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盐处理的鸭茅悬浮培养体细胞胚胎发生过程中小分子巯基和二硫键的氧化还原状态

Redox state of low-molecular-weight thiols and disulphides during somatic embryogenesis of salt-treated suspension cultures of Dactylis glomerata L.

机构信息

Department of Biochemistry, Sofia University, Sofia, Bulgaria.

出版信息

Free Radic Res. 2012 May;46(5):656-64. doi: 10.3109/10715762.2012.667565. Epub 2012 Mar 15.

Abstract

The tripeptide antioxidant γ-L-glutamyl-L-cysteinyl-glycine, or glutathione (GSH), serves a central role in ROS scavenging and oxidative signalling. Here, GSH, glutathione disulphide (GSSG), and other low-molecular-weight (LMW) thiols and their corresponding disulphides were studied in embryogenic suspension cultures of Dactylis glomerata L. subjected to moderate (0.085 M NaCl) or severe (0.17 M NaCl) salt stress. Total glutathione (GSH + GSSG) concentrations and redox state were associated with growth and development in control cultures and in moderately salt-stressed cultures and were affected by severe salt stress. The redox state of the cystine (CySS)/2 cysteine (Cys) redox couple was also affected by developmental stage and salt stress. The glutathione half-cell reduction potential (E(GSSG/2 GSH)) increased with the duration of culturing and peaked when somatic embryos were formed, as did the half-cell reduction potential of the CySS/2 Cys redox couple (E(CySS/2 Cys)). The most noticeable relationship between cellular redox state and developmental state was found when all LMW thiols and disulphides present were mathematically combined into a 'thiol-disulphide redox environment' (E(thiol-disulphide)), whereby reducing conditions accompanied proliferation, resulting in the formation of pro-embryogenic masses (PEMs), and oxidizing conditions accompanied differentiation, resulting in the formation of somatic embryos. The comparatively high contribution of E(CySS/2 Cys) to E(thiol-disulphide) in cultures exposed to severe salt stress suggests that Cys and CySS may be important intracellular redox regulators with a potential role in stress signalling.

摘要

三肽抗氧化剂 γ-L-谷氨酰-L-半胱氨酰-甘氨酸,或谷胱甘肽(GSH),在清除活性氧(ROS)和氧化信号中起核心作用。在这里,研究了处于胚胎发生悬浮培养中的 Dactylis glomerata L.中的谷胱甘肽(GSH)、谷胱甘肽二硫化物(GSSG)和其他低分子量(LMW)硫醇及其相应的二硫化物,这些物质受到中度(0.085 M NaCl)或重度(0.17 M NaCl)盐胁迫的影响。在对照培养物和中度盐胁迫培养物中,总谷胱甘肽(GSH+GSSG)浓度和氧化还原状态与生长和发育有关,并且受到严重盐胁迫的影响。半胱氨酸(CySS)/2 半胱氨酸(Cys)氧化还原对的氧化还原状态也受到发育阶段和盐胁迫的影响。谷胱甘肽半细胞还原电势(E(GSSG/2 GSH))随着培养时间的延长而增加,当体细胞胚胎形成时达到峰值,CySS/2 Cys 氧化还原对的半细胞还原电势(E(CySS/2 Cys))也是如此。在所有 LMW 硫醇和二硫化物都存在的情况下,将它们数学组合成一个“硫醇-二硫化物氧化还原环境”(E(thiol-disulphide))时,细胞氧化还原状态与发育状态之间发现了最明显的关系,在这种情况下,还原条件伴随着增殖,导致形成原胚性细胞团(PEMs),而氧化条件伴随着分化,导致形成体细胞胚胎。与严重盐胁迫下的培养物相比,E(CySS/2 Cys)对 E(thiol-disulphide)的贡献相对较高,这表明 Cys 和 CySS 可能是细胞内重要的氧化还原调节剂,在应激信号中可能具有潜在作用。

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