GmSRC7 is a broad-spectrum antiviral R gene from soybean, but its downstream and functionally related genes remain unclear. Virus-induced gene silencing (VIGS) assays in Nicotiana benthamiana (Nb) showed that suppression of several gene families—WRKY transcription factors, chaperones, ethylene pathway components, MAPK cascade elements, salicylic acid (SA) signaling genes, calcium-dependent protein kinases, nuclear migration proteins, RNA replication-related genes, and immune regulators—consistently weakened GmSRC7-mediated resistance to Soybean Mosaic Virus (SMV) and Tobacco Mosaic Virus (TMV). Targeted silencing of four regulatory genes—NbEDS1, NbARF1, NbSGT1, and NbCOI1—markedly enhanced GmSRC7-mediated resistance to SMV and TMV in our experiments. Silencing the serine/threonine kinase gene NbPBS1 increased GmSRC7-conferred resistance to SMV but did not significantly alter its resistance to TMV. Transient expression assays showed that NbARF1, NbSGT1, and NbCOI1 antagonize GmSRC7-mediated defense against SMV and TMV, whereas NbPBS1 specifically suppresses anti-SMV activity without affecting TMV resistance. Transient overexpression of SA-degrading enzymes (AtS3H, AtS5H, and NahG) significantly reduced GmSRC7-conferred resistance to SMV, indicating that SA is essential for this R protein-mediated defense. Genes were also grouped by immune pathways and function: co-expression of chaperone family genes inhibited GmSRC7 activity against SMV and TMV, while co-expression of WRKY family genes enhanced anti-SMV activity of GmSRC7. Finally, transient silencing of soybean genes GmEDS1, GmSGT1-1, GmSGT1-2, GmJAR1, and GmSGS3 compromised GmSRC7-mediated resistance to SMV.

Populus hopeiensis is an important native poplar species in northern China, but herbivorous insect damage seriously affects its growth and ecological function. WRKY transcription factors play important roles in plant stress responses; however, the function of WRKY23 homologs in woody plant resistance to chewing herbivores remains unclear. In this study, a herbivore-responsive WRKY transcription factor gene, PhWRKY23, was identified from P. hopeiensis. PhWRKY23 expression was significantly induced by Spodoptera litura feeding, with a maximum increase of approximately 69.71-fold the control level at the highest damage level, and the encoded protein was predominantly localized in the nucleus. To investigate its function, PhWRKY23-overexpressing and RNA interference transgenic lines were generated. In the choice feeding assay, the consumed leaf area of PhWRKY23-overexpressing plants was approximately 85.5% lower than that of WT plants after 8 h. In the no-choice feeding assay, the total larval mass after 6 d was approximately 38.8% lower in larvae fed on overexpression plants and 51.0% higher in larvae fed on RNAi plants than in those fed on WT plants. Physiological analysis showed that RNAi plants accumulated significantly more MDA than WT and overexpression plants, whereas overexpression plants had higher chlorophyll a, chlorophyll b, and carotenoid contents than the other genotypes. Phytohormone analysis further showed that PhWRKY23-overexpressing plants accumulated higher levels of jasmonic acid, jasmonoyl-L-isoleucine, and salicylic acid, whereas abscisic acid showed no significant difference among genotypes. Yeast two-hybrid screening identified several candidate PhWRKY23-interacting proteins, and pairwise validation confirmed that PhWRKY23 interacted with PhDOX1 in yeast. These results indicate that PhWRKY23 positively contributes to herbivore resistance in P. hopeiensis and that this resistance phenotype is associated with changes in JA, JA-Ile, and SA accumulation and defense-related physiological traits.