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Pine wilt disease has caused great economic loss and become an ecological threat since it was introduced into East Asia. In China, Pinus massoniana Lamb. is highly susceptible. The pathogenic of this disease is linked to the invasion of P. massoniana by the pine wood nematode (PWN, Bursaphelenchus xylophilus), leading to various physiological activities. However, the migration pathway of PWN and the defense mechanisms of P. massoniana tissue structure following invasion remain unclear. This knowledge is vital for understanding the pathogenesis of pine wood nematode disease. To address this issue, we analyzed the tissue structure damage, horizontal and vertical migration pathways, and histochemical reactions of P. massoniana after PWN inoculation. The results are as follows: susceptible P. massoniana exhibited more tissue structure damage compared to highly resistant P. massoniana. PWN reproduced and migrated by feeding on and damaging cells. In susceptible P. massoniana, PWN propagated and migrated throughout the entire plant. Highly resistant P. massoniana displayed limited horizontal and vertical migration of PWN, making it challenging for PWNs to move from cambium to xylem. After P. massoniana was damaged by PWNs, a protein cross-linking phenomenon appeared rapidly, with highly resistant P. massoniana exhibiting less protein cross-linking than the susceptible variety. Lignin synthesis is a crucial factor in the tissue defense of P. massoniana. Protein crosslinking provides time for lignin synthesis and is an vital component of tissue defense. [ABSTRACT FROM AUTHOR]

Pinus massoniana Lamb. is a crucial timber and resin conifer in China, but its plantation industry is threatened by outbreaks of pine wilt disease (PWD) caused by Bursaphelenchus xylophilus (pinewood nematode; PWN). However, as of yet, there is no comprehensive analysis of NBS-LRR genes in P. massoniana involved in its defense against PWN. In this study, 507 NBS genes were identified in the transcriptome of resistant and susceptible P. masoniana inoculated with the PWN. The phylogenetic analysis and expression profiles of resistant and susceptible P. massoniana revealed that the up-regulated PmNBS-LRR97 gene was involved in conferring resistance to PWN. The results of real-time quantitative PCR (qRT-PCR) showed that PmNBS-LRR97 was significantly up-regulated after PWN infection, especially in the stems. Subcellular localization indicated that PmNBS-LRR97 located to the cell membrane. PmNBS-LRR97 significantly activated the expression of reactive oxygen species (ROS)-related genes in P. massoniana. In addition, the overexpression of PmNBS-LRR97 was capable of promoting the production of ROS, aiding in plant growth and development. In summary, PmNBS-LRR97 participates in the defense response to PWN and plays an active role in conferring resistance in P. massoniana. This finding provides new insight into the regulatory mechanism of the R gene in P. massoniana. [ABSTRACT FROM AUTHOR]