Transglutaminase activity changes during the hypersensitive reaction, a typical defense response of tobacco NN plants to TMV
Transglutaminase activity changes during the hypersensitive reaction, a typical defense response of tobacco NN plants to TMV
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DOI：
10.1111/j.1399-3054.2007.00950.x
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发表时间：
2007-10-01
影响因子：
6.4
通讯作者：
Torrigiani, Patrizia
中科院分区：
文献类型：
--
作者：
Del Duca, Stefano;Betti, Lucietta;Torrigiani, Patrizia
报道了在抗性NN烟草植物（Nicotiana tabacum L.）的过敏反应（HR）进行期间谷氨酰多胺（PA）的出现以及负责其合成的转氨酶（TGase，EC 2.3.2.13）的活性和水平的变化。CV. Samsun）对烟草花叶病毒（TMV）的抗性。用TMV或磷酸盐缓冲液（模拟）接种后0-72小时收集烟草的成熟叶。在体内合成的多胺谷氨酰衍生物（谷氨酰PAs），催化TGase活性，评价后供应标记腐胺（钚，TGase的生理底物）的叶片。结果表明，从24 h开始，单-（γ-谷氨酰）-Pu和双-（γ-谷氨酰）-Sd在TMV接种的样品中恢复，而不是在模拟接种的; 2天后，在前者，谷氨酰衍生物的量进一步增加。体外放射性测定表明，在TMV接种的叶片，TGase活性增加，从24小时起相对于模拟对照。AtPng 1 p多克隆抗体的免疫印迹分析检测到一个72 kDa的蛋白质，其量增加，在72小时TMV接种的叶片和病变丰富的地区。生物素标记的尸胺掺入试验表明，TGase活性发生在S1（含有可溶性蛋白质），S2（由细胞壁和膜释放的蛋白质）和S3（膜内在蛋白质）馏分。在S3级分，其中的变化是最相关的，TGase活性增强在模拟接种和TMV接种的样品，但刺激只持续在后一种情况下。这些数据根据TGase活性和谷氨酰多胺在防御病毒植物病原体中可能发挥的作用进行了讨论。
The occurrence of glutamyl polyamines (PAs) and changes in activity and levels of transglutaminase (TGase, EC 2.3.2.13), the enzyme responsible for their synthesis, are reported during the progression of the hypersensitive reaction (HR) of resistant NN tobacco plants (Nicotiana tabacum L. cv. Samsun) to tobacco mosaic virus (TMV). Mature leaves of tobacco were collected over 0-72 h after inoculation with TMV or phosphate buffer (mock). In vivo synthesis of polyamine glutamyl derivatives (glutamyl PAs), catalyzed by TGase activity, was evaluated after supplying labeled putrescine (Pu, a physiological substrate of TGase) to leaves. Results show that, starting from 24 h, mono-(gamma-glutamyl)-Pu and bis-(gamma-glutamyl)-Sd were recovered in TMV-inoculated samples but not in mock-inoculated ones; 2 days later, in the former, the amount of glutamyl derivatives further increased. An in vitro radiometric assay showed that, in TMV-inoculated leaves, TGase activity increased from 24 h onwards relative to mock controls. An immunoblot analysis with AtPng1p polyclonal antibody detected a 72-kDa protein whose amount increased at 72 h in TMV-inoculated leaves and in the lesion-enriched areas. A biotin-labeled cadaverine incorporation assay showed that TGase activity occurred in S1 (containing soluble proteins), S2 (proteins released by both cell walls and membranes) and S3 (membrane intrinsic proteins) fractions. In S3 fraction, where changes were the most relevant, TGase activity was enhanced in both mock-inoculated and TMV-inoculated samples, but the stimulation persisted only in the latter case. These data are discussed in the light of a possible role of TGase activity and glutamyl PAs in the defense against a viral plant pathogen.