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안세희 ( Sye Hee Ahn ),전문장 ( Mun Jang Jeon ),엄영근 ( Young Geun Eom ),오세창 ( Sei Chang Oh ),이미림 ( Mi Rim Lee ) 한국목재공학회 2011 목재공학 Vol.39 No.1
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.
소나무재선충 생물학적 방제를 위한 Bacillus licheniformis MH48의 선발 및 특성 규명
정민해 ( Min Hae Jeong ),양서영 ( Seo Young Yang ),이용성 ( Yong Sung Lee ),안영상 ( Young Sang Ahn ),박윤석 ( Yun Serk Park ),한혜림 ( Hye Rim Han ),김길용 ( Kil Yong Kim ) 한국산림과학회 2015 한국산림과학회지 Vol.104 No.3
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.
Cryopreservation of Pine Wood Nematode, Bursaphelenchus xylophilus
박주용,신금철,김경희,이승규,이종규 한국산림과학회 2005 한국산림과학회지 Vol.94 No.5
; 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?5.3%. The survival ability of nematodes at juvenile stage was more efficient than that of nematodes at adult stage. Pre-incubation treatment at 4oC 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 28oC 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.
Kim, Eun-Shik,Lee, So-Hee,Kim, Joon-Bum,Kim, Chan-Soo,Yoon, Bong-Taek,Lee, Sung-Hoon,Lim, Wontaek,Kim, Hyojung,Choi, Junghwan,Han, Hyerim The Ecological Society of Korea 2015 Journal of Ecology and Environment Vol.38 No.1
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.
Lee Jae-Woo,Nam Il,Park Ji-Hong,Huh Min-Jung,박일권 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.3
In this study, we investigated the nematicidal activities of the ethanol extracts of 49 medicinal plants against the pine wood nematode, Bursaphelenchus xylophilus, and isolated a main nematicidal constituent, (Z)-ligustilide, from Angelica tenuissima Nakai root extract. Among the 49 plant extracts, only the A. tenuissima root extract showed the strong nematicidal activity against the pine wood nematode, with a 92.3% mortality rate at a concentration of 2 mg/mL. Based on bioassay-guided isolation and gas chromatography-mass spectrometry (GCMS) analysis, (Z)-ligustilide was identified as the active component of A. tenuissima root extract at 73.6% of the total content ratio. The LC 50 value of (Z)-ligustilide against the pine wood nematode was 0.24 mg/mL. Our results indicated that (Z)-ligustilide as well as A. tenuissima root extract can be potential candidates for novel trunk injection agents against the pine wood nematode.
김아영,Angelina F. Osabutey,윤경재,최보혜,이시혁,한혜림,고영호 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.1
In order to identify the specific antigens for pine wood nematode (PWN, Bursaphelenchus xylophilus [Steiner and Buhrer] Nickle), we confirmed that one of the genes commonly found in the transcriptome, proteome and secretory proteins of PWN belonged to the Aldose Reductase (AR) family protein. 36.5 kDa PWN-AR1 was expressed and purified using Baculovirus Expression System. Total 1546 hybridoma fusion library was generated and screened for specificity to PWN-AR1 by Enzyme Linked Immunosorbent assay (ELISA). Nine clones showed strong immunoreactivity to PWN-AR1 were limited-diluted. Total 864 limited-diluted clones were further screened using PWN-AR1 by ELISA and 34 monoclonal antibody (Mab) clones were selected. 34 Mab clones were further screened using PWN extracts and a standard PWN-infected pine tree extract by ELISA. Finally nine clones were selected and their immunoreactivities to 4 different nematodes were examined by ELISA. Seven clones specifically recognized PWN while two clones recognized four nematodes. Our data suggested that PWN-AR1 is a PWN secretory enzyme while PWN is invading pine trees, Thus, PWN-AR1-Mabs could be used to develop diagnosis tools for PWN and its infected pine trees.
Cryopreservation of Pine Wood Nematode, Bursaphelenchus xylophilus
Park, Ju-Yong,Shin, Keum-Chul,Kim, Kyung-Hee,Lee, Seung Kyu,Lee, Jong Kyu Korean Society of Forest Science 2005 한국산림과학회지 Vol.94 No.5
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.
Hyang-Mi Cheon,Hye-Soon Huh,Hee-Sun Rho,Kwang-Sik Choi 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.05
The ovarian development and oviposition response of two vector beetles of pine wood nematode, Monochamus alternatus and M. saltuarius, was investigated. The ovary structure of two beetles was investigated by means of light microscopy. The ovary of both beetles was composed of two ovaries that are connected by a common oviduct. Each of the two ovaries was consisted of twelve ovarioles, the functional units of female oogenesis. The ovary type was meroistic telotrophic with nurse cells and oocytes in the tropharium. Ovarian development of two vector beetles was completed at 12-14 days after maturation-feeding pine twigs. Aspect of accumulation of vitellogenins in the ovary of two vectors showed difference in developmental stages and major yolk proteins, differently from vitellogenin. To investigate oviposition performance of two vector beetles, we provided P. thunbergii and P. koraiensis bolts for egg laying and collected emerged adults from each pine bolts in the next year. M. saltuarius females made more oviposition wounds and entrance hole of larvae than M. alternatus on pine bolts. We also investigated whether two beetles can transmission of Bursaphelenchus xylophilus and the European type of B. mucronatus via oviposition to noninfected dead pines. 48-83% from newly emerged adults of two beetles were carried two species nematode. This results suggested that two species beetles can transmit nematodes through oviposition performance and transmitted nematodes successfully propagate in non-infected dead pines.
Suppression of pine wilt disease by an antibacterial agent, oxolinic acid
Kwon, Hyeok Ran,Choi, Gyung Ja,Choi, Yong Ho,Jang, Kyoung Soo,Sung, Nack-Do,Kang, Mun Seong,Moon, Yilseong,Lee, Seung Kyu,Kim, Jin-Cheol John Wiley Sons, Ltd. 2010 PEST MANAGEMENT SCIENCE Vol.66 No.6
<P>BACKGROUND: Pine wilt disease (PWD) is very complex and has been reported to be caused by pine wood nematode, Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle, and its accompanying bacteria. However, there is no report on the control of PWD by antibacterial agent. The present study was performed to investigate disease control efficacy of antibacterial agents against PWD.</P><P>RESULTS: Among six antibacterial antibiotics tested, oxolinic acid (OA) showed the strongest antibacterial activity against five bacteria isolated from three strains of pine wood nematode. In in vivo assay, it effectively suppressed the development of PWD in three-year-old seedlings of Pinus densiflora Sieb. & Zucc.; it showed 71% control when injected at 3 mg per seedling. A mixture of OA and the nematicidal agent abamectin (Ab) showed higher disease control efficacy against PWD than either OA or Ab alone. In addition, OA alone and a mixture of OA and Ab also controlled PWD in approximately 20-year-old pine trees under field conditions.</P><P>CONCLUSION: This is the first report on the suppression of PWD by OA. The result strongly indicates that PWD could be controlled by antibacterial antibiotic alone and a combination of antibacterial and nematicidal agents. Copyright © 2010 Society of Chemical Industry</P>