Optimization and induction effect evaluation of complex inducer of Aquilaria sinensis based on factorial design

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Agarwood is a kind of valuable traditional Chinese medicine. However, natural agarwood has a long formation cycle and is in short supply. Herein, the complex inducer of artificial agarwood was screened and optimised based on design of experiments (DoE). Single factor design experiments were conducted to evaluate the effects of different concentrations of methyl jasmonate (A-MeJA), formic acid (B-FA) and Botryosphaeria rhodina A13(C-A13) on the in vitro branches of A. sinensis at different induction times (D-Time). The 2 k factorial design was used to further optimise the screening of the complex inducer. Single factor experiments determined the concentration of the inducers acting on the branches of A. sinensis . The order of the four factors obtained by 2 k factorial experiment was D-Time > A-MeJA > B-FA > C-A13. The optimal complex inducer was 1%MeJA + 1%FA + A13. Verification through an in vivo field experiment of 18 batches of A. sinensis , artificial agarwood met the requirements of the Pharmacopoeia of People’s Republic of China 2020 (ChP 2020), an indication that the complex inducer could efficiently promote the formation of agarwood (in 9 months). These findings provide essential data and new ideas for the efficient induction of agarwood and the production of high-quality substitutes for natural agarwood.