INT J MOL SCI :西南大学丨杨树ABC转运蛋白的异位表达赋予拟南芥Cd耐受性(附NMT实验体系)
- 期刊:international journal of molecular sciences
- 主题:杨树ABC转运蛋白的异位表达赋予拟南芥Cd耐受性
- 标题:Ectopic Expression of Poplar ABC Transporter PtoABCG36 Confers Cd Tolerance in Arabidopsis thaliana
- 影响因子:4.183
- 检测指标: Cd2+流速
- 检测部位:拟南芥距离根尖800μm
- Cd2+流实验处理方法:15日龄幼苗,50μM CdCl2处理24小时
- Cd2+流实验测试液成份:0.1 mM KCl, 0.1 mM CaCl2, 0.05 mM CdCl2, 0.3 mM 2-(N-morpholino)ethane sulfonic acid, pH 5.8
- 通讯作者:西南大学王邦俊
英文摘要
Cadmium (Cd) is one of the most toxic heavy metals for plant growth in soil. ATP-binding cassette (ABC) transporters play important roles in biotic and abiotic stresses. However, few ABC transporters have been characterized in poplar.
In this study, we isolated an ABC transporter gene PtoABCG36 from Populus tomentosa. The PtoABCG36 transcript can be detected in leaves, stems and roots, and the expression in the root was 3.8 and 2 times that in stems and leaves, respectively. The PtoABCG36 expression was induced and peaked at 12 h after exposure to Cd stress. Transient expression of PtoABCG36 in tobacco showed that PtoABCG36 is localized at the plasma membrane. When overexpressed in yeast and Arabidopsis, PtoABCG36 could decrease Cd accumulation and confer higher Cd tolerance in transgenic lines than in wild-type (WT) lines. Net Cd2+ efflux measurements showed a decreasing Cd uptake in transgenic Arabidopsis roots than WT.
These results demonstrated that PtoABCG36 functions as a cadmium extrusion pump participating in enhancing tolerance to Cd through decreasing Cd content in plants, which provides a promising way for making heavy metal tolerant poplar by manipulating ABC transporters in cadmium polluted areas.
中文摘要(谷歌机翻)
镉(Cd)是土壤中植物生长最毒的重金属之一。 ATP结合盒(ABC)转运蛋白在生物和非生物胁迫中起重要作用。然而,很少有ABC转运蛋白在杨树中具有特征。
在这项研究中,我们从毛白杨(Populus tomentosa)中分离出一个ABC转运蛋白基因PtoABCG36。可以在叶,茎和根中检测到PtoABCG36转录物,并且根中的表达分别是茎和叶中的3.8和2倍。诱导PtoABCG36表达并在暴露于Cd胁迫后12小时达到峰值。 PtoABCG36在烟草中的瞬时表达显示PtoABCG36位于质膜上。当在酵母和拟南芥中过表达时,PtoABCG36可以减少Cd积累并赋予转基因品系中比野生型(WT)品系更高的Cd耐受性。净Cd2+外排测量显示转基因拟南芥根中的Cd摄取减少比WT。
这些结果表明,PtoABCG36作为镉挤压泵发挥作用,通过降低植物中的Cd含量来增强对Cd的耐受性,这为通过操纵镉污染区域的ABC转运蛋白制备重金属耐受性杨树提供了一种有前景的方法。
结果表明:在50 M CdCl2存在下,OX-1,OX-2和OX-3系的Cd2+净吸收量低于WT植物(分别为62.39%,54.50%和53.30%)。 相比之下,突变体abcg36具有比WT植物更高的Cd2+净吸收量。
文章链接:https://www.mdpi.com/1422-0067/20/13/3293