Soybean cyst nematodes (SCNs) are a significant disease that causes yield loss and reducing seed quality in soybeans (Glycine max). Developing SCN-resistant soybean varieties can minimize the need for insecticide use and reduce yield loss. Cinnamate-4-hydroxylase (C4H) and laccase (Lac) are key enzymes in the lignin synthesis pathway. In this study, SCN stress significantly promoted lignin accumulation in soybean roots and upregulated the expression of lignin signaling pathway genes GmC4H (Glyma.02G236500), GmLac55 (Glyma.13G076900), and GmLac85 (Glyma.20G051900). Using Agrobacterium rhizogenes-mediated transformation, the pNI900 expression vector was introduced into the soybean cultivar Williams 82 to generate GmLac55-overexpressing plants. The overexpression of GmLac55 enhanced soybean roots resistance to SCN and inhibited the further development of J2 larvae. Our study presents a strategy for increasing SCN resistance in soybean through Agrobacterium-mediated targeted mutagenesis of the GmLac55 gene.

Soybean cyst nematode (SCN) development depends on syncytium formation, which requires cell-wall degradation and fusion. Lignin, the main barrier in cell walls, is critical for SCN resistance. 4-Coumarate: CoA ligase (4CL) drives the phenylpropanoid pathway by converting p-coumaric acid to p-coumaroyl-CoA, supplying lignin precursors. Here, resistant cv. Huipizhiheidou accumulated more lignin than susceptible Williams 82 after SCN inoculation. SCN stress induced distinct Gm4CL-family expression profiles across cultivars; Gm4CL3 and Gm4CL4 were markedly upregulated in Huipizhiheidou. Transient expression of Gm4CL3 in tobacco thickened leaf cell walls, implying enhanced wall reinforcement against SCN. Thus, 4CLs, especially Gm4CL3, may promote lignin deposition and secondary wall thickening to strengthen soybean SCN resistance.