In this study, in order to develop a method to efficiently inject essential nutrients necessary for plant growth into wood chips, which are simply used as soil covering materials in the agriculture, landscaping and horticultural industries, the atmospheric pressure dipping method and the vacuum pressure impregnating method are used to improve the plant nutrients injectability and impregnation amount were comparatively analyzed. Nutrient ingredients and 8 major heavy metal contents of wood chips injected with nutrients were analyzed, and soil covering effects were examined by covering wood chips injected with nutrients on soil. Comparing the dipping method and the vacuum pressure impregnation method, it took about 48 hours or more to inject 1,500 g or more of the nutrient aqueous solution into 1 kg of wood chips in the dipping method, but the vacuum pressure impregnation method could be impregnated in about 5 minutes. Components of the impregnated nutrients were detected in proportion to the diluted concentration. As a result of covering the wood chips developed in this study on soil, they showed weakly acidic pH, and the heat insulation and moisturizing effects during the winter season were evaluated to be superior to those of uncovered soil. In the future, wood chips impregnated with nutrients are expected to contribute to the more efficient use of waste wood resources and the long-term supply of nutrients essential for plant growth, reducing excessive use of chemical fertilizers and reducing costs.

There are many other detection methods for Bursaphelenchus xylophilus. However, Baermann funnel method, PCR-based methods, which are laborious and time consuming processes that are unsuitable to rapid diagnose the Pine Wilt Disease (PWD) on the field. For these reasons, the aim of our experiment is not only to apply field diagnostic for pine wood nematode (PWN) but also to reduce total time for detection PWN in the pine trees by loop-mediated isothermal amplification (LAMP) with phosphate (Pi)-induced coloration reaction which could be yes or no answer for detection of PWN has not required UV detector but it just could be discriminated by naked eyes within 30 min. Genomic DNA was directly extracted from pine wood chips by Wood Chips Direct Lysis procedure which can be used for LAMP only after simple dilution. Our results suggest that LAMP-Pi detection method, simple and rapid method for detection of PWN, could be applied to the field diagnostic for PWD.

It is difficult to identification between Bursaphelenchus spp. and Pine Wood Nematode (PWN) by morphological characteristics without expertise about nematode taxonomy. Furthermore, Baermann funnel method, which is nematode extraction method from wood chips or soil, requires at least 24 hours to extract nematode that is unsuitable to rapid diagnose the Pine Wilt Disease (PWD). For these reasons, the aim of this experiment is not only to improve accuracy of a PCR based method but also to reduce total experiment time for detection Bursaphelenchus spp. and PWN in the wood chips of PWD infected pine tree. In this experiment, we had been employed two PCR primer sets, which were originated from PWN specific Internal Transcribed Spacer (ITS) sequence region and Bursaphenchus spp. universal mitochondrial Cytocrome Oxidase subunit I (mtCOI) sequence region in order to discrimination between Bursaphelenchus spp. and PWN at the same PCR reaction. This experimental procedure was able to reduce experiment time and cost as well as to improve accuracy of detection than previous PCR based detecting method by not using Baermann funnel method and commercial genomic DNA extraction kit but using direct pine wood chips lysis method.