Pine wilt disease caused by Bursaphelechus xylophilus has continued to expanding in Korea since its first occurrence identified from Busan in 1988. The direction of spread and density of two vector insects, Monochamus saltuarius and M. alternatus, are closely associated with the expansion and occurrence of pine wilt disease being observed in Korea. In 2016, a pheromone trap was installed to investigate the density of each vector. A correlation between the density of two insect vectors and the number of trees infected by pine wilt disease was further investigated in Gyeongnam, Gyeongbuk, Ulsan, and Jeju. To perform a correlation analysis, the number of infected trees was counted within a radius of 50m from which the pheromone traps were installed. In addition, factors that might have an effect on the density of vectors were investigated. This includes the altitude, temperature and forest type from the site where the pheromone traps were installed.

소나무재선충과 근연종인 Bursaphelenchus 속 2종간의 매개충 채내 침입력과 경쟁력을 비교 실험하였다. 공시충은 Bursaphelenchus xylophilus (이하 Bx), B. mucronatus (이하 Bm), B.thailandae (이하 Bt) 선충 3종과 솔수염하늘소 (Monochamus alternatus)와 북방수염하늘소 (M. saltuarius) 2종을 사용하였다. 매개충 번데기에 선충 3종을 10,000 마리씩 단독 또는 혼합접종을 하고, 우화 후 체내 선충 개체수를 조사한 결과, 솔수염하늘소에서는 Bx, Bm, Bt가 각각 2,283마리, 1,575마리, 3,083마리로써 Bt의 침입력이 가장 높게 나타났다. Bx+Bt 처리 시에는 Bx 24%, Bt 76%, Bx+Bm 처리 시, Bx 68%, Bm 32% 비율로 조사되어 Bt>Bx>Bm 순으로 침입력이 강한 것이 확인되었다. 북방수염하늘소 에서 Bx, Bm의 침입력을 비교한 결과 Bx는 2,120마리, Bm 1,730마리로 솔수염하늘소에서와 유사한 결과를 나타내었다. 이로써 선충 종간 침입력은 다양하며 이는 매개충 종과는 무관함을 알 수 있다.

Beetles of the genus Monochamus Dejean (Coleoptera: Cerambycidae) are the most important vectors of the pine woodnematode (PWN), Bursaphelenchus xylophilus (Steiner & Buhrer) (Nematoda: Aphelenchoididae). The most efficient wayto control pine wilt disease is to decrease the population levels of the vector Monochamus beetles. Specific and efficientnatural enemies would be an interesting and environmental-friendly option. From 2016 to 2017, we investigated on thelarval parasitoids associated with M. alternatus Hope and M. saltuarius Gebler using the sentinel logs (Pinus koraiensis)in South Korea. As results of this work, we report two larval ectoparasitoids, Spathius verustus Chao and Cyanopterusflavator (Fabricius) (Hymenoptera: Braconidae) from six survey sites. Each parasitoid was parasitized up to 58.2% and26.3% on 1st and 2nd instar larvae of Monochamus beetles, respectively. These parasitoids are likely to be highly efficientas bio-control agents. Morphological characters with illustration and some biological information of them are provided.

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.

This study was performed to investigate the escape of pine wood nematode (PWN), Bursaphelenchus xylophilus, from two vector species (Monochamus alternatus and M. saltuarius) through oviposition and feeding behavior. First, we checked number of PWNs escaped from M. alternatus emerged from three different cases of pine logs. In case A, healthy pine trees were cut into logs and left in pine forest infected with PWN. In case B, healthy pine trees were cut into logs, left in large screen cage, and let them oviposited by M. alternatus emerged from pine trees infested with PWN. In case C, pine trees which were harboring M. alternatus were cut into logs, and PWN was inoculated artificially. The M. alternatus adults emerged from the above three cases of pine logs were checked in the next year to know how many PWN they were harboring in their bodies. The percentages of M. alternatus harboring PWN (18.3 and 15.6%, respectively) and number of nematodes per vector (5,713.1±9,248.3 and 2,034.1±4,746.8 PWNs, respectively) in case A and B logs are similar to each other. However, the percentage and the number in case C (38.3% and 20,083.1±32,188.3 PWNs) were higher than those of case A and B. Among 52 M. alternatus adults harboring PWN from all the three cases, 20 adults (38.5%) were harboring more than 5,000 PWNs per beetle. And these 20 adults were harboring 97.9% of the total PWNs in 52 adults. Second, we checked the daily escape of PWNs from M. alternatus and M. saltuarius collected at pine forest infested with PWN. The PWN escaped from their vector body for 34.9±12.4 days for M. alternatus, and for 23.9±16.2 days for M. saltuarius, reaching at peak escape during the 2nd week of emergence of the two vector species. A 44.5 and 47.2% to the total PWNs escaped from vector body within 2 weeks of vector emergence for M. alternatus and M. saltuarius, respectively. The number of PWNs escaped from each vector was 3,570.6±5,189.2 and 1,556.2±1,710.3 for M. alternatus and M. saltuarius, respectively.

Trunk injection of nematicides is applied to prevent pine wilt disease(PWD). Although the trunk injection of nematicides is effective to PWD prevention, it is not effective to vector insects such as adults of M. alternatus and M. saltuarius. Thus, This study was investigated for the insecticidal activity and the effect of feeding deterrence by injecting to pine trees with acetamiprid. For screening effective nematicides, some insecticides were injected to pines on December 30th, 2008 and their insecticidal activities and feeding areas of M. alternatus and M. saltuarius were investigated in the insects breeding dishes (100× 40mm) where put on 1 to 2-year-old twig which was selected from injected pine trees in early June. As a result, insecticidal activity and feeding deterrence of acetamicprid were most effective. Feeding areas of M. alterantus adults, when tested with 1-year-old twig and 2-year-old twig of healthy pine trees, were 170.3±107.0 mm2 and 258.0±123.2 mm2 respectively. And Feeding areas of M. alterantus adults, when tested with 1-year-old twig and 2-year-old twig of pines injected at a dose of 0.3 ml per cm dbh of a pine tree with acetamiprid, were 57.7±72.3 mm2 and 65.7±51.2 mm2 respectively and their mortalities also were 0%, 28.6%. The feeding areas of M. alterantus adults tested with 1-year-old twig and 2-year-old twig of pine trees injected at a dose of 0.6 ml per cm dbh of a pine tree with acetamiprid, were not fed on it and the mortalities were 28.0%, 57.1% respectively. In case of the feeding areas of M. saltuarius adults, when tested with 1-year-old twig and 2-year-old twig of healthy pine trees, it was 112.±35.1 mm2 and 159.2±65.2 mm2 respectively. And the feeding areas of M. saltuarius adults, when tested with 1-year-old twig and 2-year-old twig of injected pine trees at a dose of 0.3 ml per cm dbh of a pine tree with acetamiprid were 0.61±0.0 mm2 and 8.1±12.4 mm2 and both mortalities were 23.1%. Although M. saltuarius adults, when tested with 1-year-old twig of a pine tree injected at a dose of 0.6 ml per cm dbh of a pine tree with acetamiprid, were not fed on it but M. saltuarius adults tested with 2-year-old twig of the same pine tree were 2.0±3.7 mm2 and their mortalities were 53.8%, 61.5% respectively. We investigated insecticidal activities, the effects of feeding deterrence of acetamiprid for vector insects where fed twigs of injected pines by stop up hole with a cork or not, and the trunk injection to pine trees at a dose of 0.5, 1 ㎖ per ㎝ dbh of a pine tree with acetamiprid on the early June. As a result, all of the vector insects were not fed and the feeding area was within 11 ㎟ and the mortality of M. alternatus was 70%.

Pine wood nematode, Bursaphelenchus xylophilus, is a causative organism to induce pine wilt disease in many varieties of pine trees. Until 2006, Monochamus alternatus had been known as the only insect vector of pine wood nematode in Korea which targeted on Pinus densiflora (Japanese red pine) and P. thunbergii (Japanese black pine). However, pine wilt disease was also reported from Korean pine tree (Pinus koraiensis) in 2006 and we found another insect vector, M. saltuarius, was involved to transmit pine wood nematode. Both Monochamus species were confirmed to transfer pine wood nematode to their hosts but, there is no detail information about other transmitted nematode. Especially Bursaphelenchus mucronatus is common species transmitted by Monochamus species which is morphologically closed to B. xylophilus. Moreover B. mucronatus have two genotypes; one is East Asian type and the other is European type. Both genotypes of B. mucronatus were found in Korea but, the host and vector information related to the genotypes of B. mucronatus was not clear. Monochamus saltuarius was collected from three different geographical locations and nematodes were extracted and identified. For the identification of the juveniles, nematode DNA was extracted and ITS-RFLP analysis was done by PCR and gel electrophoresis. The selected enzymes were Hinf I, Alu I, Msp I, Hae III, Rsa I. Most of Bursaphelenchus species carried by M. saltuarius, which collected from pine wilt disease-free area, was determined as European type of B. mucronatus. We will compare the nematode species and genotypes carried by M. alternatus and M. saltuarius. In addition the rate of nematode carrying insect and the average number of nematode per single insect will be counted and compared.

A survey was conducted in the northern conifer forests of Korea from February to May in 2009 for the Monochamus species vectored Bursaphelenchus species. Dead pine trees infested with bark beetle larvaes were collected from Pinus densiflora and P. koraiensis located in the following target areas: Pocheon, Yeongi, Jincheon, Chuncheon, and Bonghwa. A cerambycid beetle, M. saltuarius was only found from sampled log piles in target areas. Bursaphelenchus species carried by M. saltuarius was only isolated in Pocheon, Gyeonggi province. The nematodes recovered from 17 (71%) of 24 beetles from P. koraiensis and 12 (75%) of 16 beetles from P. densiflora. The average number of nematode per adult was 1,974 and 11,823 from P. koraiensis and P. densiflora, respectively. The nematode was also recovered from the inner bark of both Pinus species infested with M. saltuarius larvae. Bursaphelenchus species identification was performed by morphological characters and molecular analysis with ITS-RFLP patterns and sequences of ITS and D2D3 region. Both morphological and molecular characters match well with the original description of the European type of B. mucronatus. This is the first report of M. saltuarius species as a vector of the European type of B. mucronatus on both P. densiflora and P. koraiensis in Korea.

Monochamus alternatus and M. saltuarius were reported as the vectors of Bursaphelenchus xylophilus, pine wood nematode in Korea. According to Kwon et al. (2006), each of 2 species has occupied their own regional distribution : M. saltuarius in southern part including Jeju island and M. alternatus in mid-northern part of Korean peninsula. We measured the supercooling point (SCP) of 2 species (laboratory-reared populations) by each of developmental stages. The SCPs of 2nd, 3rd and 5th instar larvae of M. saltuarius were -7.68±0.19℃, -7.02±0.69℃, -4.93±1.34℃ each of stages. On the other hand, the SCPs of 3rd, 4th, 5th instar larvae and pupae of M. alternatus. were -4.46±1.12℃, -5.94±1.33℃, -7.83±1.44℃, -9.53±1.78℃ each of stages. The SCPs of M. saltuarius larvae generally was lower than that of M. alternatus. The pupae of M. alternatus and 2nd instar larvae of M. saltuarius had the lowest SCP among measured samples. On the other hand, the highest SCP were recorded in 2nd and 5th instar larvae, each. This result shows that regional distribution of 2 beetles may be associated with the adaptation capacity to low temperature represented by the SCP as well as the developmental temperature. However, beetles experimented were not collected from pine forest fields. In further study, we are planning experiments with field populations and all developmental stages.

As a major component of cuticle wax, cuticular hydrocarbon plays key roles as a chemical cue among inter- and intra-species in some beetles like as pheromone and alleolochemicals in addition to the physical function. In this study, cuticular hydrocarbon was analyzed and compared with three species of beetles adult, Monochamus alternatus, M. saltuarius belong to the Monochamus sp., and belong to the same class, Moechophyta diphysis, a typical vector of pine wilt disease. The composition of cuticular hydrocarbon of three species of beetles adult did not show the difference between male and female in intra-species, however, showed the difference in inter-species. They also differed from carbon numbers in inter species as 25-32 in M. saltuarius, 25-35 in M. alternatus, and 23-31 in M. diphysis. Major constituent of M. saltuarius was analyzed as n-C25, 2 or 4 MeC26, 9-C25:1, n-C27, 4-MeC28, n-C29, (9,x)-,and/or (11,x)-diMeC29; those of M. alternatus were n-C25, n-C27, 4-MeC28, n-C29, (9,x)-, and/or (11,x)-diMeC29, 9-C29:1, n-C29, (9,x)-,and/or (11,x)-diMeC29; and those of M. diphysis were n-C25, n-C26, n-C27, 3-MeC27,9-C29:1, 11-MeC29. The contents of n-alkanes were as follows: M. saltuarius ≒ M. alternatus > M. diphysis. The contents of monomethylalkanes were as follows: M. diphysis > M. saltuarius ≒ M. alternatus. The contents of dimethylalkanes were as follows: M. saltuarius ≒ M. alternatus > M. diphysis. And the content of olefine in female were analyzed as follows: M. saltuarius > M. alternatus ≒ M. diphysis, and the male's contents were similar in three kinds of beetles.