生物技术通报 ›› 2026, Vol. 42 ›› Issue (1): 86-94.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0542

• 研究报告 • 上一篇    下一篇

甲基汞胁迫下水稻OsMATE34的表达及功能分析

杨跃琴1(), 邢英1(), 仲子荷2,3, 田维军3,4, 杨雪清3,4, 王建旭3   

  1. 1.贵州师范大学化学与材料科学学院,贵阳 550025
    2.爱丁堡大学地球科学学院,爱丁堡 EH89YL 英国
    3.中国科学院地球化学研究所环境地球化学国家重点实验室,贵阳 550081
    4.中国科学院大学,北京 100049
  • 收稿日期:2025-05-26 出版日期:2026-01-26 发布日期:2026-02-04
  • 通讯作者: 邢英,女,博士,教授,研究方向 :重金属土壤环境化学;E-mail: xy31034@163.com
  • 作者简介:杨跃琴,女,硕士研究生,研究方向 :环境污染化学;E-mail: yangyueqin3569@163.com
  • 基金资助:
    国家自然科学基金项目(42222305);国家自然科学基金项目(42162008);贵州省重大科技专项项目(黔科合重大[2025]002);贵州省“百千万人才引进计划”第八批“千人创新创业人才”项目(GZQ202208090);贵州师范大学学术新苗项目黔师新苗[2022]25号

Expression and Functional Analysis of OsMATE34 in Rice under Mercury Stress

YANG Yue-qin1(), XING Ying1(), ZHONG Zi-he2,3, TIAN Wei-jun3,4, YANG Xue-qing3,4, WANG Jian-xu3   

  1. 1.School of Chemistry and Materials Science, Guizhou Normal University, Guiyang 550025
    2.School of Earth Sciences, University of Edinburgh, Edinburgh EH89YL, UK
    3.State Key Laboratory of Environmental Geochemistry, Institute of Geochemistry, Chinese Academy of Sciences, Guiyang 550081
    4.University of Chinese Academy of Sciences, Beijing 100049
  • Received:2025-05-26 Published:2026-01-26 Online:2026-02-04

摘要:

目的 多药和有毒化合物外排转运蛋白(MATE)在植物营养物质吸收、次生代谢物转运、重金属和外源物质解毒等方面发挥重要的调控作用。研究水稻OsMATE34的生物学功能,为深入理解水稻富集MeHg的分子机制奠定基础,亦为培育低MeHg水稻品种提供科学参考。 方法 通过生物信息学方法,分析OsMATE34的理化性质和系统进化树,利用实时荧光定量PCR(RT-qPCR)检测MeHg胁迫下水稻不同组织OsMATE34的相对表达量,并研究MeHg胁迫后水稻不同组织MeHg含量,用以分析其表达量与MeHg含量之间的相关性。利用CRISPR/Cas9基因编辑技术,获得1个OsMATE34敲除株系,在苗期进行MeHg处理,测定野生型(WT)和osmate34水稻中MeHg含量,探究OsMATE34对水稻MeHg积累的影响。 结果 水稻OsMATE34基因位于8号染色体上,编码489个氨基酸,其编码蛋白的相对分子量为51.60 kD,理论等电点(isoelectric point, pI)为5.6,为疏水性蛋白。RT-qPCR结果显示,MeHg胁迫抑制OsMATE34在根部表达;叶鞘和叶中OsMATE34表达量随着MeHg浓度升高显著上调,其中80 nmol/L甲基汞处理时叶鞘中基因表达较对照上调20倍。随着MeHg浓度升高,水稻不同组织MeHg含量均呈上升趋势,根中MeHg含量远高于叶鞘和叶。水稻叶鞘和叶中OsMATE34基因的表达量与MeHg含量随着MeHg胁迫浓度升高呈显著的正相关关系。与野生型相比,OsMATE34敲除株系叶鞘、叶和木质部MeHg含量均显著下降,根中MeHg含量无显著差异。 结论 OsMATE34基因参与MeHg由根向地上部的转运,为后续研究水稻吸收转运MeHg的分子机制奠定基础。

关键词: 甲基汞, 水稻, OsMATE34, 表达分析

Abstract:

Objective The multidrug and toxic compound extrusion transporter (MATE) plays a significant regulatory role in plant nutrient absorption, secondary metabolite transport, and detoxification of heavy metals and exogenous substances. Studying biological function of OsMATE34 in rice (Oryza sativa) may lay a foundation for a deeper understanding of the molecular mechanisms underlying MeHg accumulation in rice, and provide scientific reference for the breeding of low-MeHg rice varieties. Method Bioinformatics tools were used to analyze the physicochemical properties and phylogenetic tree of OsMATE34. Real-time quantitative PCR (RT-qPCR) was used to detect the relative expressions of OsMATE34 in different rice tissues under MeHg stress. The MeHg contents in different tissues of rice after MeHg exposure were measured, and the correlation between gene expressions and MeHg content was analyzed. A CRISPR/Cas9-mediated OsMATE34 knockout line was generated, and MeHg treatment was performed on seedlings. The MeHg contents in wild-type (WT) and osmate34 rice lines were measured to explore the effects of OsMATE34 on MeHg absorption and transport. Result The OsMATE34 gene was located on chromosome 8, encoding a protein of 489 amino acids with a relative molecular weight of 51.60 kD and a theoretical isoelectric point (pI) of 5.6. The protein was hydrophobic. RT-qPCR showed that MeHg stress inhibited the expression of OsMATE34 in roots, The expressions of OsMATE34 in the leaf sheath and leaves were significantly up-regulated with the increase of MeHg concentration, and the gene expression in stems was 20-fold higher than that of the control when treated with 80 nmol/L MeHg. The MeHg contents in rice tissues increased under MeHg stress, with the root accumulating significantly higher MeHg than the leaf sheath and leaf. There was a significant positive correlation between OsMATE34 gene expression and MeHg content in rice leaf sheath and leaves with the increase of MeHg concentration. Compared to the wild-type, the OsMATE34 knockout lines showed a significant reduction in MeHg content in the leaf sheath, leaf, and xylem, while there was no significant difference in the root. Conclusion The OsMATE34 gene is involved in the transport of MeHg from the root to the aboveground tissues. This study provides a foundation for further research on the molecular mechanisms of MeHg absorption and transport in rice.

Key words: MeHg, rice, OsMATE34, expression analysis