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表没食子儿茶素没食子酸酯(EGCG)胁迫下铜绿微囊藻的生理及基因表达变化

Changes in the physiology and gene expression of Microcystis aeruginosa under EGCG stress.

作者信息

Lu Yaping, Wang Jin, Yu Yang, Shi Limei, Kong Fanxiang

机构信息

Biological Experiment Teaching Center, College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China; State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008, China.

Biological Experiment Teaching Center, College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Chemosphere. 2014 Dec;117:164-9. doi: 10.1016/j.chemosphere.2014.06.040. Epub 2014 Jul 10.

Abstract

EGCG (Epigallocatechin-3-gallate) has an allelopathic inhibitory effect on Microcystis aeruginosa. Cellular structure, physiological and biochemical reactions and gene expression were examined to explore the mechanism of inhibition. As was shown in electron microscopy, the structure of the cell wall, cell membrane and thylakoid was disrupted by EGCG. EGCG also reduced the efficiency of photosynthesis and the electron transfer rate in M. aeruginosa cells, as was determined with a flow cytometer. Quantitative real-time PCR analysis demonstrated that gene expression of the core proteins of the photosynthesis centers PSI and PSII and ATP synthase were reduced, while the expression of the phycobilisome degradation protein A gene (nbl A) was elevated. The expression of the universal stress protein gene increased, which would enhance the adaptive capacity of Microcystis cells to polyphenols and oxidative stress. Furthermore, EGCG elevated the level of reactive oxygen species (ROS) in M. aeruginosa cells, and thus caused oxidative cellular damage. When treated with EGCG at low concentrations (10 and 40 mg L(-)(1)), the cells were able to activate defense systems to degrade the excess ROS. But at a concentration of 70 mg L(-)(1), oxidative stress exceeded tolerance limits, and the cells were severely damaged. We concluded that damage to photosynthesis and oxidative stress were the primary mechanisms for the allelopathic effect of EGCG on M. aeruginosa.

摘要

表没食子儿茶素没食子酸酯(EGCG)对铜绿微囊藻具有化感抑制作用。通过检测细胞结构、生理生化反应及基因表达来探究其抑制机制。电子显微镜观察结果显示,EGCG破坏了铜绿微囊藻的细胞壁、细胞膜及类囊体结构。流式细胞仪检测结果表明,EGCG还降低了铜绿微囊藻细胞的光合作用效率及电子传递速率。定量实时PCR分析表明,光合作用中心PSI和PSII的核心蛋白以及ATP合酶的基因表达降低,而藻胆体降解蛋白A基因(nbl A)的表达升高。普遍应激蛋白基因的表达增加,这会增强微囊藻细胞对多酚和氧化应激的适应能力。此外,EGCG提高了铜绿微囊藻细胞内活性氧(ROS)水平,从而导致细胞氧化损伤。当用低浓度(10和40 mg L⁻¹)的EGCG处理时,细胞能够激活防御系统来降解过量的ROS。但在浓度为70 mg L⁻¹时,氧化应激超过耐受极限,细胞受到严重损伤。我们得出结论,光合作用受损和氧化应激是EGCG对铜绿微囊藻化感作用的主要机制。

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