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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Reactive oxygen and nitrogen (ROS and RNS) species generation and cell death in tomato suspension cultures-Botrytis cinerea interaction
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Reactive oxygen and nitrogen (ROS and RNS) species generation and cell death in tomato suspension cultures-Botrytis cinerea interaction

机译：番茄悬浮培养物-灰葡萄孢互作中活性氧和氮（ROS和RNS）的产生和细胞死亡

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This article reports events connected to cell survival and Botrytis cinerea infection development in cell suspension cultures of two tomato cultivars which show different levels of susceptibility to the pathogen: cv. Corindo (more susceptible) and cv. Perkoz (less susceptible). In parallel changes in reactive oxygen (ROS) and nitrogen (RNS) species generation and in S-nitrosoglutathione reductase (GSNOR) activity were studied. In vivo staining methods with acridine orange (AO) and ethidium bromide (EB) as well as fluorescent microscopy were used to assess tomato and B. cinerea cells death. The biochemical studies of ROS and RNS concentrations in plant cell extract were complemented by in vivo ROS and nitric oxide (NO) imaging using nitro blue tetrazolium (NBT), diaminobenzidine (DAB) and diaminofluorescein diacetate (DAF-DA) staining methods, and confocal microscope technique. B. cinerea infection proceeded slower in Perkoz cell cultures. It was evidenced by measuring the pathogen conidia germination and germination tube development in which nuclei revealing cell death dominated. Two different types of tomato cell death were observed: cells with necrotic nuclei dominated in Corindo whereas in Perkoz cells with characteristic of vacuolar death type prevailed. In Perkoz cells, constitutive levels of NO and S-nitrosothiols (SNO) were significantly higher and hydrogen peroxide (H2O2) and superoxide anion (O-2 (-)) concentrations were slightly higher as compared with Corindo cells. Moreover, increases in these molecule concentrations as a result of B. cinerea inoculation were observed in both, Perkoz and Corindo cell cultures. The enzymatic GSNOR activity seems to be an important player in controlling the SNO level in tomato cells. Involvements of the studied compounds in molecular mechanisms of tomato resistance to B. cinerea are discussed in the paper.
机译：本文报道了两个番茄品种的细胞悬浮培养中与细胞存活和灰葡萄孢感染有关的事件，这些品种对病原体的敏感性不同。 Corindo（更容易受到感染）和简历。 Perkoz（较不敏感）。同时研究了活性氧（ROS）和氮（RNS）物种生成以及S-亚硝基谷胱甘肽还原酶（GSNOR）活性的变化。使用with啶橙（AO）和溴化乙锭（EB）的体内染色方法以及荧光显微镜检查来评估番茄和灰质芽孢杆菌的死亡。植物细胞提取物中ROS和RNS浓度的生化研究通过使用硝基蓝四唑（NBT），二氨基联苯胺（DAB）和二氨基荧光素二乙酸酯（DAF-DA）染色方法以及共聚焦的体内ROS和一氧化氮（NO）成像进行补充显微镜技术。在Perkoz细胞培养中，灰质芽孢杆菌的感染进行得较慢。通过测量病原体分生孢子萌发和发芽管发育（其中显示细胞死亡的细胞核占主导）可以证明这一点。观察到两种不同类型的番茄细胞死亡：在Corindo中具有坏死核的细胞占主导，而在具有液泡死亡类型特征的Perkoz细胞中占优势。与Corindo细胞相比，在Perkoz细胞中，NO和S-亚硝基硫醇（SNO）的组成水平显着较高，过氧化氢（H2O2）和超氧阴离子（O-2（-））的浓度略高。此外，在Perkoz和Corindo细胞培养物中均观察到由于灰质芽孢杆菌接种而导致这些分子浓度增加。酶促GSNOR活性似乎是控制番茄细胞中SNO水平的重要参与者。本文讨论了所研究化合物对番茄对灰葡萄孢的抗性的分子机制。

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