College of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming, Yunnan, China.
State Key Laboratory of Conservation and Utilization for Bioresources in Yunnan, Yunnan University, Kunming, Yunnan, China.
BMC Plant Biol. 2020 Mar 20;20(1):124. doi: 10.1186/s12870-020-2316-7.
Panax notoginseng is a medicinally important Chinese herb with a long history of cultivation and clinical application. The planting area is mainly distributed in Wenshan Prefecture, where the quality and safety of P. notoginseng have been threatened by high concentration of arsenic (As) from the soil. The roles of phosphate (Pi) transporters involved in Pi acquisition and arsenate (AsV) tolerance were still unclear in this species.
In this study, two open reading frames (ORFs) of PnPht1;1 and PnPht1;2 separated from P. notoginseng were cloned based on RNA-seq, which encoded 527 and 541 amino acids, respectively. The results of relative expression levels showed that both genes responded to the Pi deficiency or As exposure, and were highly upregulated. Heterologous expression in Saccharomyces cerevisiae MB192 revealed that PnPht1;1 and PnPht1;2 performed optimally in complementing the yeast Pi-transport defect, particularly in PnPht1;2. Cells expressing PnPht1;2 had a stronger AsV tolerance than PnPht1;1-expressing cells, and accumulated less As in cells under a high-Pi concentration. Combining with the result of plasma membrane localization, these data confirmed that transporters PnPht1;1 and PnPht1;2 were putative high-affinity H/HPO symporters, mediating the uptake of Pi and AsV.
PnPht1;1 and PnPht1;2 encoded functional plasma membrane-localized transporter proteins that mediated a putative high-affinity Pi/H symport activity. Expression of PnPht1;1 or PnPht1;2 in mutant strains could enhance the uptake of Pi and AsV, that is probably responsible for the As accumulation in the roots of P. notoginseng.
三七是一种药用历史悠久的重要中草药,其种植区主要分布在文山州,这里的土壤砷浓度较高,威胁着三七的质量和安全。目前,参与磷获取和砷酸盐(AsV)耐受的磷酸(Pi)转运体的作用在该物种中仍不清楚。
本研究根据 RNA-seq 从三七中克隆了两个开放阅读框(ORFs)PnPht1;1 和 PnPht1;2,分别编码 527 和 541 个氨基酸。相对表达水平的结果表明,这两个基因都对缺磷或砷暴露有反应,并高度上调。在酿酒酵母 MB192 中的异源表达表明,PnPht1;1 和 PnPht1;2 最能互补酵母 Pi 转运缺陷,尤其是在 PnPht1;2 中。表达 PnPht1;2 的细胞比表达 PnPht1;1 的细胞具有更强的 AsV 耐受性,并且在高 Pi 浓度下细胞中积累的 As 较少。结合质膜定位的结果,这些数据证实了转运体 PnPht1;1 和 PnPht1;2 是假定的高亲和力 H/HPO 共转运体,介导 Pi 和 AsV 的摄取。
PnPht1;1 和 PnPht1;2 编码功能性质膜定位转运蛋白,介导假定的高亲和力 Pi/H 共转运活性。在突变株中表达 PnPht1;1 或 PnPht1;2 可以增强 Pi 和 AsV 的摄取,这可能是三七根系中 As 积累的原因。