流式细胞仪检测铝胁迫诱导的花生悬浮细胞程序性死亡  

Aluminum induced programed cell death of peanut suspension cultures detected by flow cytometry

作　　者：黄文静[1] 何虎翼[1] 邓伦武[1] 王爱勤[1] 李创珍[1] 韦善清[1] 何龙飞[1] 

机构地区：[1]广西大学农学院,广西南宁530004

出　　处：《中国油料作物学报》2014年第1期51-58,共8页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金资助项目(31260296;30960181)

摘　　要：以耐铝品种99-1507和铝敏感品种中花2号为材料,应用荧光显微镜和流式细胞仪研究铝对悬浮细胞活性及其线粒体生理的影响,并检测悬浮细胞程序性死亡率。结果表明:铝处理12h后,随着铝浓度增加,悬浮细胞FDA(Fluorescein diacetate)荧光信号逐渐减弱,PI(Propidium Iodide)荧光信号增强,细胞活性下降;线粒体超氧阴离子含量和H2O2含量随着铝浓度的增加逐渐上升;线粒体膜电位(Δψm)随着铝浓度增加逐渐下降,中花2号下降幅度大于99-1507。50μmol/L及其以上浓度的铝可以显著诱导花生悬浮细胞程序性死亡,铝浓度越高死亡细胞数量越多,同等浓度铝处理下中花2号细胞死亡率高于99-1507,差异达到显著水平。应用流式细胞仪能够准确地检测悬浮细胞程序性死亡的发生情况,细胞程序性死亡率与花生品种的耐铝性呈负相关关系。The suspension cell apoptotic rate and mitochondria viability of two peanut varieties （99 -1507 Al - tolerant and ZH2 A1 -sensitive） treated by different concentration aluminum （0,20,100,400 μmol/L） were examined by fluorescence microscope and flow cytometry. The results showed that after A1 treatment for 12h, the fluorescent light produced by FDA （Fluorescein diacetate）decreased, but increased by PI （Propidium Iodide）with the increase of A1 concentration, which indicated that peanut cytoactive decreased. Content of O2 ＇ - and H2O2 in mitochondria increased, and both mitochondrial membrane potential of two peanut varieties decreased, decrement of ZH2 was higher than 99 - 1507. 50μmol/L and above A1 could significantly induce peanut suspension cultures pro- gramed cell death. With the increase of A1 concentration, the number of programed death cell induced by A1 increased. The number was higher in ZH2 than in 99 - 1507. There were significant differences between the two peanut varieties. By using flow cytometry, the apoptotic of peanut suspension cell could be inspected accurately. There was a negative correlation between cell apoptpsis rate and A1 - resistance of peanut.

关 键 词：花生 悬浮细胞 铝胁迫 细胞程序性死亡 流式细胞仪 线粒体膜电位 

分 类 号：S565.201[农业科学—作物学] S311