Bioassay-Guided Isolation of Nematicidal Artemisinic Acid and Dihydroartemisinic Acid from Artemisia annua L. and Evaluation of Their Activity against Meloidogyne incognita.

The extract of Artemisia annua L. has been shown to possess the nematicidal activity but the potent constituents were unclear. Herein, two nematicidal sesquiterpenoid acids artemisinic acid (ArA) and dihydroartemisinic acid (DHArA) were isolated from the methanol extract of the aerial parts of A. annua L. by the bioassay-guided isolation method using Meloidogyne incognita second stage juveniles (J2s) as the screening target. The in vitro activity, control efficacy in the pot experiment, and toxic effects of these two natural compounds against M. incognita were evaluated. The in vitro results showed that the EC 50/48h values of ArA and DHArA were 0.37 mM and 0.76 mM against J2s, respectively. In the pot experiment, ArA and DHArA at the dose of 5 mg (a.i.)/pot could achieve the same level of control efficacy compared with avermectins at 2 mg (a.i.)/pot. Microscopic observations indicated the obvious toxic symptoms of J2s after ArA- and DHArA-treatment, including the shrinking body, imperfect body wall, and undiscerning organs. The physiological and biochemical studies, together with the toxic symptoms, revealed that ArA and DHArA had great impacts on the membrane system of J2s. Additionally, ArA occurring the α, β-unsaturated carbonyl was demonstrated to be reactive with glutathione (GSH) and cause the decrease of GSH content in J2s. Taken together, the present study suggests that ArA and DHArA or ArA- and DHArA-based extracts of A. annua L. have a substantial potential to be used as botanical agents for integrated disease management programs against root-knot nematodes in crop protection.