The pinewood nematode (PWN), Bursaphelenchus xylophilus, is a causative agent of pine wilt disease (PWD). To date, although several molecular diagnostic methods have been developed, rapid on-site diagnostic tools for detecting PWN in pinewood are limited. In this study, a point of care diagnostic (POCD) method for detecting PWN in pinewood using recombinase polymerase amplification (RPA) assay was developed. This method comprises quick gDNA extraction buffer (DAP buffer) for the direct extraction of gDNA of PWN from pinewood and a battery-mounted portable optical isothermal device (POID) for the detection of PWD in the field. The RPA assay can distinguish between the PWN and its conspecies which exist in pinewood and can complete diagnostic procedures within 25 min in the field. Moreover, the RPA assay can detect PWN in old wood samples in both natural and stored conditions. The POCD-RPA assay to detect PWN will be useful for epidemiological investigations in the field as well as for quarantine processes in the wood trade.

Pine, an evergreen conifer, is widely distributed worldwide. It is economically, scientifically, and ecologically important. However, pine wilt disease (PWD) induced by the pine wood nematode (PWN) adversely affects pine trees. Many studies have been conducted on the PWN and its beetle vectors to prevent the spread of PWD. However, studies providing a comprehensive understanding of the pine tree transcriptome in response to PWN infection are lacking. Here, we performed temporal profiling of the pine tree transcriptome using PWD-infected red pine trees, Pinus densiflora, inoculated with the PWN by RNA sequencing. Our analysis revealed that defense-responsive genes involved in cell wall modification, jasmonic acid signaling, and phenylpropanoid-related processes were significantly enriched 2 weeks after PWD infection. Furthermore, some WRKY-type and MYB-type transcription factors were upregulated 2 weeks after PWD infection, suggesting that these transcription factors might be responsible for the genome-wide reprogramming of defense-responsive genes in the early PWD stage. Our comprehensive transcriptome analysis will assist in developing PWD-resistant pine trees and identifying genes to diagnose PWD at the early stage of infection, during which large-scale phenotypic changes are absent in PWD-infected pine trees.

Using a fumigation sterilizer (Dazomet) for nematode control, this study determined the effect of soil sterilization on the quantity and quality of the growth of Pinus densiflora Siebold & Zucc. and Larix kaempferi (Lamb.) Carrière seedlings in an open-field nursery at Jeongseon-gun, Kangwon-do, Republic of Korea. Most soil nematodes disappeared after fumigant sterilization. The organic matter content, cation exchange capacity, and Mg²⁺ content of the soil increased significantly, but the electrical conductivity (EC) decreased. The quantity of pine and larch seedlings was highly correlated with the soil nematode density. The soil properties changed due to soil fumigation, which affected the growth of both species. Pine seedling growth was affected by the soil chemical properties, such as available phosphorus (AP), Ca²⁺, Mg²⁺, and EC, and the growth of larch seedlings was affected by the EC and nematode density. The quality of the pine seedlings improved, whereas that of larch did not. This was a result of the difference in their tolerance to soil salinity. The seedling quality index (SQI) of the two species was related to the EC, but the pine seedlings were more affected by the EC, indicating that the control of soil nematodes by fumigant treatment significantly increased the quantity of seedlings produced and partly improved their quality. Soil management using a fumigant treatment was more efficient for producing pine seedlings than larch. Our results can be used to improve agricultural and forestry economics.

Pine wilt disease caused by pine wood nematodes (PWN) have been reported to give severe damage in many countries worldwide. Many measures of PWN management have been implemented, including using the nematophagous fungus Esteya vermicola as a biological control agent. However, understanding on the interaction between this fungus and pine trees is still limited. This study aimed to evaluate the effects of resinous compounds on spore germination and mycelial growth of E. vermicola in vitro. Research results showed that resinous compounds from pine trees significantly affect spore germination and mycelial growth of E. vermicola. In which, (+)α-pinene, (-)α-pinene, and (-)limonene completely inhibited spore germination and mycelial growth, with 100% of spore germination inhibition rate (SGIR) and mycelial growth inhibition rate (MGIR). The mixture of 14 resinous compounds also indicated potent inhibition of spore germination and mycelial growth of E. vermicola, with 100% and 91.8% SGIR and MGIR, respectively. These results suggest that trunk injection of spore suspension of E. vermicola to pine tree could be unsuccessful and ineffective for the control of pine wilt disease.

In South Korea Bursaphelenchus xylophilus the pine wood nematode was first reported in Pusan city, South Korea in 1988. Bursaphelenchus xylophilus causes pine wilt disease and is recognized as a ca...