Involvement of nitric oxide in ultraviolet B-induced activation of phenylalanine ammoniumlyase and stimulation of flavonoid biosynthesis in Ginkgo biloba leaves

Previous studies have shown that ultraviolet B (UV-B) irradiation can stimulate biosynthesis of secondary metabolites. Several other studies have demonstrated that nitric oxide (NO) is an important signal molecule that plays a key role in plant defence responses to UV-B irradiation. However, the signalling mechanism of NO involvement in UV-B-induced secondary-metabolite synthesis is unclear. The purpose of the present study is to investigate the role of NO in the accumulation UV-B-induced secondary metabolite in the leaves of Ginkgo biloba. Our results showed that UV-B irradiation induced multiple biological responses in the leaves of G. biloba, including increases in both NO production and nitric oxide synthase (NOS) activity, and subsequent activation of phenylalanine ammoniumlyase (PAL) and synthesis of flavonoids. Application of NO via the donor sodium nitroprusside (SNP) enhanced UV-B-induced PAL activity and increased accumulation of flavonoids in G. biloba leaves. Both the NOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) and the NO scavenger 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (c-PTIO) reduced the production of NO. Moreover, UV-B-induced increase in PAL activity and flavonoid accumulation were suppressed by L-NAME and c-PTIO. These findings suggest a causal relationship between NO release and both PAL activity and flavonoid accumulation under UV-B irradiation. In addition, the results also indicate that NO, produced via NOS-like activity in ginkgo leaves subjected to UV-B irradiation, might act as an essential signal molecule for triggering the activation of PAL and synthesis of flavonoids.