Transcription Factor and Protein Regulatory Network of in Response to Pine Wilt Nematode Infection.

Pine wilt disease, caused by , is a highly destructive and contagious forest affliction. Often termed the "cancer" of pine trees, it severely impacts the growth of Masson pine (). Previous studies have demonstrated that ectopic expression of the gene from in notably enhances resistance to pine wilt nematode infection. To further elucidate the transcriptional regulation and protein interactions of the in in response to pine wilt nematode infection, we cloned a 1984 bp promoter fragment of the gene, a transient expression vector was constructed by fusing this promoter with the reporter gene, which successfully activated the expression. DNA pull-down assays identified PmMYB8 as a trans-acting factor regulating gene expression. Subsequently, we found that the PmACRE1 protein interacts with several proteins, including the ATP synthase CF1 α subunit, ATP synthase CF1 β subunit, extracellular calcium-sensing receptor (PmCAS), caffeoyl-CoA 3-O-methyltransferase (PmCCoAOMT), glutathione peroxidase, NAD+-dependent glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase 1, cinnamyl alcohol dehydrogenase, auxin response factor 16, and dehydrin 1 protein. Bimolecular fluorescence complementation (BiFC) assays confirmed the interactions between PmACRE1 and PmCCoAOMT, as well as PmCAS proteins in vitro. These findings provide preliminary insights into the regulatory role of PmACRE1 in 's defense against pine wilt nematode infection.