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首页> 美国卫生研究院文献>Plant Physiology >Use of a New Tetrazolium-Based Assay to Study the Production of Superoxide Radicals by Tobacco Cell Cultures Challenged with Avirulent Zoospores of Phytophthora parasitica var nicotianae
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Use of a New Tetrazolium-Based Assay to Study the Production of Superoxide Radicals by Tobacco Cell Cultures Challenged with Avirulent Zoospores of Phytophthora parasitica var nicotianae

机译：使用新的基于四氮唑的检测方法研究 烟草细胞培养物对无毒物质的挑战超氧自由基 寄生疫霉菌的游动孢子 烟草科

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The relationship between the production of reactive oxygen species and the hypersensitive response (HR) of tobacco (Nicotiana tabacum L.) toward an incompatible race of the Oomycete Phytophthora parasitica var nicotianae has been investigated. A new assay for superoxide radical (O2−) production based on reduction of the tetrazolium dye sodium,3′-(1-[phenylamino-carbonyl]-3,4-tetrazolium)-bis(4-methoxy-6-nitro) benzene-sulfonic acid hydrate (XTT) has enabled the quantitative estimation of perhydroxyl/superoxide radical acid-base pair (HO2·/O2−) production during the resistant response. Tobacco suspension cells were inoculated with zoospores from compatible or incompatible races of the pathogen. Subsequent HO2·/O2− production was monitored by following the formation of XTT formazan. In the incompatible interaction only, HO2·/O2− was produced in a minor burst between 0 and 2 h and then in a major burst between 8 and 10 h postinoculation. During this second burst, rates of XTT reduction equivalent to a radical flux of 9.9 × 10−15 mol min−1 cell−1 were observed. The HO2·/O2− scavengers O2− dismutase and Mn(III)desferal each inhibited dye reduction. An HR was observed in challenged, resistant cells immediately following the second burst of radical production. Both scavengers inhibited the HR when added prior to the occurrence of either radical burst, indicating that O2− production is a necessary precursor to the HR.
机译：研究了活性氧物质的产生与烟草（Oniccete tabacum L.）对卵形疫霉（Phytophthora parasitica var nicotianae）不相容小种的超敏反应（HR）之间的关系。基于还原四唑染料钠3'-（1- [1- [苯基氨基-羰基] -3,4-四唑]-双（4-）的超氧自由基（O2 -）产生的新测定法甲氧基-6-硝基）苯磺酸水合物（XTT）能够定量估计过羟基/超氧化物基酸对（HO2 · / O2 -）的产量在抵抗反应期间。用来自病原体相容或不相容种族的游动孢子接种烟草悬浮细胞。通过跟踪XTT甲maz的形成来监测随后的HO2 · / O2 -的产生。仅在不兼容的互动中， HO2 · / O2 -原为 在0到2小时之间以小幅爆发产生，然后在 接种后8至10小时爆发。在这第二秒 爆发，XTT降低的速率等于9.9的基本通量 ×10 −15 mol min −1 单元格 -1 是 观测到的。的 HO2 · / O2 -清除剂 O2 -歧化酶和Mn（III）延迟 抑制染料还原。在挑战，抵抗力中观察到HR 自由基产生的第二次爆发后立即产生细胞。 两种清除剂均在HR发生之前添加时抑制了HR。 任一基团爆裂，表明O2 - 生产是人力资源的必要前提。

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