Eutrema japonicum is a perennial herb belonging to the Eutrema genus in the crucifer family. In recent years, numerous substances with notable pharmacological activities have been successfully isolated from E. japonicum. Despite significant advancements in related research, the efficacy of the rhizome extract of E. japonicum against root-knot nematodes remains unknown. In this study, the rhizome extract of E. japonicum was used as raw material to demonstrate the inhibitory and nematicidal effects of the extract on Meloidogyne enterolobii. The results showed that the LC₅₀ of the E. japonicum rhizome extract on second-stage juveniles (J2s) was 69.590 mg/mL and 22.336 mg/mL at 24 h and 48 h after treatment, respectively. The mortality rate of J2s reached 88.93% at 48 h post-treatment when the concentration was 200 mg/mL, and the inhibition rate of single-egg hatching reached 88.14%. This study analyzed the chemical composition of the ethanol extract of E. japonicum, and 10 organosulfur compounds and lipid compounds with insecticidal and antibacterial effects were preliminarily screened out. Among them, sec-butyl isothiocyanate and geraniol were further investigated for their nematicidal activity, demonstrating high efficacy against M. enterolobii. Moreover, we conducted network pharmacology analysis and RT-qPCR analysis to predict the potential inhibitory mechanisms of sec-butyl isothiocyanate and geraniol on M. enterolobii. These findings offer a scientific foundation and theoretical framework for utilizing E.japonicum as a potential raw material for developing novel natural plant nematicides.

Meloidogyne javanica is one of the most widespread and economically important sedentary endoparasites. In this study, a comparative transcriptome analysis of M. javanica between pre-parasitic second-stage juveniles (Pre-J2) and parasitic juveniles (Par-J3/J4) was conducted. A total of 48,698 unigenes were obtained, of which 18,826 genes showed significant differences in expression (p < 0.05). In the differentially expressed genes (DEGs) from transcriptome data at Par-J3/J4 and Pre-J2, a large number of unigenes were annotated to the C-type lectin (CTL, Mg01965), the cathepsin L-like protease (Mi-cpl-1), the venom allergen-like protein (Mi-mps-1), Map-1 and the cellulase (endo-β-1,4-glucanase). Among seven types of lectins found in the DEGs, there were 10 CTLs. The regulatory roles of Mj-CTL-1, Mj-CTL-2 and Mj-CTL-3 in plant immune responses involved in the parasitism of M. javanica were investigated. The results revealed that Mj-CTL-2 could suppress programmed cell death (PCD) triggered by Gpa2/RBP-1 and inhibit the flg22-stimulated ROS burst. In situ hybridization and developmental expression analyses showed that Mj-CTL-2 was specifically expressed in the subventral gland of M. javanica, and its expression was up-regulated at Pre-J2 of the nematode. In addition, in planta silencing of Mj-CTL-2 substantially increased the plant resistance to M. javanica. Moreover, yeast co-transformation and bimolecular fluorescence complementation assay showed that Mj-CTL-2 specifically interacted with the Solanum lycopersicum catalase, SlCAT2. It was demonstrated that M. javanica could suppress the innate immunity of plants through the peroxide system, thereby promoting parasitism.