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【目的】研究玉米小斑病菌C小种毒素(HMC-toxin)诱导同核异质体玉米的离体根冠细胞发生凋亡的差异性及凋亡规律。【方法】采用吖啶橙(AO)与溴化乙啶(EB)荧光复染以及Hoechst 33258荧光染色的方法检测。【结果】HMC毒素诱导根冠细胞发生凋亡,出现凋亡小体与染色体边集形态特征。在3种毒素浓度、3种处理时间下,Mo17-C的根冠细胞死亡率均高于Mo17-N。采用AO与EB复染方法,在150μg.mL-1HMC毒素处理7 h时,Mo17-C细胞凋亡率达最高值,为68.7%,而Mo17-N仅为29%;经Hoechst 33258染色后,在150μg.mL-1HMC毒素处理下,Mo17-C也于7 h时达最大凋亡率,为57.3%,而Mo17-N为27.7%。【结论】在毒素“伤害”细胞致死过程中,C细胞质的细胞死亡率远高于N细胞质;在“诱导”细胞凋亡程序中,C细胞质的细胞凋亡率也远高于N细胞质的。凋亡率随诱导浓度和诱导时间的增加而增加。AO、EB复染与Hoechst 33258染色相比,两者细胞凋亡率接近,准确度均高,图像清晰,且费用相当,但前者可以区分早期和晚期凋亡细胞而后者不能区分。
【Objective】 The purpose of this study was to investigate the difference in apoptosis and apoptosis of isolated root-and-crown cells induced by HMC-toxin in corn co-nuclear heterotrophic maize. 【Method】 Fluorescent staining with acridine orange (AO) and ethidium bromide (EB) and fluorescence staining with Hoechst 33258 were used. 【Result】 HMC toxin induced apoptosis in root and crown cells, and apoptotic body and chromosome border features were observed. At three toxin concentrations and three treatment times, the root-cell death rate of Mo17-C was higher than that of Mo17-N. The apoptotic rate of Mo17-C cells reached the highest value at68.7% and Mo17-N was only29% when treated with 150μg.mL-1HMC toxin for7h by AO and EB staining. After Hoechst 33258 staining, Mo17-C also reached the maximum apoptotic rate of 57.3% at 7 h and the Mo17-N was 27.7% under the treatment of 150 μg.mL-1HMC toxin. 【Conclusion】 The cell death rate of C cytoplasm is much higher than that of N cytoplasm during the lethality of toxin “injury ”. The rate of apoptosis of C cytoplasm is much higher than that of N N cytoplasmic. Apoptosis rate increased with the induction concentration and induction time. Compared with Hoechst 33258 staining, the apoptotic rates of both AO and EB were close and the accuracy was high, the images were clear and the cost was the same, but the former could distinguish the early and late apoptotic cells and the latter could not be distinguished.