玉米小斑病菌毒素诱导自交系Mo17凋亡的检测

Detection of apoptosis in Mo17 induced by Helminthosporium maydis race C(HMC) toxin

利用提取的玉米小斑病菌C小种(Helminthosporium maydis race C,HMC)毒素处理同核异质体Mo17-C和Mo17-N叶片及叶片中的原生质体并进行凋亡检测。叶片凋亡检测采用DNA凝胶电泳分析,结果表明:Mo17-C和Mo17-N均出现DNA-ladder,其中HMC毒素质量浓度为250μg/mL时,诱导12 h后Mo17-C出现的DNA-ladder最明显,而Mo17-N在HMC毒素质量浓度升至300μg/mL时,诱导12 h后DNA-ladder才最明显。原生质体的凋亡检测采用Hoechst 33258荧光染色法并统计其凋亡率,结果表明:Mo17-C和Mo17-N均出现细胞凋亡的特征,包括产生凋亡小体、环状染色质和染色质边集的现象,且凋亡率均随毒素浓度的增加和诱导时间的延长而呈上升趋势。但对于相同处理,C细胞质的凋亡率显著高于N细胞质,Mo17-C对HMC毒素的敏感性大于Mo17-N。

Apoptosis in leaves and protoplasts from homokaryon maize Mo17-C and Mo17-N treated by Helminthosporium maydis race C（HMC） toxin were detected by DNA ladder and hoechest 33258 respectively.The DNA ladder result indicates that DNA ladder appeared in both Mo17-C and Mo17-N,and the obvious DNA ladder in Mo17-C appeared after 12 h induction when the concentration of HMC reached 250 μg/mL,but appeared in Mo17-N when the concentration reached 300 μg/mL.Protoplasts of both Mo17-C and Mo17-N revealed apoptotic features such as apoptotic bodies,ring chromatin or chromatin margination.The apoptosis rate of Mo17-C maize protoplasts were significantly higher than that of Mo17-N in the same treatment,and it increased with the increase of treatment time and concentration.So,it is concluded that Mo17-C was more sensitive than Mo17-N.