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通过分子探针和基因组稳定性分析揭示的长时间干燥后嗜球藻属CCMEE 029存活菌株的亚细胞完整性

Subcellular integrities in Chroococcidiopsis sp. CCMEE 029 survivors after prolonged desiccation revealed by molecular probes and genome stability assays.

作者信息

Billi Daniela

机构信息

Department of Biology, University of Rome Tor Vergata, Via della Ricerca Scientifica 1, 00133 Rome, Italy.

出版信息

Extremophiles. 2009 Jan;13(1):49-57. doi: 10.1007/s00792-008-0196-0. Epub 2008 Oct 18.

Abstract

Desiccation-tolerant cells must either protect their cellular components from desiccation-induced damage and/or repair it upon rewetting. Subcellular damage to the anhydrobiotic cyanobacterium Chroococcidiopsis sp. CCMEE 029 stored in the desiccated state for 4 years was evaluated at the single-cell level using fluorescent DNA strand breakage labelling, membrane integrity and potential related molecular probes, oxidant-sensing fluorochrome and redox dye. Covalent modifications of dried genomes were assessed by testing their suitability as PCR template. Results suggest that desiccation survivors avoid/and or limit genome fragmentation and genome covalent modifications, preserve intact plasma membranes and phycobiliprotein autofluorescence, exhibit spatially-reduced ROS accumulation and dehydrogenase activity upon rewetting. Damaged cells undergo genome fragmentation, loss of plasma membrane potential and integrity, phycobiliprotein bleaching, whole-cell ROS accumulation and lack respiratory activity upon rewetting. The co-occurrence of live and dead cells within dried aggregates of Chroococcidiopsis confirms that desiccation resistance is not a simple process and that subtle modifications to the cellular milieu are required to dry without dying. It rises also intriguing questions about the triggers of dead cells in response to drying. The capability of desiccation survivors to avoid and/or reduce subcellular damage, shows that protection mechanisms are relevant in the desiccation tolerance of this cyanobacterium.

摘要

耐干燥细胞必须要么保护其细胞成分免受干燥诱导的损伤,和/或在重新湿润时进行修复。使用荧光DNA链断裂标记、膜完整性和电位相关分子探针、氧化传感荧光染料和氧化还原染料,在单细胞水平上评估了干燥保存4年的嗜极蓝藻Chroococcidiopsis sp. CCMEE 029的亚细胞损伤。通过测试其作为PCR模板的适用性来评估干燥基因组的共价修饰。结果表明,干燥存活者避免和/或限制基因组片段化和基因组共价修饰,保持完整的质膜和藻胆蛋白自发荧光,在重新湿润时表现出空间上减少的活性氧积累和脱氢酶活性。受损细胞在重新湿润时会发生基因组片段化、质膜电位和完整性丧失、藻胆蛋白漂白、全细胞活性氧积累且缺乏呼吸活性。嗜极蓝藻干燥聚集体中活细胞和死细胞的共存证实,抗干燥能力不是一个简单的过程,并且需要对细胞环境进行细微的修饰才能在不死亡的情况下干燥。这也引发了关于死细胞对干燥反应触发因素的有趣问题。干燥存活者避免和/或减少亚细胞损伤的能力表明,保护机制与这种蓝藻的耐干燥性有关。

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