• 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.

• 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.

• The Plant Pathology Journal
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• v.23 no.2
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• pp.70-75
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• 2007

We conducted an inoculation test using nine open-pollinated families of pine trees to evaluate their susceptibility and mortality in different densities of pine wood nematode. Three-year-old nine open-pollinated pine families were inoculated with Bursaphelenchus xylophilus at levels of 3,000, 5,000, and 7,000 nematodes/seedling in greenhouse. There were no distinct patterns in latent period among three densities of B. xylophilus in all families. Most families showed the first disease symptoms of needle discoloration within 12-15 days after inoculation. However, open-pollinated progenies of Pinus densiflora showed the longest latent period because none of one-year-old needles were wilted until 14 days after inoculation with 5,000 and 7,000 nematodes. One-year-old needles were wilted earlier than current needles in all tested families with all densities of B. xylophilus. Current needles were not wilted until 14 days after inoculation in all seedlings. The mortality of all seedlings rapidly increased from 35 days to 49 days after inoculation, and all died within 80 days except two seedlings. A 3,000 nematodes/100 ${\mu}L$ with sterilized distilled water are enough to screen 3-year-old pine seedlings for resistance to B. xylophilus.

• 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.97 no.6
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• pp.570-575
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• 2008

This study was carried out to provide basic information on physiological changes of 4-year-old seedlings of Pinus thunbergii inoculated with pine wood nematodes by analyzing changes of symptom development, stem and needle water content and nematode populations in stem. Twenty days after inoculation, needles were discolored as an external symptom, and xylem drying and pith browning occurred at the above and below of the inoculation sites as an internal symptom. However, xylem drying began to occur 10 days after inoculation, which was determined by the difference in drying status of xylem and cortex between control and inoculated seedlings. Although population of pine wood nematode increased from 5 to 10 days after inoculation, it has increased dramatically from 10 to 20 days after inoculation when both internal and external symptoms appeared. As the time passed by after inoculation with pine wood nematodes, water content of stem and relative water content in current needles and branch gradually decreased. As the number of nematodes increases, water content of stem and relative water content of current needles and branches decreased significantly. There was a positive relationship between the number of nematodes and xylem drying and/or disease development, but the number of nematodes rapidly decreased as seedlings become severely diseased.

• The Plant Pathology Journal
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• v.29 no.3
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• pp.344-349
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• 2013

Intraspecific variation in morphometry of pine wood nematodes Bursaphelenchus xylophilus and B. mucronatus in relation to geographical locations in South Korea was investigated using morphometric characters (body length, a, b and c ratio, stylet length, and spicule length for a male nematode and V (%) value for a female nematode). B. xylophilus was isolated from Pinus thunbergii in Jinju (1998), Ulsan (2000), Yangsan (2000), Mokpo (2001) and Jeju (2004), and from P. densiflora in Gumi (2001). B. mucronatus was isolated from P. thunbergii in Jinju (1991) and from P. densiflora in Milyang (2001). The body length of male and female B. xylophilus had the highest coefficient of variability and showed significant differences among geographical locations. The V (%) value for female B. xylophilus showed the lowest coefficient of variability, changing little with geographical area and host plant. All morphometric characters in B. mucronatus except for stylet length and female body length showed no significant differences between locations or hosts, suggesting they may not be affected by geographical area or host plant.

• Journal of Ecology and Environment
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• v.38 no.1
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• pp.85-93
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• 2015

In 2013, the epidemics of pine wilt disease caused by the pine wood nematodes (Bursaphelenchus xylophilus) resulted in damages to the forests of black pine (Pinus thunbergii Parl.) trees in Jeju Island, Korea. Among the affected trees, an old black pine tree at Cave Temple on Mt. Sanbangsan was included and died due to the prevalence of pine wilt disease. The tree was on Mt. Sanbangsan, which was designated as a National Scenic Place with the Number 77 and was believed to be more than 400 years old in age. By examining the disc of the tree stem obtained from the height of 2 m, we counted the tree rings from 4 different directions and cross-dated the readings by comparing the records of drought simulated from the BROOK Model. Our analysis indicates that the tree seems to have grown since late 1860s. Contrary to the belief of the general public, we can conclude that the age of the tree was estimated to be at maximum 150 years, which means that it was not the same old tree as was shown in the painting of the Tam-Ra-Sun-Ryeok-Do (an old painting book for the Inspection Tour of Jeju Island) published in 1702. Discussion was extended to the life history of the tree in growth and leaning and the measures to protect the tree species from the damages of the pine wilt disease caused by pine wood nematodes.

• The Plant Pathology Journal
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• v.39 no.1
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• pp.75-87
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• 2023

The pine wood nematode (PWN), Bursaphelenchus xylophilus is a well-known devastating pathogen of economic importance in the Republic of Korea and other countries. In the Republic of Korea, trunk injection of nematicides is the preferred method of control. In this study, the efficacy of 16 locally produced formulations of emamectin benzoate against the PWN are compared through determining their sublethal toxicities and reproduction inhibition potentials. Nematodes were treated with varying concentrations of the tested chemicals in multi-well culture plates, and rates of paralysis and mortality were determined after 24 h. Reproduction inhibition potential was tested by inoculating pre-treated nematodes onto Botrytis cinerea, and in pine twig cuttings. Despite the uniformity in the concentration of the active ingredient, efficacy was contrastingly different among formulations. The formulations evidently conformed to three distinct groups based on similarities in sublethal activity (group 1: LC₉₅ of 0.00768-0.01443 mg/ ml; group 2: LC₉₅ of 0.03202-0.07236 mg/ml, and group 3: LC₉₅ of as high as 0.30643-0.40811 mg/ml). Nematode paralysis generally occurred at the application dose of 0.0134-0.1075 ㎍/ml, and there were significant differences in nematode paralysis rates among the products. Nematode reproduction was only evident at lower doses both on B. cinerea and pine twigs, albeit the variations among formulations. Group 1 formulations significantly reduced nematode reproduction even at a lower dose of 0.001075 ㎍/ml. The variations in efficacy might be attributed to differences in inert ingredients. Therefore, there is need to analyze the potential antagonistic effects of the large number of additives used in formulations.

• The Plant Pathology Journal
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• v.29 no.1
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• pp.116-122
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• 2013

To evaluate the mechanisms of pathogenicity and nonpathogenicity of Bursaphelenchus xylophilus and B. mucronatus isolated in South Korea, we used 4-year-old P. thunbergii seedlings and 20-cm long one-year-old stem cuttings of 5-year-old Pinus thunbergii, and studied distributions and multiplications of pine wood nematodes after inoculation. The distributions of B. xylophilus in the 20-cm pine stem cuttings were not significantly different from that of B. mucronatus. Conversely, the proliferation rate of B. xylophilus on mycelial mats of Botrytis cinerea was significantly different from that of B. mucronatus. The study using 4-year-old P. thunbergii seedlings also showed that B. mucronatus can migrate to distal portions of the pine seedlings the same as B. xylophilus, but the populations of B. xylophilus remaining in the pine seedlings were relatively larger than those of B. mucronatus. Therefore, we concluded that the pathogenicity of B. xylophilus could be strongly influenced by its ability to multiply.

• 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.