Duan Jian-Gong, Li Hong-Yu, Ru Yi, Liang Hou-Guo, Wang Jin-Sheng, Li Min-Quan, Yang Fu-Jun
School of Life Sciences, Lanzhou University, Lanzhou 730000, China.
Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2005 Jun;31(3):322-6.
O(-.)(2) production based on chemical method by Ecc-treated intact tobacco leaves was observed. It showed a long-lasting one-phase course beginning 1 h and ending 14 h after Erwinia carotovora subsp. carotovora (Ecc) was inoculated. In Ecc-treated leaves, O(-.)(2) production rate of 3 h is 2 times of 1 h, and of 10 h it remains 1.7 times. It decreases rapidly between 10 h and 14 h, at 14 h it falls to the level before inoculation. This is a completely different course from the widely accepted two-phase course obtained from suspension-cultured cells. Electron spin resonance (ESR) spectrum of O(-.)(2) produced by intact tobacco leaves after Ecc inoculation 2 h and 6 h was also observed, the amplitude of 6 h is larger than of 2 h in Ecc-treated group. In the control group, there is no difference between the amplitude of 2 h and 6 h. This indicated that O(-.)(2) amount of Ecc-treated group is higher than of the control group, which is the same result as obtained through chemical method. The ESR spectrum of O(-.)(2) produced by chloroplasts and mitochondria from tobacco leaves after 2 h and 6 h after Ecc inoculation were observed. In chloroplast experiment, the amplitude of 6 h was larger than of 2 h in Ecc-treated group while it was the same in the control group. ESR spectrum of O(-.)(2) produced by mitochondria was proved to be a same result after a careful comparison, in spite of a greater difference between 2 h and 6 h in control group than chloroplast experiment. The fact that spectra of these two organelles were both synchronous with that of intact leaves implied that these two organelles possibly participated in cellular O(-.)(2) burst. Chloroplasts in the dark showed no ESR spectrum 2 h and 6 h after Ecc inoculation in Ecc-treated group as well as in the control group, indicating that O(-.)(2) in the chloroplast probably originated from the photosynthetic electron transport process.
观察了经胡萝卜软腐欧文氏菌胡萝卜软腐亚种(Ecc)处理的完整烟草叶片基于化学方法的超氧阴离子(O(-.)(2))产生情况。结果显示其呈现出一个持久的单相过程,在接种胡萝卜软腐欧文氏菌胡萝卜软腐亚种(Ecc)后1小时开始,14小时结束。在经Ecc处理的叶片中,3小时的O(-.)(2)产生速率是1小时的2倍,10小时时仍为1小时的1.7倍。在10小时至14小时之间其迅速下降,14小时时降至接种前水平。这与从悬浮培养细胞中获得的被广泛接受的两相过程完全不同。还观察了接种Ecc 2小时和6小时后完整烟草叶片产生的O(-.)(2)的电子自旋共振(ESR)光谱,在Ecc处理组中,6小时的光谱幅度大于2小时的。在对照组中,2小时和6小时的光谱幅度没有差异。这表明Ecc处理组的O(-.)(2)量高于对照组,这与通过化学方法获得的结果相同。观察了接种Ecc 2小时和6小时后烟草叶片叶绿体和线粒体产生的O(-.)(2)的ESR光谱。在叶绿体实验中,Ecc处理组6小时的光谱幅度大于2小时的,而对照组中两者相同。经过仔细比较,线粒体产生的O(-.)(2)的ESR光谱也得到了相同的结果,尽管对照组中2小时和6小时之间的差异比叶绿体实验中的更大。这两个细胞器的光谱都与完整叶片的光谱同步这一事实表明,这两个细胞器可能参与了细胞的O(-.)(2)爆发。在黑暗中,Ecc处理组和对照组接种Ecc 2小时和6小时后叶绿体均未显示ESR光谱,表明叶绿体中的O(-.)(2)可能起源于光合电子传递过程。