Pine trees are ecologically important in Korea. They are seriously imperiled by Pine wilt disease (PWD), by pine wood nematode (PWN, Bursaphelenchus xylophilus). Here, we isolated and characterized bacterial endophytes (BEs) from pine trees in Korea for biological control of PWN using BE metabolites. Using culture-dependent approach BE isolates were extracted from three tissues (needles, stems, and roots) of four pine species across 18 sampling sites in Korea. Bacterial isolates were characterized into 389 distinct isolates based on 16S rDNA sequencing. Ethyl acetate crude extracts (CEs) of bacterial liquid cultures were prepared using ethyl acetate and screened for nematicidal activity against PWN. BEs (1,622 isolates) were isolated; their taxonomic binning resulted in 215 operational taxonomic units (OTUs). Analysis of species richness and Shannon’s diversity of the three tissues revealed that BEs colonized the needles more than the stem and root tissues. Furthermore, based on nematicidal activity screening of 389 isolates, 44 BEs were identified, with two isolates exhibiting a significant inhibitory activity against PWN. Taken together, these data revealed numerous nematicidal BEs in pine trees, providing new insights that can serve as an effective and promising alternative approach to combat PWD.

Pine tree, a dominant species in the flora of Korea is the most beloved tree in Korea. However, recent outbreak of pine wilt disease caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus threatens the vegetation of Pinus species seriously. Furthermore, Pinus densiflora and P. thunbergii, the predominant species in Korea forest are highly susceptible to PWN. Therefore, there has been an urgent demand for the development of a new nematocidal compounds to control PWN, which spurred the national research for the development of new compounds. To find novel sources for nematocidal agents, we used various plant resources for the development. In addition to the foreign plant extracts themselves, we also used the extracts of endophytes composed of over 300 endophytic fungi and 1,000 endophytic bacteria from plants which was reported to contain nematocidal activity. Several extracts of endophytes and plant extracts contained strong nematocidal activity, and the resources are analyzed to identify the active nematocidal compounds. These integrated approach of finding effective nematocidal compounds from plants could be a novel way to elucidate the sources for brand-new nematocidal agents.