A xylan-decomposing Gram-positive bacterium, Cellulosimicrobium cellulans DY-8, was isolated from the gut of a wood-feeding longicorn beetle, Moechotypa diphysis. To amplify a partial fragment of the GH 10 β-1,4- xylanase (XylC) gene of strain DY-8, two degenerated oligonucleotide primers were designed based on strictly conserved regions (WDVVNE and ITELLDV) in the GH family 10 xylanolytic enzymes. The full gene (1488-bp) of XylC, which was predicted to encode a protein consisting of 495 amino acids with a molecular mass of 52.0 kDa and a calculated pI of 6.49, was cloned by repeated DNA walking and nested PCR protocols. The results of a protein blast survey showed that XylC was a β -1,4-xylanase comprised of an N-terminal catalytic GH10 domain (from Ser48 to Leu338) and a C-terminal RICIN domain (from Tyr359 to Leu492). This overall structure of XylC was 57% identical to that of Actinoplanes sp. SE50/110 β -1,4-xylanase (Accession number: YP_006265966), which has not yet been biochemically characterized.
Cuticular hydrocarbons (CHCs) of the pine sawyer (Monochamus saltuarius), Japanese pine sawyer (M. alternatus) and oak longicorn beetle (Moechotypa diphysis) were analyzed by GC, GC-MS and compared. Monochamus beetles are typical vectors of pine wilt disease but Moechotypa diphysis, which belongs to the same family, is not. They possess different CHCs in carbon number: 23-25 in M. saltuarius, 25-32 in M. alternatus, and 23-29 in M. diphysis. In comparison to inter-species, these three species of adult beetles have different numbers and chains of constituents of CHCs. In comparison between male and female in intra-species, the quantities of CHCs show the difference but constituents are not. Major constituent of M. saltuarius were analyzed as n-pentacosane > n-nonacosane > n-heptacosane, those of M. alternatus were n-nonacosene > n-pentacosane > n-nonacosane, and those of M. diphysis were n-heptacosane > 13-methylheptacosane > 3-methylheptacosane. From the body surface, most saturated carbohydrates of 3 species beetles are composed of n-alkane (40.2 - 65.7%) and followed by olefines > monomethylalkanes that one or two double bonds in M. saltuarius and M. alternatus. Otherwise, M. diphysis have the difference in order of monomethylalkanes > olefins.
본 연구는 표고원목재배시 골목을 가해하는 털두꺼비하늘소(Moechotypa diphysis) 성충에 대해서 25종 monoterpenoid의 훈증독성, 접촉독성 및 산란기피효과를 조사하였다. 훈증독성은 20(equation omitted)/954 ml (공기)의 농도에서 1, 8-cineole, fenchone, pulegone, (equation omitted)-terpinene이, 10(equation omitted)/954 ml (공기)의 농도에서는 pulegone과 (equation omitted)-terpinene이, 그리고 5(equation omitted)/954 m1 (공기)의 농도에서는 pulegone만이 100%의 살충률을 나타내었다. 접촉독성은 pulegone만이 70%의 살충률을 보인 것 외에 대부분의 monoterpenoid에서 활성이 낮거나 없었다. 후각계를 이용한 기피반응은 1 (equation omitted)에서 bornylacetate, carvacrol, 1, 8-cineole, menthol은 기피반응을 보였고, 반면 citronellol은 유인반응을 보였다. 실내에서 수행한 산란기피효과는 25개의 monoterpenoid중 1,000ppm의 농도에서 carveol, geraniol, perillyl alcohol이 각각 82.1%, 78.3%, 87.5%의 효과를 보였다. 이 3 화합물을 가지고 야외포장 적용시험을 수행한 결과 10,000ppm과 1,000ppm의 농도에서 3일째까지 geraniol이 가장 효과가 좋았으나 잔효성은 없는 것으로 나타났다.