摘要
该实验以烟草悬浮细胞BY-2为材料,在烟草悬浮细胞中分别加入0.05、0.10、0.15、0.20mmol·L^-1AlCl3,以等体积去离子水处理的悬浮细胞液为对照,并依据前述实验结果选择0.15mmol·L^-1AlCl3,分别添加5mmol·L^-1DMTU(H2O2抑制剂)、20μmol·L^-1CaCl2、15μmol·L^-1LaCl3(Ca^2+通道抑制剂)和50μmol·L^-1ATP设计多项处理,分析胞外ATP(eATP)对铝离子(Al^3+)胁迫引起的植物细胞死亡及其胞内H2O2、Ca^2+的影响,以揭示Al3+胁迫下植物调节细胞死亡的可能机制,进一步扩展对eATP功能的认知。结果显示:(1)随着AlCl3胁迫浓度的提高,细胞死亡水平和胞内H2O2水平上升,而胞内Ca^2+和eATP水平则逐渐降低。(2)外援施加H2O2抑制剂DMTU(二甲基硫脲)和Ca^2+能够有效缓解AlCl3诱导的细胞死亡水平的上升;而Ca^2+通道抑制剂LaCl3(三氯化镧)则加剧了AlCl3胁迫下的细胞死亡。(3)在AlCl3胁迫下对细胞添加外源ATP,能够缓解AlCl3胁迫下胞内H2O2水平上升和Ca^2+水平下降的同时,并显著降低AlCl3胁迫导致的细胞死亡。研究表明,Al^3+以剂量依赖的模式提升细胞死亡和细胞内H2O2的水平并降低胞内Ca^2+和eATP水平,AlCl3诱导的细胞死亡受到H2O2和Ca^2+水平变化的调节,eATP可以通过调节H2O2与Ca^2+水平缓解AlCl3诱导的细胞死亡。推测Al3+胁迫可能通过抑制钙离子通道而破坏了细胞内H2O2和Ca^2+之间的协同关系,外源ATP对Al^3+诱导H2O2上升的缓解作用可能是由于其提升了细胞的抗氧化能力。
In this experiment,tobacco suspension cell BY-2 was used as the material,and 0.05,0.10,0.15,0.20 mmol·L^-1 AlCl3 was added to the tobacco suspension cells,and the suspension cell solution treatment with equal volume of deionization water was used as a control.And,according to the above experimental results,0.15 mmol·L^-1 AlCl3 was selected,5 mmol·L^-1 DMTU(H2O2 inhibitor),20μmol·L^-1 CaCl2,15μmol·L^-1 LaCl3(Ca^2+channel inhibitor)and 50μmol·L-1 ATP was used to analyze the effects of extra-cellular ATP(eATP)on plant cell death and intra-cellular H2O2 and Ca^2+induced by aluminum ion(Al^3+)stress and to reveal plant the possible mechanism of eATP in regulating cell death under Al^3+stress,and further extend the perception of eATP function.The results showed that:(1)with the increase of AlCl3 concentration,the levels of cell death and intracellular H 2O 2 were increased,while the levels of intracellular Ca^2+and eATP were decreased;(2)Application of DMTU(dimethylthiourea,a scavenger of H 2O 2)and exogenous Ca^2+effectively attenuated the AlCl3-induced cell death,while barium trichloride(LaCl3,the Ca^2+channel inhibitor)aggravated the AlCl3-induced cell death,indicating that AlCl3-induced cell death is regulated by H2O2 and Ca^2+;(3)The addition of exogenous ATP effectively impeded the increase of intracellular H2O2 production and decrease of intracellular Ca^2+under AlCl^3 stress,and the cell death induced by AlCl 3 were also alleviated by exogenous ATP.It can be seen that aluminum-induced cell death is regulated by changes in H2O2 and Ca^2+levels,and extracellular ATP affects the aluminum-induced cell death by regulating H2O2 and Ca^2+levels.Theses observations indicated that the level of cell death and intracellular H2O2 were increased,while the level of intracellular Ca^2+and eATP were decreased in a dose-dependent manner under Al^3+stress.It can be seen that aluminum-induced cell death is regulated by changes in H2O2 and Ca^2+levels,and extracellular ATP affects the aluminum-induced cell death by regula
作者
达梦婷
石珍珍
庞海龙
贾凌云
孙坤
冯汉青
DA Mengting;SHI Zhenzhen;PANG Hailong;JIA Lingyun;SUN Kun;FENG Hanqing(College of Life Science,Northwest Normal University,Lanzhou,730070,China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2019年第6期1033-1041,共9页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(31870246,31560070)
甘肃省高等学校科研项目(2015A-007)
甘肃省重点研发计划(18YF1NA051)
甘肃省高校基本科研业务费
西北师范大学青年创新团队项目
