ABSTRACT The efficacy of Taxol, a natural anticancer drug, in the treatment of various types of cancers has been certified globally. Fungal elicitors have been reported as an impressive strategy for enhancing Taxol biosynthesis. We have investigated the effect of twig age on Taxol biosynthesis and the communities of endophytic fungi. A negative correlation between Taxol content and the complexity of the endophytic fungal community in twigs was predicted. Endogenous taxoids, similar to balancing valves, might have a specific effect on controlling the microbiota assembly in Taxus twigs. Utilising the special correlation, 11 isolates of twig age‐associated fungi were used to screen new fungal elicitors involved in Taxol biosynthesis. Two efficient fungal elicitors, L01 ( Guignardia ) and J02 ( Diaporthe ), were identified, increasing the Taxol contents by 5.91‐ and 4.83‐folds, respectively. It is confirmed that effective fungal elicitors may be negatively correlated with Taxol contents in Taxus tissues. Furthermore, the J02 and L01 fungal elicitors significantly induced the jasmonic acid (JA) content, speculating the involvement of MYC2a‐controlled JA signalling in fungal elicitor‐activated Taxol biosynthesis. Our data revealed the effect of twig age on Taxol biosynthesis of Taxus and provided a novel approach to screen effective fungal elicitors involved in Taxol biosynthesis.