Progressive Vertical and Horizontal Phytocompound Changes during Agarwood Formation in Aquilaria sinensis after Geotrichum candidum Injection.

(1) Background: Agarwood is an aromatic resin produced by the host tree through an immunological response against biotic and abiotic stress. The aim was, first, to use the fungus Geotrichum candidum to stimulate compound changes in Aquilaria sinensis horizontally (color formation) and vertically (cutting layers) after injection with it. (2) Methods: Horizontal and vertical sections were collected and separated five months after injection with the fungal broth. Two grams of dry powder was mixed with 20 mL methanol for 3 h at room temperature, and the solution was vibrated in an ultrasonic cleaner bath at 40 °C for 1 h. After vacuum drying, a concentration of 10 mg/mL of the tested samples in methanol was prepared for reversed-phase high-performance liquid chromatography (RP-HPLC), gas chromatography/mass spectrometry (GC/MS), and thin-layer chromatography (TLC) analysis. (3) Results: The horizontal changes in the compounds and their concentrations were associated with color. Compared to the normal (N) group, G. candidum injection stimulated more compounds at RT 27-42 in the white (W) group, brown (BR) group, and black (B) group. Furthermore, a significant increase in fatty acids was observed in the W group, implying an early plant response after G. candidum injection. In the BR group, the compounds were more similar to commercial agarwood (Out group). In the B group, alkaloids were the main compounds. Vertical changes in the main compounds were not observed, although the compound level varied. A TLC analysis determined the main compounds in the BR group at 254 nm and in the B group at 365 nm. Higher fatty acid levels were found in L6 and L5 and were correlated with higher terpenoid and sesquiterpene levels, suggesting that these compounds were possibly the first stage of agarwood formation. A GC/MS analysis demonstrated that the main compound groups were almost identical to the BR parts. (4) Conclusions: The injection of G. candidum led A. sinensis to synthesize different phytochemicals horizontally, not vertically, in the BR group.