In order to identify the specific antigens for pine wood nematode (PWN), 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 1,546 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 pecifically recognized PWN while two clones recognized 4 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. (This work was supported by National Institute of Forest Science)

In this study, we isolated and identified an aggregation-sex pheromone from Monochamus saltuarius, the major insectvector of the pine wood nematode in Korea. Adult male of M. saltuarius produces 2-undecyloxy-1-ethanol, which is knownto be an aggregation-sex pheromone in other Monochamus species. We performed field experiments to determine the attractivenessof the pheromone and other synergists. More M. saltuarius adult beetles were attracted to traps baited with the pheromonethan to unbaited traps. Ethanol and (-)-α-pinene interacted synergistically with the pheromone. Traps baited with pheromone+(-)-α-pinene+ethanol were more attractive to M. saltuarius adults than traps baited with pheromone, (-)-α-pinene, or ethanol alone.Ipsenol, ipsdienol, and limonene were also identified as synergists of the aggregation-sex pheromone for M. saltuarius adults.

Monochamus alternatus (M. alternatus) and Monochamus saltuarius (M. saltuarius) are major vectors for Bursaphelenchus xylophilus in South Korea. When an adult, they are easily distinguishable by several morphological classification. However, it is difficult to identification between M. alternatus and M. saltuarius when they are larvae as they have very similar morphological characters. Thus, they are not easily distinguishable without expertise about Cerambycidae taxonomy. Furthermore, during epidemiological investigation, sometimes, adults or larvae would not be founded in death pine trees. For these reasons, in this experiment, we are able to identified between M. alternatus and M. saltuarius by mitochondrial 12S rRNA gene primers that are specific to 12S rRNA gene fragment of M. alternatus using larvae tissue and frass. Moreover, we had examined whether vectors that were already escaped from dead pine tree have Bursaphelenchus xylophilus or not by multiplex PCR using larva frass that was remained in dead pine tree.

최근 5년 동안 식물검역 현장에서 검출된 뿌리썩이선충속 선충을 분류하여 Pratylenchus crenatus, P. fallax, P. kumamotoensis, P. panamaensis, P. penetrans로 동정하였다. 이 종들의 진단형질은 구순부의 주름 수, 두부의 형태, 구침의 길이, 수컷의 유무, 측대의 구조, 수정낭의 형태, 후부자궁낭의 길이, 미부의 형태 등을 포함한다. 이들의 사진과 측정치 및 형태적 특징을 기술하였다.

The genus Pratylenchus is considered as one of the most important plant-parasitic nematodes around the world because of its broad spectrum host ranges and worldwide distribution. 72 species of root-lesion nematodes have already been described from various parts of the globe. Among all of them, only 5 species are regulated as a target species in quarantine inspection. As the importation of target items inspecting nematodes are surely increasing every year, detection rate of nematodes in quarantine also increased. However, the rate of species identification has been decreased due to lack of related information. The species belonging to genus Pratylenchus are shown very low identification rate because we have specimens and information about only 10 domestic species and 5 regulated species. In this study, we identified the species in the genus Pratylenchus and obtained related information for application in the quarantine field. Species identification was performed with extracted nematodes from plants obtained in the recent quarantine inspection. Newly identified species were 5 species including Pratylenchus crenatus, P. fallax, P. kumamotoensis, P. panamaensis and P. penetrans. The photos, measured value and morphological characters of each species are provided.

The pinewood nematode (PWN, Bursaphelenchus xylophilus) is known to be a major pathogen of the pine wilt disease (PWD). However molecular pathology of B. xylophilus is not completely understood, the pathogenecity of PWD is related to cell wall-degrading enzymes such as endoglucanases, expansins and pectate lyases (PELs). Recently, we developed stage-specific expressed tag library of B. xylophilus and identified a novel PEL, Bx-PEL3. We cloned Bx-PEL3 gene with RT-PCR, which showed high similarity to previously reported Bx-PELs. Phylogenetic analysis revealed that PEL3 was much closer to PELs of B. xylophilus than any other PELs. PEL3 has a conserved intron site as found in Bx-PEL2 in the genomic DNA analysis. Quantitative real-time PCR analysis revealed that Bx-PEL1 and Bx-PEL2 were more predominantly expressed than the Bx-PEL3 in B. xylophilus. The difference of expression level among Bx-PELs according to growth condition suggests that each Bx-PEL plays different biochemical role in the pathogenesis of the PWD.

There are increasing interests in developing methods specifically detecting pathogenic Bursaphelenchus xylophilus. In order to develop a detecting method for B .xylophilus, at first we generated monoclonal antibodies (MAbs) specific to B. xylophilus, discriminating from other pine tree resident nematodes. Among 2304 hybridoma fusions screened. We finally selected a MAb clone, 9F10 and used for further study. To identify the antigenic target of MAb-9F10, we employed several biochemical methods such as SDS-PAGE, 2 dimensional electrophoresis, anion exchange chromatography, and immunoprecipitation to separate and isolate an antigenic target. Proteins from above methods were analyzed via nano-LC-ESI-Q-IT-MS. Peptides of GaLECtin were always detected from several proteomic analyses, suggesting that GaLECtin is the antigenic target of MAb-9F10.

콩 품종 Essex와 PI 437654간 교잡 후 F2 유래 F3 계통들을 재료로 하여 작성된 RAPD 유전자지도상에 cyst 선충 race 5에 대한 저항성 QTLs 분석을 실시한 바 결과를 요약하면 다음과 같다. 1. 회귀분석 결과 26개의 marker들(22 RAPD, 4 RFLP)에서 cyst 선충 race 5 저항성 반응에 대한 유의성이 인정되었다. 2. MAPMAKER /QTL 분석 결과2개의 저항성 QTL들이 탐색되었는데, 이 QTL들은 2개의 linkage groups(LGC-20와 Group 2)에 위치하였다. 3. 탐색된 2개의 QTL들 중 1개는 우성유전, ？고 나머지 하나는 열성유전양상을 나타내었다. 4. 콩 cyst 선충 race 5의 저항성에 대한 유의성이 인정되는 5개의 marker들간 상호작용을 알아보기 위한 다중회귀분석 결과 총 26개의 조합들 중 4개의 marker들(E023 , G101 , W03, pK418C)로 구성된 조합에서 가장 높은 표리적 변이의 값(35.2%)을 나타내었다.