Enhanced accumulation of pinosylvin stilbenes and related gene expression in Pinus strobus after infection of pine wood nematode

Pine wood nematodes (PWNs; Bursaphelenchus xylophilus) infect pine trees and cause serious pine wilt disease. Eastern white pine (Pinus strobus) has resistance to PWN. However, the detailed defense mechanisms of P. strobus against PWN are not well known. When P. strobus Plants were infected with PWNs, the accumulation of stilbenoids, dihydropinosylvin monomethyl ether (DPME) and pinosylvin monomethyl ether (PME) was increased remarkably. Both DPME and PME had high nematicidal activity. The nematicidal activity of the two compounds was resulted in a developmental stage-dependent manner. Pinosylvin monomethyl ether was more toxic to adult PWNs than juveniles, whereas DPME was found more toxic to juvenile PWNs than the adults. The genes involved in PME and DPME biosynthesis such as phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase (4CL), pinosylvin synthase (STS) and pinosylvin O-methyltransferase (PMT) were isolated using de novo sequencing of the transcriptome in P. strobus. In addition, transcription factors (TFs; bHLH, MYB and WRKY) related to stilbene biosynthesis were isolated. qPCR analyses of the selected genes (PAL, 4CL, STS and PMT) including TFs (bHLH, MYB and WRKY) revealed that the expression level of the selected genes highly enhanced after PWN Infection. Our results suggest that pinosylvin-type stilbenoid biosynthesis is highly responsive to PWN Infection and plays an important role in PWN resistance of P. strobus trees.