The pinewood nematode (PWN, Bursaphelenchus xylophilus) is known as a virulent factor of the pine wilt disease, transmitted to pinewoods by the pine sawyer beetle, Monochamus alternatus. It is very hard to discriminate B. xylophilus from B. mucronatus because these Bursaphelenchus species are genetically and biochemically very close. Therefore, it has been necessary to detect PWN-infected trees for the prevention of pine wilt disease transmission in a short time. We developed polyclonal antibodies against B. xylophilus in BalbC mice and primarily screened with ELISA. Positive clones releasing polyclonal antisera revealed B. xylophilus-specific immuno-reactivity, which were at least two times higher than that of B. mucronatus. Two clones, D9-F10 and 1F3, were finally selected and exhibited specific immuno-reactivity for B. xylophilus, not for B. mucronatus in Western blot analysis. D9-F10 clone was reactive with a 43-kDa whereas 1F3 clone with two proteins, 40- and 45-kDa. Their isotypes against mouse Ig family were identical, kappa-light chain. These results suggest that these monoclonal antibodies can be useful for the development of diagnostic kit for the pine wilt disease.

Sex pheromone production in lepidopteran is stimulated and regulated by a pheromone biosynthesis activating neuropeptide (PBAN). A cDNA of PBAN receptor (Plx-PBANR) isolated from female pheromone gland of the diamondback moth (DBM, Plutella xylostella (L.) encodes 338 amino acids. Plx-PBANR has conserved biochemical motifs and 7 transmembranes, indicating it belongs to G-protein coupled receptor family. Plx-PBANR expression was only found in female pheromone gland, demonstrating that pheromone gland is the only molecular target of Plx-PBAN. Human uterus carcinoma (HeLa) was stably transfected with Plx-PBANR gene and its expression was confirmed by RT-PCR analysis. Plx-PBANR expressing cells increased level of Ca2+ influx when challenged with Plx-PBAN and Hez-PBAN from Heliothis zea. When RNAi fragment for PBANR was injected into pupae, suppression of PBANR expression was confirmed by RT-PCR and maintained for at least 2 days at post-emergence. Injection of RNA fragment into pupae for inhibition of Plx-PBANR expression also inhibited mating behavior, revealing that reproductive organ of the female has no spermatocyte and that there are no successful reproductive behaviors. These results suggest of PBANR expression affects the molecular biological events of PBAN and eventually suppresses mating behavior.

The Black Soldier Fly (BSF, Hermetia illucens) was widely distributed throughout Korea. This insect was mainly found in the vicinity of and in cattle sheds, manure sheds, living waste dump grounds, and food waste dump grounds. This fly is a kind of a beneficial fly because BSF adults do not go into houses, they do not regurgitate on human food, they do not bite, bother or pester humans in any way and they are not associated in any way with the transmission of disease. But their greatest attribute lies their ability to eat and digest raw waste. They can devour, for example, a large, raw, Irish potato and others in just a few hours. Unlike many other flies, since the BSF larvae have very powerful mouth parts and digestive enzymes, they can ingest raw waste far more efficiently than any other known species of fly. On this study, to investigate whether feeding strategy of the BSF larvae involves extra-oral digestion or not, and to better understand this process, the salivary glands and a few tissue from the BSF were dissected and subjected to morphological and preliminary enzyme characterization.

Varroa destructor is an ecto-parasite mite and worldwide pest of the honey bee Apis mellifera L. The pyrethroid tau-fluvalinate (Apistan), an acaricide that is tolerated by honey bees, has been used for varroa mite control since the mid 1980s. Even though various resistances to tau-fluvalinate in varroa mites have been reported from Europe, Israel, and USA, the nature of tau-fluvalinate resistance in varroa mites in Korea has never been investigated. To investigate and understand tau-fluvalinate resistance in varroa mites in Korea, we conducted bioassay in several apiaries located different regions in Korea. To understand the molecular mechanisms underlying the difference of tau-fluvalinate resistances in varroa mites, partial genomic DNA fragments of a voltage-sensitive sodium channel gene from varroa mites were cloned and sequenced, since tau-fluvalinate is known to act on the sodium channels directly. Two novel mutations in sodium channels of varroa mites were present in eight apiaries. Two mutations might be a geographical polymorphism of sodium channel of varroa mites in Korea.

A cDNA of PBAN receptor (Plx-PBANR) isolated from female pheromone gland of the diamondback moth (DBM, Plutella xylostella (L.) encodes 338 amino acids. Plx-PBANR includes 7 transmembranes, indicating it belongs to G-protein coupled receptor family. Plx-PBANR showed high similarities with other moth PBANRs and its expression was only found in female pheromone gland, demonstrating that pheromone gland is the only molecular target of Plx-PBAN. To accomplish the funcional expression of Plx-PBANR, Human uterus carcinoma was stably transfected with Plx-PBANR gene and Plx-PBANR expression was confirmed by RT-PCR analysis. Plx-PBANR expressing cells increased level of Ca2+ influx when challenged with Plx-PBAN and Hez-PBAN from Heliothis zea, as ionomycin as a positive control does. To inhibit Plx-PBNAR expression in vivo, RNAi fragment for Plx-PBANR was injected into pupae. Suppression of PBANR expression was confirmed by RT-PCR and also induced inhibition of mating behavior in adults, revealing that reproductive organ of the female has no spermatocyte and that there are no successful reproductive behaviors. RNAi-treated adults showed reduced pheromone production. These results suggests that inhibition of PBANR expression affects the molecular biological events of PBAN and eventually suppresses mating behavior.

The pine sawyer beetle, Monochamus alternatus, transmits the pinewood nematode (PWN, Bursaphelenchus xylophilus), causing the pine wilt disease (PWD), which gives rise to enormously economic as well as forest damage. However, PWN has been identified as a pathogen of PWD, it is very difficult to discriminate B. xylophilus from B. mucronatus in a short time, which are genetically and morphologically very similar. Therefore, it has been necessary to detect and eliminate PWN-infected trees in the forest area for the prevention of PWD transmission. Up to date, there is no report on biomarkers such as DNA and protein for the diagnosis of B. xylophilus. In this study, we produced a B. xylophilus monoclonal antiserum (D9-F10) from BalbC mice and screened its specificity with various proteins extracts. Western blot analysis revealed that the D9-F10 is only reactive with B. xylophilus protein extract among other tested protein extracts, indicating that the D9-F10 is specific for a B. xylophilus protein. Furthermore, two-dimensional electrophoresis showed the D9-F10 detects a very highly acidic protein, pI≒3.5. These results suggest that the D9-F10 monoclonal antibody is useful for the development of a diagnostic kit for the pine wilt disease.

The pinewood nematode (PWN, Bursaphelenchus xylophilus) causes the pine wilt disease, transmitted to pinewoods by the pine sawyer beetle, Monochamus alternatus. It is very difficult to discriminate B. xylophilus from B. mucronatus. Therefore, it has been necessary to detect PWN-infected trees for the prevention of pine wilt disease transmission in a short time. The development of biomarkers such as DNA and protein is important for diagnosis of B. xylophilus. However, there have been no reports regarding biomarker identifications of B. xylophilus. In this study, polyclonal antisera were raised against whole proteins of B. xylophilus in BalbC mice and were primarily screened with ELISA. Twenty five among over 500 cell lines releasing polyclonal antisera revealed B. xylophilus-specific immuno-reactivity, which was at least three times higher than that of B. mucronatus. Three cell lines among them were secreting monoclonal antibody through further screening. These cell lines only detect about a 33-kDa protein in B. xylophilus in the western blot. These results suggest that these monoclonal antibodies will be useful for the development of diagnostic kit for the pine wilt disease.

Many female moths produce and release sex pheromones to mate successfully with a conspecific male. Sex pheromone production in lepidopteran moth is known to be under the regulation of a pheromone biosynthesis activating neuropeptide (PBAN). A PBAN polypeptide is processed into five neuropeptides (NPs) after post-translational modification, resulting in diapause-like NP, PBAN, α-, β- and γ-NP. All of the peptides are amidated in their C-termini and shared a conserved motif, FXPR(or K)L structure. PBAN (Plx-PBAN) from Plutella xylostella consists of 30 amino acids, the shortest PBAN so far reported. When female adults were injected with synthetic Plx-PBAN, pheromone production showed a maximal increase 1 h post-injection. RT-PCR screening revealed that Plx-PBAN cDNA was expressed in both sexes, with the highest expression level in the head of female adults. Plx-PBAN binds to its receptor on pheromone gland cells. PBAN receptor has seven transmembranes, indicating G-protein coupled receptor family and transduces its signal via G-protein mediated signal transduction. Subsequently, calcium channels remain activated and stimulate biochemical reactions for sex pheromone production in the pheromone gland.

인체피부에서 화장품이나 화장품 원료의 안전성시험은 대체로 육안평가로 이루어져왔다. 하지만, 피부반응 초기의 미세한 변화에 대해서 육안으로는 감지하지 못하는 경우가 많다. 따라서, 본 연구에서는 수종의 화장품 원료에 의해 유발되는 홍반반응에 대해 육안평가뿐 아니라, 레이저도플러혈류이미지(LDPI) 측정장비를 이용한 혈류변화 측정, Vapometer®를 이용한 경피수분손실량(TEWL) 측정, 분광광도계를 이용한 피부색 측정과 같은 기기평가를 병행하여 비교하였다. 30명의 건강한 여성 피험자를 대상으로 오일류 7종, 계면활성제류 6종, 보습제류 5종을 피험자의 등 부위에 24 h 폐쇄첩포(D0)하여 피부반응을 유도하였고 첩포 제거 후 30 min (D1), 24 h (D2)에 육안평가와 기기평가를 각각 실시하였다. 육안평가 결과, 프로필렌글라이콜을 제외한 모든 오일류와 보습제류는 저자극 수준의 피부반응(반응도 0+ ~ 2.9+)을 보인 반면, 프로필렌글라이콜과 모든 계면활성제류는 중자극에서 강자극 수준의 피부반응(반응도 3+ ~ 5+)을 보였다. 기기측정 결과, 육안평가에서 저자극 범주의 피부반응에 대해서 혈류량이나 피부색보다 경피수분손실량이 가장 민감하게 피부변화를 감지하였다.

This study was conducted to identify QTLs for some agronomic traits under two different N-fertilizer conditions. A recombinant inbred population (DT-RILs) between Dasanbyeo (a tongil type rice, derived from an indica × japonica cross and similar to indica in its genetic make-up) and TR22183 (a Chinese japonica variety), consisting of 166 F8 lines, was developed and used for mapping. A frame map of 1,409 cM containing 113 SSR and 103 STS markers with an average interval of 6.5 cM between adjacent marker loci was constructed using the DT-RILs. The DT-RILs were cultivated in ordinary N-fertilizer (N-P2O5-K2O=100-80-80 kg/ha) condition and low N-fertilizer (N-P2O5-K2O=50-80-80 kg/ha) condition in order to compare the different N-fertilizer effects. Grain yield was positively correlated with spikelet number in both conditions. Path analysis revealed that spikelet number is the most positive contributor to grain yield. Twenty-nine single QTLs (S-QTLs) and sixty-five pairs of epistatic loci (E-QTLs) were identified for days to heading, culm length, panicle length, panicle number, spikelet number, spikelet fertility, 1000-grains weight and grain yield in both N-fertilizer conditions. The phenotypic variance explained by these S-QTLs and E-QTLs ranged from 8.8% to 61.1% and from 10.0% to 55.6%, respectively. The total phenotypic variance explained by all the QTLs for each trait ranged from 15.4% to 92.3%.