A Novel in Vitro Chemotaxis Bioassay to Assess the Response of Towards Various Test Compounds

Plant-parasitic, root-knot nematodes ( spp.) are a serious problem in agri- and horticultural crops worldwide. Understanding their complex host recognition process is essential for devising efficient and environmental-friendly management tactics. In this study, the authors report a new, simple, inexpensive, efficient, and quantitative method to analyze the chemotaxis of second-stage juveniles (J2s) using a combination of pluronic gel and agar in a petri dish. The authors quantitatively defined the concentration gradient formation of acid fuchsin on the assay plate. Using this novel assay method, the authors have accurately measured the nematode response (attraction or repulsion) to various volatile (isoamyl alcohol, 1-butanol, benzaldehyde, 2-butanone, and 1-octanol) and non-volatile (root exudates of tomato, tobacco, and marigold) compounds. Isoamyl alcohol, 1-butanol, and 2-butanone were attractive to J2s through a broad range of concentrations. On the contrary, J2s were repelled when exposed to various concentrations of 1-octanol. Despite being attractive at lower concentrations, undiluted benzaldehyde was repulsive to J2s. Tomato and tobacco root exudates were attractive to J2s while marigold root exudates repelled J2s. The present quantitative assay method could be used as a reference to screen and identify new candidate molecules that attract or repel nematodes. Plant-parasitic, root-knot nematodes ( spp.) are a serious problem in agri- and horticultural crops worldwide. Understanding their complex host recognition process is essential for devising efficient and environmental-friendly management tactics. In this study, the authors report a new, simple, inexpensive, efficient, and quantitative method to analyze the chemotaxis of second-stage juveniles (J2s) using a combination of pluronic gel and agar in a petri dish. The authors quantitatively defined the concentration gradient formation of acid fuchsin on the assay plate. Using this novel assay method, the authors have accurately measured the nematode response (attraction or repulsion) to various volatile (isoamyl alcohol, 1-butanol, benzaldehyde, 2-butanone, and 1-octanol) and non-volatile (root exudates of tomato, tobacco, and marigold) compounds. Isoamyl alcohol, 1-butanol, and 2-butanone were attractive to J2s through a broad range of concentrations. On the contrary, J2s were repelled when exposed to various concentrations of 1-octanol. Despite being attractive at lower concentrations, undiluted benzaldehyde was repulsive to J2s. Tomato and tobacco root exudates were attractive to J2s while marigold root exudates repelled J2s. The present quantitative assay method could be used as a reference to screen and identify new candidate molecules that attract or repel nematodes.