Nitric oxide regulates shikonin formation in suspension-cultured Onosma paniculatum cells.
Nitric oxide regulates shikonin formation in suspension-cultured Onosma paniculatum cells.
复制标题
DOI：
10.1093/pcp/pcn178
复制
发表时间：
2009
影响因子：
4.9
通讯作者：
Shu-jing Wu;J. Qi;Wen-Ju Zhang;Shao-hua Liu;F. Xiao;Ming-sheng Zhang;Guo-Hua Xu;Wei-guang Zhao-Wei-guang-Z
中科院分区：
文献类型：
--
作者：
Shu-jing Wu;J. Qi;Wen-Ju Zhang;Shao-hua Liu;F. Xiao;Ming-sheng Zhang;Guo-Hua Xu;Wei-guang Zhao-Wei-guang-Z
内源性一氧化氮（NO）参与调控滇紫草（Onosma paniculatum）细胞中紫草素的形成。细胞接种到紫草素生产培养基后产生的NO在培养2天后达到最高水平，是实验开始时的16倍，并在6天内维持较高水平。一氧化氮合酶（NOS）抑制剂N(ω)-硝基-L-精氨酸（L - NNA）和硝酸还原酶（NR）抑制剂叠氮化钠（SoA），因其抑制NO的生物合成，显著降低了紫草素的形成。这种降低可通过硝普钠（SNP）提供的外源NO得到缓解甚至消除，这表明NO生物合成的抑制导致紫草素形成减少。然而，当由禾谷丝核菌（Rhizoctonia cerealis）激发子上调内源性NO生物合成时，紫草素的产量进一步提高，显示出对激发子诱导的NO爆发的依赖性。实时荧光定量PCR分析表明，NO可显著上调编码参与紫草素生物合成关键酶的PAL、PGT和HMGR的表达。这些结果表明，NO在滇紫草细胞中紫草素的形成过程中起着关键作用。
Endogenously occurring nitric oxide (NO) is involved in the regulation of shikonin formation in Onosma paniculatum cells. NO generated after cells were inoculated into shikonin production medium reached the highest level after 2 d of culture, which was 16 times that at the beginning of the experiment, and maintained a high level for 6 d. A nitric oxide synthase (NOS) inhibitor, N(omega)-nitro-L-arginine (L-NNA), and a nitrate reductase (NR) inhibitor, sodium azide (SoA), consistent with their inhibition of NO biosynthesis, decreased shikonin formation significantly. This reduction could be alleviated or even abolished by exogenous NO supplied by sodium nitroprusside (SNP), suggesting that the inhibition of NO biosynthesis resulted in decreased shikonin formation. However, when endogenous NO biosynthesis was up-regulated by the elicitor from Rhizoctonia cerealis, shikonin production was enhanced further, showing a dependence on the elicitor-induced NO burst. Real-time PCR analysis showed that NO could significantly up-regulate the expression of PAL, PGT and HMGR, which encode key enzymes involved in shikonin biosynthesis. These results demonstrated that NO plays a critical role in shikonin formation in O. paniculatum cells.