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激发子 PB90 是疫霉菌(Phytophthora boehmeriae)分泌的一种分子量为90 kD 的胞外蛋白激发子,该激发子可以诱导非寄主植物烟草的过敏反应和系统获得抗性. BY-2烟草悬浮细胞经该激发子PB90处理后, Trypan blue 染色观察到细胞死亡, 这种死亡可被蛋白质合成抑制剂 Cycloheximide 抑制, 表明这种细胞死亡是主动死亡过程. 提取 PB90 处理的悬浮细胞的 DNA 进行琼脂糖凝胶电泳观察到 DNALadder 现象, 表明核小体间的 DNA 发生了断裂; 对经 PB90 分别处理 4 和 12 h 的烟草悬浮细胞进行TUNEL 染色, 观察到较强的阳性信号, 进一步表明其细胞核内发生了 DNA 末端断裂; 同时结合 DAPI染色观察到经 PB90 处理后的细胞染色质凝集、细胞核解体形成凋亡小体等细胞凋亡的形态学特征. 上述结果表明激发子 PB90 可以诱导 BY-2 烟草悬浮细胞发生细胞凋亡, 提示该激发子诱导的过敏反应是一个细胞凋亡过程.
The elicitor PB90 is an extracellular protein exciton secreted by Phytophthora boehmeriae with a molecular weight of 90 kD, which can induce allergic reactions and systemic acquired resistance in non-host plant tobacco BY-2 Tobacco suspension cells Cell death was observed by Trypan blue staining after PB90 treatment, and this death was inhibited by the protein synthesis inhibitor Cycloheximide, indicating that this cell death was the active death process. The DNA of PB90-treated suspension cells was extracted and subjected to agarose gel The DNALadder phenomenon was observed by electrophoresis, which indicated that the nucleosome DNA was cleaved. TUNEL staining of tobacco suspension cells treated with PB90 for 4 and 12 h, respectively, showed a strong positive signal, which further indicated that DNA occurred in the nucleus End breakage.At the same time, the morphological characteristics of apoptotic bodies such as chromatin agglutination and cell nucleus disintegration were observed by DAPI staining.The results indicated that PB90 could induce the production of BY-2 tobacco suspension cells Apoptosis, suggesting that the elicitor-induced allergic reaction is a apoptosis Cheng.