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过氧化氢(Hydrogen peroxide,H2O2)为活性氧(Reactive oxygen species,ROS)的一种,存在于许多生物体系中并介导植物中多种生理和生化过程。为了探讨H2O2作为信号分子在水杨酸(Salicylic acid,SA)诱导丹参培养细胞合成丹酚酸B(Salvianolic acid B,Sal B)过程中的作用,分别考察了SA和H2O2、过氧化氢酶(Catalase,CAT)、二甲基硫脲(2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide,DMTU)及咪唑(Imidazole,IMD)对苯丙氨酸解氨酶(Phenylalanine ammonia-lyase,PAL)和酪氨酸氨基转移酶(Tyrosine aminotransferase,TAT)的活性及Sal B含量的影响。结果表明,SA处理可有效地诱导丹参培养细胞中H2O2产生、PAL和TAT活性升高以及Sal B合成积累量的增加;外源施加10～30 mmol/L H2O2也可以有效促进PAL、TAT活性升高和Sal B合成积累量的增加;用H2O2的清除剂CAT处理发现,CAT对SA或外源H2O2诱导的Sal B合成积累具有消除作用,说明H2O2可能作为SA诱导Sal B积累过程中的上游信号分子起作用;用H2O2淬灭剂DMTU处理,可以有效抑制SA对Sal B合成的促进作用;用质膜烟酰胺腺嘌呤二核苷酸(Nicotinamide vadenine dinucleotide phosphate,NADPH)氧化酶(H2O2来源的主要酶)抑制剂IMD处理,可以抑制Sal B的合成,但这种抑制效果可以部分被外源施加的SA削弱,说明通过HADPH氧化酶产生的H2O2受阻时,SA诱导的Sal B合成积累也会受到抑制。表明H2O2是介导SA诱导丹参培养细胞中Sal B合成积累的信号分子。
Hydrogen peroxide (H2O2), a kind of reactive oxygen species (ROS), exists in many biological systems and mediates many physiological and biochemical processes in plants. To investigate the role of H2O2 as a signaling molecule in salicylic acid (SA) -induced salvianolic acid B (Sal B) synthesis in Salvia miltiorrhiza cells, the effects of SA and H2O2, catalase Catalase (CAT), 2- (4-carboxy-2-phenyl) -4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (DMTU) and imidazole (IMD) Effects of Phenylalanine ammonia-lyase (PAL) and Tyrosine aminotransferase (TAT) activity and Sal B content. The results showed that SA treatment could effectively induce the production of H2O2, the increase of PAL and TAT activity and the increase of the accumulation of Sal B in cultured Salvia miltiorrhiza cells. Exogenous application of 10-30 mmol / L H2O2 also effectively promoted the activity of PAL and TAT H2O2 scavenging agent CAT treatment showed that CAT inhibited the accumulation of Sal B induced by SA or exogenous H2O2, indicating that H2O2 may serve as an upstream signal during SA-induced Sal B accumulation Molecules play a role; with H2O2 quencher DMTU treatment, SA can effectively inhibit the synthesis of Sal B role; plasma membrane nicotinamide adenine dinucleotide (Nicotinamide vadenine dinucleotide phosphate, NADPH oxidase (H2O2 source of the main Enzyme) inhibitor IMD could inhibit the synthesis of Sal B, but this inhibitory effect could be partially attenuated by exogenously applied SA, indicating that accumulation of SA-induced Sal B by the HADPH oxidase was blocked when it was blocked inhibition. H2O2 is a signaling molecule that mediates SA-induced accumulation of Sal B in SA-cultured Salvia miltiorrhiza cells.