Micropropagation system of Anaphalis hancockii Maxim in vitro and exploration of endogenous metabolome differences coursed by plant growth regulators

Anaphalis hancockii Maxim is often used as an aromatic plant in China. To effectively protect and utilize the resources of A. hancockii , a completely rapid micropropagation system has been developed. Additionally, ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS) technology, coupled with multivariate statistical analysis and pathway enrichment analysis, was used to assess the regulatory influences of 6-benzylaminopurine (6-BA) and kinetin (KT) applied in A. hancockii . The results showed that when MS medium was supplemented with 4.0 mg/L 2,4-D and 2.0 mg/L KT, the effect of inducing callus from leaf, petiole, and root explants was the best. At this time, the leaf callus induction rate was 94.67%. Adding 3.0 mg/L KT to the medium alone had the best induction effect on adventitious shoots, and the induction rate in leaf tissue at this time was 91.96%. The optimal condition for rooting medium was MS supplement with 0.1 mg/L IBA. The use of 6-BA and KT made the endogenous metabolite content show a more up-regulated trend to a certain extent. In addition, this study found that 6-BA was advantageous to the synthesis and accumulation of phenolic acids, and KT was beneficial to the synthesis and proliferation of terpenoids. Cytokinins have a significant effect on the aromatic components of A. hancockii , and the effects of different cytokinins on aromatic components seem to be different. The results of this study indicate that plant tissue culture is an essential biotechnological means to change the pattern of aromatic components in aromatic plants.