Annexin V法检测盐胁迫下烟草原生质体的早期凋亡

摘要

将膜联蛋白V( Annexin V)法与碘化丙啶(PI)染色法相结合检测烟草(Nicotiana tabacum L.)原生质体的早期凋亡,结果表明:在中性盐胁迫下,能够很好地区分早期凋亡的原生质体和死亡的原生质体.200 mmol·L-1 NaC1对烟草原生质体胁迫5h,凋亡率能达到7.57％；但在200 mmol·L-1 NaHCO3碱性盐胁迫下,质膜透性发生了改变,不易区分早期凋亡的原生质体和死亡的原生质体,同时,随着碱性的增加,原生质体破碎率显著升高.碱性的NaHCO3和Na2CO3表现出对细胞膜的破坏作用.%An experiment was conducted to test the early apoptosis of tobacco ( Nicotiana tabacum L. ) protoplast using Annexin V-fluorescein isothiocyanate/propidium iodide staining. Results showed that, under the stress of neutral salt, early apoptosis and death of protoplasts could be well distinguished. And the apoptosis rate was 7. 57% under 200 mmol·L-1 NaCl stress for 5 hours. However, under the stress of 200 mraol·L-1 alkaline salt (NaHCO3) , the membrane permeability increased , and the early apoptosis and death of protoplasts couldn' t be well distinguished. The broken rate of protoplasts significantly increased with alkaline, which indicated the destructive effect of alkaline salt on membrane.