-binding proteins AtACBP1 and AtACBP4 confers Pb(II) accumulation in Brassica juncea roots
Expression of Arabidopsis acyl-CoA-binding proteins AtACBP1 and AtACBP4 confers Pb(II) accumulation in Brassica juncea roots
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DOI：
10.1111/pce.12382
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发表时间：
2015-01-01
影响因子：
7.3
通讯作者：
Chye, Mee-Len
中科院分区：
文献类型：
--
作者：
Du, Zhi-Yan;Chen, Mo-Xian;Chye, Mee-Len
在拟南芥中，在qRT-PCR分析中观察到两个编码酰基辅酶A结合蛋白（ACBPs）的基因AtACBP 1和AtACBP 4的表达被铅[Pb（II）]诱导。GUS（β-葡萄糖醛酸苷酶）活性的定量测定证实了Pb（II）对AtACBP 1 pro：：GUS的诱导作用。电泳迁移率变动分析（EMSAs）显示，在AtACBP 1的5-侧翼区的Pas元件响应Pb（II）处理。AtACBP 1和AtACBP 4在Pb（II）的吸收利用芥菜，一个潜在的候选植物修复由于其快速增长，大根，高生物量和良好的能力，积累重金属进一步比较。原子吸收光谱分析结果表明，转基因B.juncea表达AtACBP 1或AtACBP 4表明Pb（II）积累在根中。随后的铅（II）示踪试验表明，Pb（II）积累在根尖和维管组织的转基因B.junceaAtACBP1-overexpressors（OXs）和AtACBP 4-OXs和转基因acetopsisAtACBP 1-OXs的胞质溶胶。转基因拟南芥AtACBP 1-OXs螯合Pb（II）的毛状体，并显示耐受过氧化氢（H2 O2）处理。此外，AtACBP 1和AtACBP 4在野生型拟南芥的根中被H2 O2诱导，而B.junceaAtACBP1-OX和AtACBP 4-OX根的脂质过氧化氢（LOOH）测量表明AtACBP 1和AtACBP 4可以保护脂质免受Pb（II）诱导的脂质过氧化。拟南芥酰基辅酶A结合蛋白AtACBP 1和AtACBP 4已被证明在体外结合铅[Pb（II）]，促使我们研究它们在Pb（II）植物修复中的潜力。我们的研究结果表明，AtACBP 1和AtACBP 4的诱导铅（II）和推定的元件响应铅（II）在AtACBP 1的5-侧翼区被确定。过表达AtACBP 1的转基因拟南芥将Pb（II）隔离在毛状体中，并表现出对过氧化氢（H2 O 2）处理的耐受性，而过表达AtACBP 1或AtACBP 4的转基因芥菜将Pb（II）隔离在根尖和维管组织的胞质溶胶中，并表现出Pb（II）在根中的积累。此外，在野生型拟南芥和转基因B的根中，AtACBP 1和AtACBP 4被H2 O 2诱导表达。过表达AtACBP 1或AtACBP 4的芥菜对Pb（II）诱导的脂质过氧化有保护作用。
In Arabidopsis thaliana, the expression of two genes encoding acyl-CoA-binding proteins (ACBPs) AtACBP1 and AtACBP4, were observed to be induced by lead [Pb(II)] in shoots and roots in qRT-PCR analyses. Quantitative GUS (-glucuronidase) activity assays confirmed induction of AtACBP1pro::GUS by Pb(II). Electrophoretic mobility shift assays (EMSAs) revealed that Pas elements in the 5-flanking region of AtACBP1 were responsive to Pb(II) treatment. AtACBP1 and AtACBP4 were further compared in Pb(II) uptake using Brassica juncea, a potential candidate for phytoremediation given its rapid growth, large roots, high biomass and good capacity to accumulate heavy metals. Results from atomic absorption analyses on transgenic B.juncea expressing AtACBP1 or AtACBP4 indicated Pb(II) accumulation in roots. Subsequent Pb(II)-tracing assays demonstrated Pb(II) accumulation in the cytosol of root tips and vascular tissues of transgenic B.junceaAtACBP1-overexpressors (OXs) and AtACBP4-OXs and transgenic ArabidopsisAtACBP1-OXs. Transgenic ArabidopsisAtACBP1-OXs sequestered Pb(II) in the trichomes and displayed tolerance to hydrogen peroxide (H2O2) treatment. In addition, AtACBP1 and AtACBP4 were H2O2-induced in the roots of wild-type Arabidopsis, while lipid hydroperoxide (LOOH) measurements of B.junceaAtACBP1-OX and AtACBP4-OX roots suggested that AtACBP1 and AtACBP4 can protect lipids against Pb(II)-induced lipid peroxidation.Arabidopsis acyl-CoA-binding proteins, AtACBP1 and AtACBP4, have been shown to bind lead [Pb(II)] in vitro, prompting us to investigate their potential in Pb(II) phytoremediation. Our results showed that AtACBP1 and AtACBP4 were induced by Pb(II) and putative elements responsive to Pb(II) were identified in the 5-flanking region of AtACBP1. Transgenic Arabidopsis overexpressing AtACBP1 sequestered Pb(II) in the trichomes and displayed tolerance to hydrogen peroxide (H (2) O (2)) treatment, whereas transgenic Brassica juncea overexpressing AtACBP1 or AtACBP4 sequestered Pb(II) in the cytosol of root tips and vascular tissues, and displayed Pb(II) accumulation in roots. In addition, AtACBP1 and AtACBP4 were induced by H (2) O (2) in roots of wild-type Arabidopsis and transgenic B. juncea overexpressing AtACBP1 or AtACBP4 were protected against Pb(II)-induced lipid peroxidation.