水杨酸诱发的NO介导了丹参悬浮培养细胞中丹酚酸B的生物合成

Nitric Oxide Triggered by Salicylic Acid Mediates the Biosynthesis of Salvianolic Acid B in Salvia miltiorrhiza Suspension Cell Culture

水杨酸(SA)可诱导丹参悬浮培养细胞中一氧化氮(NO)产生、苯丙氨酸解氨酶(PAL)活化及丹酚酸B(Sal B)的生物合成。为了阐明NO对丹参悬浮培养细胞中Sal B生物合成的影响及作用机理,本实验利用NO供体硝普钠(SNP)、NO合成酶抑制剂L-NNA(Nω-nitro-L-arginine)、NO淬灭剂c PITO(carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide)以及PAL抑制剂L-AOPP(L-2-aminooxygen-3-phenyl acrylic acid)分别处理丹参悬浮培养细胞,并对其胞内NO水平、PAL活性和Sal B积累量进行了检测。结果表明,硝普钠(SNP)处理显著促进了NO产生、PAL活性和Sal B的积累,而L-NNA和c PITO抑制上述过程,说明NO诱发PAL活性提高并参与了SA诱导的Sal B生物合成;L-AOPP显著抑制了PAL活性及Sal B积累,却对NO产生没有显著影响,揭示NO位于PAL的上游。这说明SA诱发的NO产生、PAL活化及Sal B合成之间存在因果关系,即NO通过激活PAL触发Sal B生物合成。

Salicylic acid（SA） induced nitric oxide（NO） generation,Phenylalanine ammonialyase（PAL） activation,and salvianolic acid B（Sal B） biosynthesis. To determine the role of NO in SA-induced Sal B biosynthesis,the effects of NO donor sodium nitroprusside（SNP）,NO synthase inhibitor L-NNA（Nω-nitro-L-arginine）,NO scavenger carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide（c PITO）,and PAL inhibitor L-AOPP（L-2-aminooxygen-3-phenyl acrylic acid） on SA-induced NO generation,PAL activation,and Sal B accumulation were stud ied ind ivid ually. Pretreatment of the cells with SNP increased SA-ind uced NO g eneration,PAL activation and Sal B accumulation,which sug g ested that NO activated PAL and was involved in SA-ind uced Sal B b iosynthesis. L-AOPP sup p ressed PAL activity and Sal B accumulation,b ut d id not affect SA-ind uced NO g eneration,ind icating that NO acted as an up stream sig nal of PAL. Results ind icated that there was a causal relationship between SA-induced NO generation, PAL activation, and Sal B biosynthesis in Salvia miltiorrhiza suspension cell culture. Via activation of PAL,NO mediated the SA-induced Sal B biosynthesis.