• Mycobiology
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• v.35 no.3
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• pp.159-161
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• 2007

Pine tree death caused by pine wood nematode(PWN) involves phoretic relationships between PWN and its vector Japanese pine sawyer beetle(JPS). In an effort to understand the diversity of fungi involved in PWN life cycle, a total of 176 fungal isolates were collected from PWNs, adults and larvae of JPS, PWN-diseased Japanese black pine that was cut down in 2005 at Jinju, Korea. Based on microscopic observation and colony morphology, and sequence analysis of the ITS rDNA, the fungal isolates were identified at the level of genus. Three genera including Mucor, Ophiostoma, and Penicillium were identified from PWN. Two genera of Ophiostoma and Penicillium were discovered from JPS larvae. Frpm JPS adult beetles, nine genera of Aspergillus, Gibberalla, Hypocrea, Irpex, Leptosphaeria, Ophiostoma, Penicillium, and Plectosphaerella and unknown basidio-mycetes were found. Ten genera from PWN-infected weed were confirmed as Bionectria, Botrytis, Camarops, Fusarium, Hypocrea, Nectria, Mucor, Ophiostoma, Penicillium, and Trichoderma. Penicillium and Ophiostoma were commonly distributed on PWN and its vector and host. This is first report of the fungi associated with PWN and its vector and host in Korea.

• Journal of the Korean Wood Science and Technology
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• v.39 no.1
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• pp.15-20
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• 2011

The pine wilt disease is one of the most serious forest diseases that kill the pine trees, and the study on the invasion and movement of the pine wood nematode within the tree is very important for understanding the inhabitation of pine wood nematode. In this relation, the microscopic observation was carried out to study the place of inhabitation and movement of pine wood nematode within the infested wood. In result, the rays were mainly infested by pine wood nematode and showed dark discoloration due to their necrosis in cross, radial and tangential surface. Also, the intensive damage was found in the resin canals. On the other hand, some traumatic resin canals in tangential band were identified in the sapwood near the cambium. In the ray, the pine wood nematode occurred more commonly in the ray parenchyma cell and fusiform ray with horizontal resin canal than in the ray tracheid and uniseriate ray without horizontal resin canal, respectively. The pine wood nematode was thought to move from ray to tracheid through the large natural opening, window-like pit, in the cross-field, neither through the small natural opening, bordered pit, in the tracheid nor through the tracheid wall by creating a bore hole.

• Journal of Forest and Environmental Science
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• v.33 no.2
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• pp.105-112
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• 2017

The distribution and form of pine wood nematode (PWN) were investigated in Jinju-si and Sacheon-si areas, where infested with pine wilt disease (PWD). The average PWN population per g of the tubulation part in the wilted Japanese balck pine were 381.2 and 341 in Jinju-si and Sacheon-si, respectively. The PWN population per g of affected Japanese black pine with wilt rate below 60% were 556 and 518 in Jinju-si and Sacheon-si, respectively. The ratios of PWN and other plant-parasitic nematode in the wilted Japanse black pines were 48% vs. 52% in Jinju-si and 53% vs. 47% in Sacheon-si after exposure to PWD for 1 year. The survivorship of pine wood nematode in the sampled trees after 1 year was 0-10% in the Jinju-si area and 5-20% detected in the Sacheon-si area. In the forest areas affected by PWN, the highest survival rates were 37.2% and 39.8% at 25 cm diameter at breast height (DBH) in Jinju-si and Sacehon-si, respectively, while the highest wilt rates were 30.5% and 28.3% at 30 cm DBH in Jinju-si and Sacehon-si, respectively.

• Journal of Korean Society of Forest Science
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• v.94 no.5 s.162
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• pp.334-337
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• 2005

Pine wood nematode isolates from Haman, Gyoungsangnam-do were cryopreserved with three different cryoprotectants (glycerol, ethylene glycol and dimethylsulfoxide), Nematode Growth medium (NGM) and M 9 buffer solution, respectively, and then survival rate, reproduction ability, and pathogenicity of stored nematodes were compared. Survival rates of juvenile was excellent in 15% glycerol solution as $72{\pm}5.3%$. The survival ability of nematodes at juvenile stage was more efficient than that of nematodes at adult stage. Pre-incubation treatment at $4^{\circ}C$ before storing in liquid nitrogen didn't affect the survival ability of pine wood nematode. When the reproduction ability of nematodes was confirmed on Botrytis cinerea agar plates at $28^{\circ}C$ incubator, there was no difference between cryopreserved and non-cryopreserved. Pathogenic ability of cryopreserved nematodes was also evaluated and confirmed by artificially inoculating them on 2-year old red pine seedlings.

• Journal of Korean Society of Forest Science
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• v.104 no.3
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• pp.512-518
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• 2015

Pine wilt disease (PWD) caused by pine wood nematode, Bursaphelenchus xylophilus, has become the most serious threat to pine trees in Korea. This study was subjected to investigate effective biological control agent against PWD. To select nematocidal bacteria against PWD, Bacillus licheniformis MH48 was selected among five bacteria due to its high nematocidal potential. B. licheniformis MH48 was tested for cell growth and protease activity to evaluate its nematicidal potential. In the B. licheniformis MH48, cell numbers were highest three days after incubation, while protease activity was highest after seven days. In the effect of different concentrations of B. licheniformis MH48 culture broth against B. xylophilus, 20% concentration of culture broth showed approximately 80% of pine wood nematode mortality compared to the control. Especially, pine wood nematode's cuticle layers were degraded two days after treatment of B. licheniformis MH48 culture broth. The present study suggests that B. licheniformis MH48 can be one of the potential biocontrol candidates against pine wood nematode due to its ability to produce protease.

• Korean journal of applied entomology
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• v.29 no.1
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• pp.14-19
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• 1990

Nematodes and insects associated with dead trees were surveyed on the 238 dead trees belonging to 19 tree species in Pusan, Gyeongnam, Gyeongbuk, Chunnam, and Chonbuk provinces from April to September of 1989. Pine wood nematode, Bursaphelenchus xylophilus was found only in Pusan but B. mucronatus was collected at Chinju and Chinhae. The 13 nematode species in 9 genera were identified. Out of them, Diplogasteroides dimidius, Rhabdontolaimus adephagus, R. janae, Mikoletzkya diluta, M. ruminis, M. langcaudaa, Parasitorhabditis hylurgi, Panagrolaimus concolor, Panagrodontus dentatus, Prothalonema intermedium, and marcrolaimus canadensis were recorded for the first time in Korea. Insects collected from dead trees were 5 orders, 9 families, 25 genera, and 27 species. of them, the Coleoptera were the most collected insects by the 3 families, 19 genera and 22 species. The Scolytidae were 12 species in 10 genera. Hypothenemus eruditus was firstly collected from Campylotropis macrocarpa, Lespedeza maximowizi, Forsythia ovata, Meliosma oldhami, Securinega suffruticosa, Broussonetia kazinoki, and Cornus walteri. The maximum number of pine wood nematode was separated from the abdomen of Monchamus alternatus, the pine woodnematode vector. The maximum number of nematodes per an adult of M. alternatus was 127,535, minimum 2,616, and average 42,817.

• Korean journal of applied entomology
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• v.57 no.1
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• pp.7-13
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• 2018

Currently, there is no available tool that rapidly diagnoses pine wood nematode (PWN)-infected pine trees in the field. In this study, we synthesized and purified PWN Galectin, which might be an antigen specific to PWN, using the Baculovirus expression system. We used PWN Galectin as an antigen for generating 1,464 fusion hybridoma cell lines secreting monoclonal antibodies (Mabs). Among them, we selected 62 fusion hybridoma cell lines showing high reactivity to PWN Galectin. We further selected 12 fusion hybridoma cell lines showing high reactivity to the standard PWN-infected pine tree phosphate buffered saline (PBS) extract. Additionally, two fusion hybridoma cell lines showing no or extremely low reactivity were used as controls. The selected fusion hybridoma cell lines were subjected to limiting dilutions for selecting and establishing Mab-secreting cell lines showing higher reactivity to the standard PWN-infected pine tree extract than to the standard normal pine tree PBS extract. Moreover, the selected fusion hybridoma cell lines were further selected based on their higher reactivity to PWN protein extracts than to three non-pathogenic nematode protein extracts. The Mab-secreting cell lines established in this study could be used to develop rapid diagnostic tools that can be used in the field or in laboratories for detecting PWN-infected pine trees or PWN.

• Journal of the Korean Wood Science and Technology
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• v.37 no.3
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• pp.200-207
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• 2009

As an effort to utilize the pine woods damaged by pine wilt disease, this study analyzed the density of pine wood nematode and compressive strength of damaged trees treated through soaking and fumigating. The density of nematode in Pinus densiflora and Pinus thunbergii woods soaked in wood vinegar and nematicide, and in seawater reduced greatly in 21 days and in 30 days, respectively. When reextracted nematode was injected artificially into healthy trees, infection was not observed after the lapse of six months, and the mean density of Pinus densiflora and Pinus thunbergii was $0.47g/cm^3$ and $0.54g/cm^3$, respectively, and their compressive strengths were $304kgf/cm^2$ and $363kgf/cm^2$, respectively. As to change in the density of pine wood nematode after fumigating, pine wood nematode was detected until the third month in both species, and those detected afterwards were found to be similar nematodes. After 24 months, the mean densities of Pinus densiflora and Pinus thunbergii were $0.54g/cm^3$ and $0.52g/cm^3$, respectively, and their compressive strengths were $353kgf/cm^2$ and $370kgf/cm^2$, respectively.

• The Korean Journal of Pesticide Science
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• v.20 no.1
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• pp.56-65
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• 2016

Pine wood nematode, Bursaphlenchus xylophilus is a fatal damaging pest, infecting pinewood trees and is one of the serious forest pest in Korea. Trunk injection is one of the most effective method to protect the live pine trees from nematode infection. The aim of this research was to reveal the efficacy of the nematicidal materials from 46 Kyrgyzstan, 88 Vietnam plant materials and 21 active materials (${\alpha}$-terpinene, ${\alpha}$-pinene, ${\beta}$-thujaplicin, cinnamaldehyde, eugenol, emodin, geraniol, limonone, methyl palmitate, matrine, myrcene, methyl gallate, nicotine, quassin, resveratrol, rotenone, thymol, thujaplicin, terthiophene, tuberstemonine, ${\gamma}$-terpinene) isolated from plants for using as trunk injection agents. Eclipta prostrata extract was highly effective against pine wood nematode in plant extracts and terthiophene has the best nematicidal activity from active materials isolated from plants. Foliar spray of terthiophene on Japanese black pine (Pinus thunbergii) tree reduced proliferation of inoculated pine wood nematode at 22 days after treatment, however no efficacy was found over 22 days. Eclipta prostrata extract and terthiophene can be used as nematicide for further nematicidal efficacy test against any other plant parasitic nematodes and trunk injection efficacy against pine wood nematode.

• The Plant Pathology Journal
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• v.24 no.2
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• pp.159-163
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• 2008

Accurate identification of pine wood nematode, Bursaphelenchus xylophilus is a prerequisite to diagnose the pine wilt disease. However, a fungivorous nematode, B. mucronatus is highly similar to B. xylophilus and it is difficult to differentiate these two species by morphological features. A molecular diagnosis method, ITSRFLP was applied for the identification of B. xylophilus and B. mucronatus from Korea. Genomic DNA was extracted from a single individual nematode and ITS DNA was amplified by PCR. The size of PCR product was approximately 900bp and the sequence data were obtained after cloning. Amplified ITS was digested by 5 different restriction enzymes (Rsa I, Hae III, Msp I, Hinf I, and Alu I) and provided a discriminatory profile for B. xylophilus and B. mucronatus. Besides, B. mucro- natus was determined to have 2 different genotypes, East Asian type and European type also clearly separated by Rsa I and Hae III digestion. European type of B. mucronatus is recently collected from Pinus koraiensis and has not been reported before. ITS sequnce data were analyzed by Restriction Mapper program and the result supported ITS-RFLP pattern. These data indicated that PCRRFLP method is an accurate and simple way for identification of Bursaphelenchus species.