Nematotoxic coumarins from Angelica pubescens Maxim. f. biserrata Shan et Yuan roots and their physiological effects on Bursaphelenchus xylophilus

The ethanol extracts from the roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan was toxic against the pine wood nematode Bursaphelenchus xylophilus. The ethyl acetate-soluble fraction derived from this extract increased its potency with a mortality of 95.25% in 72 hr at 1.0 mg/mL. Four nematotoxic coumarins were obtained from the ethyl acetate extract by bioassay-guided isolation. These were identified as osthole 1 , columbianadin 2 , bergapten 3 and xanthotoxin 4 by mass and nuclear magnetic resonance spectral data analysis. The LC₅₀ values against B. xylophilus in 72 hr were 489.17, 406.74, 430.08, and 435.66 μM, respectively. These compounds also altered the smooth morphology of the B. xylophilus exoskeleton to a rough and pitted appearance as visualized by electron microscopy. The coumarins 1-4 possessed significant acetylcholinesterase inhibitory activities but had negligible effects on amylase and cellulase. This research provides additional clues to the nematotoxic mechanism of coumarins against the pine wood nematode B. xylophilus. This work will assist in the development of coumarin nematicides with enhanced activity using molecular modifications of the core coumarin structure.