State key R & D Program
为了进一步阐明细菌在松材线虫病中的作用，本研究采用松材线虫（CK）和松材线虫与马尾松内生细菌GD2 的混合物（T）分别接种马尾松，并对松材线虫进行了转录组测序， Gene Ontology（GO）及 KEGG 富集分析，并利用 Real-time Quantitative PCR(RT-qPCR)验证目标基因表达情况。结果显示，与对照组CK相比，共筛选到143个差异表达基因(differentially expressed genes，DEGs)，其中有 63 个上调基因，80 个下调基因。Gene Ontology 分析显示，差异表达基因在免疫系统过程、细胞膜、代谢过程、转运活性等功能类富集。KEGG 通路在ECM-receptor 互作、肾素-血管紧张素、糖酵解、胰岛素信号等通路富集。这些GO 功能与KEGG 通路涉及机体的免疫调节。选取 8 个基因进行RT-Qpcr 验证，表达水平与测序结果一致，证明了测序结果的准确性。上述分析为进一步揭示细菌增强了松材线虫对寄主的适应性分子机制提供了参考。
In order to further clarify the role of bacteria in pine wilt disease (PWD), Pinus Massoniana was inoculated with Bursaphelenchus Xylophilus (CK), a mixture of Bursaphelenchus xylophilus and the endophytic bacterium GD2(T) respectively. Transcription sequencing, Gene Ontology (GO) and KEGG enrichment analysis were performed on the Bursaphelenchus Xylophilus, and Real-time Quantitative PCR (RT-qPCR) was used to verify the target Gene expression. The results showed that 143 differentially expressed genes (DEGs) were detected, among which 63 were up-regulated and 80 were down-regulated. The analysis of Gene Ontology showed that the differentially expressed genes were enriched in immune system process, cell membrane, metabolic process, transporter activity and other functions. KEGG pathway was enriched in ECM-receptor interaction, Renin?angiotensin system, Insulin signaling pathway and other pathways. These GO functions and KEGG pathways were involved in immune regulation and reproductive development of the body. Eight genes were randomly selected for RT-QPCR verification, and the expression level was consistent with the sequencing results, which proved the accuracy of the sequencing results. The above analysis provides valuable reference for further revealing the molecular mechanism by which bacteria enhance the adaptability of pine wood nematode to its host.