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克菌丹对酿酒酵母诱导的坏死和凋亡性细胞死亡。

Necrotic and apoptotic cell death induced by Captan on Saccharomyces cerevisiae.

作者信息

Scariot Fernando J, Jahn Luciane, Delamare Ana Paula L, Echeverrigaray Sergio

机构信息

Institute of Biotechnology, University of Caxias do Sul, Caxias do Sul, Rio Grande Do Sul, Brazil.

Cytogene Diagnósticos Moleculares Ltda., Lajeado, Rio Grande Do Sul, Brazil.

出版信息

World J Microbiol Biotechnol. 2017 Aug;33(8):159. doi: 10.1007/s11274-017-2325-3. Epub 2017 Jul 26.

Abstract

Captan is one of the most widely used broad-spectrum fungicide applied to control several early and late diseases of grapes, apples, and other fruits and vegetables, and as other phthalimide fungicides is defined as a multisite compound with thiol-reactivity. Captan can affect non-target organisms as yeasts, modifying microbial populations and fermentation processes. In this study, we asked whether Captan thiol-reactivity and other mechanisms are involved in acute Captan-induced cell death on aerobic growing Saccharomyces cerevisiae. Thus for, we analyze cellular protein and non-protein thiols, cell membrane integrity, reactive oxygen species accumulation, phosphatidylserine externalization, and apoptotic mutants behavior. The results showed that when submitted to acute Captan treatment most cells lost their membrane integrity and died by necrosis due to Captan reaction with thiols. However, part of the cells, even maintaining their membrane integrity, lost their culture ability. These cells showed an apoptotic behavior that may be the result of non-protein thiol depletion and consequent increase of reactive oxygen species (ROS). ROS accumulation triggers a metacaspase-dependent apoptotic cascade, as shown by the higher viability of the yca1-deleted mutant. Together, necrosis and apoptosis are responsible for the high mortality detected after acute Captan treatment of aerobically growing cells of S. cerevisiae.

摘要

克菌丹是应用最为广泛的广谱杀菌剂之一,用于防治葡萄、苹果及其他果蔬的多种早期和晚期病害,与其他邻苯二甲酰亚胺类杀菌剂一样,被定义为具有硫醇反应活性的多位点化合物。克菌丹会影响非靶标生物,如酵母,改变微生物种群和发酵过程。在本研究中,我们探究了克菌丹的硫醇反应活性及其他机制是否参与其对有氧生长的酿酒酵母的急性诱导细胞死亡过程。为此,我们分析了细胞蛋白质和非蛋白质硫醇、细胞膜完整性、活性氧积累、磷脂酰丝氨酸外化以及凋亡突变体的行为。结果表明,在急性克菌丹处理下,大多数细胞因克菌丹与硫醇的反应而失去膜完整性并坏死。然而,部分细胞即使保持膜完整性,也失去了培养能力。这些细胞表现出凋亡行为,这可能是非蛋白质硫醇耗竭以及活性氧(ROS)随之增加的结果。如yca1缺失突变体具有更高的活力所示,ROS积累触发了一种依赖于metacaspase的凋亡级联反应。总之,坏死和凋亡共同导致了对有氧生长的酿酒酵母细胞进行急性克菌丹处理后检测到的高死亡率。

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