The most economically important centellosides, madecassoside and asiaticoside, are produced in very low amounts in Centella asiatica (L.) Urban roots. With the aim of overcoming this limitation, in this work we compared the effect of different elicitors on centelloside production in hairy root cultures established by Agrobacterium rhizogenes infection. Additionally, elicitor-induced changes in the expression of key biosynthetic genes were studied to shed light on the regulation of the triterpene metabolic pathway. The elicitors tested were coronatine and methyl jasmonate, added separately or together, and salicylic acid. The content of the four main centellosides (asiaticoside, madecassoside, asiatic acid and madecassic acid) was determined by HPLC/DAD, and the expression level of genes after elicitation was analyzed by real-time quantitative polymerase chain reaction. The greatest increase in the production of centellosides, especially madecassoside, was achieved with coronatine, applied alone or with methyl jasmonate. This treatment also enhanced the expression of the target genes, particularly at the start of elicitation. By far the most highly expressed were those involved in oxidations, indicating that the tested elicitors did not act specifically on key genes in the centelloside biosynthetic pathway. These results support that hairy roots are a promising biotechnological platform for improved centelloside production and that this approach warrants further research.

经济上最重要的积雪草苷、羟基积雪草苷和积雪草苷在积雪草(L.) Urban 根中的产量非常低。为了克服这一限制，在这项工作中，我们比较了不同诱导剂对发根农杆菌建立的毛状根培养物中积雪草苷产生的影响感染。此外，研究了诱导剂诱导的关键生物合成基因表达的变化，以阐明三萜代谢途径的调节。测试的诱导剂是分别或一起添加的coronatine和茉莉酸甲酯，以及水杨酸。采用HPLC/DAD测定四种主要积雪草苷（积雪草苷、羟基积雪草苷、积雪草酸和羟基积雪草酸）的含量，并通过实时定量聚合酶链反应分析诱导后基因的表达水平。使用coronatine，单独使用或与茉莉酸甲酯一起使用时，积雪草苷的产量增加最大，尤其是羟基积雪草苷。这种处理还增强了靶基因的表达，特别是在诱导开始时。到目前为止，表达最高的是参与氧化的那些，表明测试的诱导子没有特异性地作用于积雪草苷生物合成途径中的关键基因。这些结果支持毛状根是一种有前途的生物技术平台，可用于改善积雪草苷的生产，并且这种方法值得进一步研究。