Although it is believed that internal nutrient sensors play important roles in feeding behaviors, their molecular and neural mechanisms underlying of the modulation of physiological status and cell growth are poorly understood. Using a Ca2+ imaging experiments with heterologous expression systems, we show that one of the gustatory receptors in the western honey bee Apis mellifera is selectively tuned to amino acids. Remarkably, we report that this gustatory receptor of the honey bee is highly expressed in hypopharyngeal gland, which plays a role in caste differentiation as well as royal jelly production and secretion. Knocking down this gustatory receptor gene reduces cellular pathways responsible for nutritional sensing such as mTOR signals in hypopharageal gland. Furthermore, the interfering expression of this gustatory receptor gene not only alters morphological changes and developmental retardation of the hypopharyngeal gland, but it also blocks cellular growth signals to induce autophagy. This new report indicates that internal sensing and downstream signals detecting nutrients is essential for honey bee to maintain the cellular growth and development of internal organs essential for caste development and maintenance of social structure in the honey bee.

Multiple starters consisting of two Bacillus amyloliquefaciens strains (MJ1-4 and EMD17), Pichiafarinosa SY80, and Rhizopus oryzae were used for Doenjang making. Bacillus strains were selected based on their abilities to inhibit toxinogenic fungi and Bacillus cereus, fibrinolytic activity, and their ability to confer good flavor to Cheonggukjang. P. farinosa SY80 and R. oryzae, previously isolated from soy sauce, were selected because they were not inhibited by two bacilli. Doenjang was prepared by inoculation of multiple starters (A1 Doenjang). Control Doenjang was prepared by inoculation of B. subtilis KACC 16750 (Natto strain) and Aspergillus oryzae KCCM 60166 (A2 Doenjang). Another control (A3 Doenjang) was prepared by inoculation of microorganisms present in rice straw. Doenjang samples were fermented for 70 days at 20℃. pH of 3 samples decreased from the initial value of 6.4 to 5.8~6.0 and titratable acidity (TA) increased from 0.6 to 1.1~1.3. The amount of amino-type nitrogen increased during fermentation. There were slight differences in moisture, crude-protein, and crude-fat contents after 70 days. Contamination of fungi was observed only in A3 Doenjang and B. cereus was not detected from all 3 samples. A1 Doenjang showed the highest fibrinolytic activity and A2 Doenjang the second. These results indicate that Doenjang made with carefully selected starters was functionally improved and microbially more safe.

In the present study, lichen(Parmelia sp.) extract showed insecticidal activity against Aedes albopictus, and the effect of growth inhibiting activity was investigated. Acetone and methanol extracts of the lichen against larvae of Aedes albopictus were showed high insecticidal activities in low lethal concentration. 50% lethal concentration of the acetone extractof the lichen is 0.13% and the 50% lethal concentration of methanol extract of lichen is 0.15% respectively. This experiment that used acetone and methanol extracts of the lichen were observed for 24, 48 and 72 hours. As a result, the higher concentration and the longer exposure time is increased mortality against Aedes albopictus. Pupation time took more time as the higher concentration of acetone extract of lichen. Consequently, the lichen extract is effect in inhibiting the growth of Aedes albopictus larvae. In this experiment indicates that lichen extract has activity against Aedes albopictus and is available as the natural insecticide.

Inhibitor cysteine knot (ICK) peptides exhibit ion channel blocking, insecticidal, and antimicrobial activities, but currently, no functional roles for bee-derived ICK peptides have been identified. In this study, a bee (Apis cerana) ICK peptide (AcICK) that acts as an antifungal peptide and as an insecticidal venom toxin was identified. AcICK contains an ICK fold that is expressed in the epidermis, fat body, or venom gland and is present as a 6.6-kDa peptide in bee venom. Recombinant AcICK peptide (expressed in baculovirus-infected insect cells) bound directly to Beauveria bassiana and Fusarium graminearum, but not to Escherichia coli or Bacillus thuringiensis. Consistent with these findings, AcICK showed antifungal activity, indicating that AcICK acts as an antifungal peptide. Furthermore, AcICK expression is induced in the fat body and epidermis after injection with B. bassiana. These results provide insight into the role of AcICK during the innate immune response following fungal infection. Additionally, we show that AcICK has insecticidal activity. Our results demonstrate a functional role for AcICK in bees: AcICK acts as an antifungal peptide in innate immune reactions in the body and as an insecticidal toxin in venom. The finding that the AcICK peptide functions with different mechanisms of action in the body and in venom highlights the two-pronged strategy that is possible with the bee ICK peptide.

Cotesia plutellae known as an endoparasitoid parasitizes larvae of the diamondback moth, Plutella xylostella which is a major pest in cruciferous crops. For the successful parasitization, maternal and embryonic factors of C. plutellae such as polydnavirus, ovarian proteins, teratocytes and venom are required. In this study, we identified calreticulin (Cp-CRT) gene from transcriptome data of the venom gland in C. plutellae. cDNA of CRT was cloned from total RNA of the venom gland via PCR and encodes 403 amino acids harboring several structural motifs such as a signal peptide sequence, a repetitive sequence, a putative coiled-coil sequence encompassing, and endoplasmic reticulum-recognizing domain (-KDEL). Phylogenetic analysis showed that the Cp-CRT gene formed a unique cluster with other hymenopteran CRT genes, indicating that the Cp-CRT belongs to the CRT family. To examine the physiological function of Cp-CRT, recombinant Cp-CRT, fused with 6X-His at N-terminal was constructed and expressed in E. coli. Recombinant Cp-CRT was successfully expressed via Western blot analysis and suppressed significant nodule formation when co-injected with E. coli as immune response inducer. These results suggest that the Cp-CRT involves in suppression of cellular immune response in the host

Symbiotic bacteria are common in insects. Because symbiotic bacteria are known to intimately affect the various aspects of insect host biology, ideally insects can be controlled by manipulating their symbiont. However, the attempts to control insects through their symbiont have been very limited. The paucity of the insect pest control using their symbiont is most likely due to the poor understanding of the symbiotic interactions between host insect and symbiont, which is attributed to the difficulty in cultivation of insect symbionts. However, the recently established bean bug, Riptortus pedestris, symbiotic system provides good opportunities to study insect’s symbiont in molecular level through their cultivable symbionts. Bean bugs acquire genus Burkholderia cells from environment and harbor them as their gut symbionts in the specialized posterior midgut. The genome of the Burkholderia symbiont was sequenced, and the genomic information has been used to generate the genetically manipulated Burkholderia symbiont strains. After orally administering the mutant Burkholderia symbionts into bean bugs for symbiotic association, the bacterial colonization levels in the host gut and host phenotypes were analyzed. As a result, we have identified novel symbiotic factors necessary for establishing successful association with host. Our recent understandings on the bacterial symbiotic factors demonstrate a great possibility to control the bean bug pest using genetically modified Burkholderia symbiont.

Germplasm collection of lily in the mountainousarea is laborious and time consuming accompanied withhigh costs and high risk due to inappropriate environmentalconditions. Cryopreservation being an ideal method for thelong-term preservation can be employed in conservation ofvaluable lily germplasm. Previously, we developed cryo-preservation protocol for Lilium germplasm using ‘dropletvitrification’. In this paper, we have chosen shoot tips as amaterial for cryopreservation because of their genetic safetyupon regrowth in tissue culture. Using this protocol, wehave preserved approximately 160 accessions of lily germ-plasm in 2010~2012. The regeneration rates are rangedfrom 54.3% to 58.5% while the survival rates were from58.3% to 66.4%. Among Lilium germplasm cryopreserved,there are some Korean, Chinese, and Taiwanese seedstocks which have good qualities for inter-species hybrid.Moreover, we also conserved Korean wild endangered seedstock, especially Lilium hansonii. The morphological studyof Lilium germplasm regenerated from cryopreserved mate-rial confirmed the stability of clonal material following cryo-preservation. We anticipate this cryo-collection will beavailable and useful to curators or breeders of Lilium andthis cryobank will also facilitate the conservation and inter-national exchange of Lilium germplasm.

This study examines the determinants of the member customer’s decision of redeeming versus accumulating loyalty program (LP) points by focusing on the effects of the different channels of transaction (online versus offline) and the demographic information of member customers. Our study is based on customer-level demographic and transaction data on a major partnership LP in Korea, the OK Cashbag (OCB) program. This study differs from the existing literature in three aspects. First, the dataset employed for this study enables us to compare member customers’ point redemption behavior between online and offline channels, whereas previous studies demonstrate coupon redemption behavior either in an online (Chiou-Wei and Inman 2008) or an offline setting (e.g., Cronovich 1997; Kwon and Kwon 2007; Mittal 1994; Reibstein and Traver 1982; Ward and Davis 1978). Second, the current study investigates not only the main effects of demographic variables, but also a series of interaction effects between the online channel and each demographic variable. Clear empirical evidence of an interaction effect would provide an LP provider with significant managerial implications. Third, rich data on customers’ transaction behavior with matching demographic information for each member customer enable us to conduct both transaction-level and individual customer-level analyses. Therefore, an individual customer’s transaction behavior can be analyzed in more detail for robust results and richer implications. We find that transactions that occur through online channels and those made by younger customers demonstrate a greater tendency of redeeming LP points as opposed to accumulating them. We also find that online channels exhibit a moderating role by mitigating the demographic effects on member customers’ point redemption behavior. These findings allow the LP provider to predict the future LP point balance by analyzing its main channel of transaction and the demographic profiles of its member customers.

The purpose of this research is to initiate discussion to simplify existing tools and methods for managing projects which make project management a harder job to perform and a tougher task to handle. Additionally, summarizing the definitions of “project” that have been found from reviewing the relevant literature, it deliberates on revising the duties and responsibilities of project manager and proposes performance expectancy triangle explaining a relationship among accountability, responsibility and authority – the Tri-butes. It sheds light on several factors critical to the success of projects. Finally, we propose a function indicating that customer demands or desires greatly affect project complexity.

The Riptortus (stinkbug) has a specialized symbiotic organ, M4 midgut, to harboring symbiont Burkholderia. M4 midgut is located in abdomen and surrounded with insect hemolymph. Recently our group demonstrated that symbiotic Burkholderia showed different physiology after adapting in M4 gut compare with in vitro cultured Burkholderia. And population of symbiotic Burkholderia in the M4 midgut is regulated by special organ. However, the molecular mechanism to prevent spreading and migrating symbiont bacteria to other host tissues from symbiotic organ is not clear. Therefore, we assumed that symbiont Burkholderia are susceptible to host humoral immunity after established infection in M4 midgut to prevent spreading and migrating into the other host tissues through Riptortus hemolymph. To prove this assuming, we tested the susceptibility and survival rate of symbiont Burkholderia in hemolymph of Riptortus in vitro and in vivo. We also examined the susceptibility of symbiont Burkholderia using purified antimicrobial peptides (AMP), pyrrhocoricin-like, thanatin-like and defensin-like AMPs. Finally, we tested inducing ability for AMPs by systemic infection of symbiotic Burkholderia. Gene expression of purified AMPs was not different after systemic infection of both symbiont and in vitro cultured Burkholderia. Surprisingly, in vitro cultured Burkholderia resisted on bacteria injected hemolymph and purified AMPs but symbiont Burkholderia were highly susceptible in bacteria injected hemolymph and purified AMP. These results suggest that symbiont Burkholderia can't survive in the hemolymph after escaping symbiotic organ. Moreover, humoral immunity of host Riptortus is important to prevent spreading and migrating symbiont Burkholderia into the other host tissue or organ from symbiotic organ.

Cotesia plutellae as an endoparasitoid wasp is the natural enemy against the diamondback moth, Plutella xylostella. For successful parasitism of the offspring, the female parasitoid always injects venom with other parasitic factors such as polydnavirus, teratocytes and ovary proteins. These venom products prevent their eggs from host immune system as well as modulate their host. To identify of constituents of venom in relation of host-parasitoid interaction, total RNA of the venom gland were extracted and analyzed with RNA-seq. RNA-seq reveals that 5.5 Gbp were read and approximately 3322 transcripts were identified. The transcripts related to the biological process, cellular components and molecular functions occupied about 20%, 15% and 18%, respectively. Among these transcripts, 301 genes were related to the immune activities such as calreticulin, defensing, phosphatases and serpin etc. These results suggest that some proteins exhibiting biological function related to the immune response can be contribute to the development of wasp in the host.

Olfactory sensitivity exhibits daily fluctuations. Several studies have suggested that the olfactory system in insects is modulated by both biogenic amines and neuropeptides. However, molecular and neural mechanisms underlying olfactory modulation in the periphery remain unclear since neuronal circuits regulating olfactory sensitivity have not been identified. Here, we investigated the structure and function of these signaling pathways in the peripheral olfactory system of the American cockroach, Periplaneta americana, utilizing in situ hybridization, qRT-PCR, and electrophysiological approaches. We showed that tachykinin was co-localized with the octopamine receptor in antennal neurons located near the antennal nerves. In addition, the tachykinin receptor was found to be expressed in most of the olfactory receptor neurons in antennae. Functionally, the effects direct injection of tachykinin peptides, dsRNAs of tachykinin, tachykinin receptors, and octopamine receptors provided further support for the view that both octopamine and tachykinin modulate olfactory sensitivity. Taken together, these findings demonstrated that octopamine and tachykinin in antennal neurons are olfactory regulators in the periphery. We propose here the hypothesis that octopamine released from neurons in the brain regulates the release of tachykinin from the octopamine receptor neurons in antennae, which in turn modulates the olfactory sensitivity of olfactory receptor neurons, which house tachykinin receptors.

Insect-derived Kazal-type serine protease inhibitors exhibit thrombin, elastase, plasmin, proteinase K, or subtilisin A inhibition activity, but so far, no functional roles for bee-derived Kazal-type serine protease inhibitors have been identified. In this study, a bee (Apis cerana) venom Kazal-type serine protease inhibitor (AcKTSPI) that acts as a microbial serine protease inhibitor was identified. AcKTSPI contained a single Kazal domain that displayed six conserved cysteine residues and a P1 threonine residue. AcKTSPI was expressed in the venom gland and was present as a 10-kDa peptide in bee venom. Recombinant AcKTSPI Kazal domain (AcKTSPI-Kd) expressed in baculovirus-infected insect cells demonstrated inhibitory activity against subtilisin A (Ki 67.03 nM) and proteinase K (Ki 91.53 nM), but not against α-chymotrypsin or typsin, which implies a role for AcKTSPI as a microbial serine protease inhibitor. However, AcKTSPI-Kd exhibited no detectable inhibitory effects on factor Xa, thrombin, tissue plasminogen activator, or elastase. Additionally, AcKTSPI-Kd bound directly to Bacillus subtilis, B. thuringiensis, Beauveria bassiana, and Fusarium graminearum but not to Escherichia coli. Consistent with these findings, AcKTSPI-Kd showed antibacterial activity against Gram-positive bacteria and antifungal activity against both plant-pathogenic and entomopathogenic fungi. These findings constitute molecular evidence that AcKTSPI acts as an inhibitor of microbial serine proteases. This paper provides a novel view of the antimicrobial functions of a bee venom Kazal-type serine protease inhibitor.

Cotesia plutellae has been known as a natural enemy against the Diamondback moth, Plutella xylostella via laying eggs into a larva. When the larva hatches from the egg, teratocytes also are released and expected to work as immune suppressor via secreting immune suppressive factors such as venom proteins, teratocytes and polydnavirus. In order to identify immune suppressive factors from teratocytes, we collected the supernatant from serum-free culture media of teratocytes. Concentration of secreted proteins from teratocytes was successfully performed with using centricons among tested methods such proteins precipitation and electrophoresed in sodium dodecyl sulfate polyacrylamide gel. The gel slices were directly digested with trypsin using in-gel digestion method and analyzed via LC-Ms-Ms. Molecular weight of peptide fragments were compared with protein database predicted by full-genome sequences of C. plutellae. We identified two immune responsive proteins, which are calreticulin, host cellular response-related gene and neprilysin 2, immune regulator. This result suggests that host immune response is suppressed or regulated by the immune suppressive factors of teratocytes.

소듐 2-브로모에탄술포네이트를 이용한 2-에티닐피리딘의 무촉매중합을 통하여 새로운 이온성 폴리아세틸렌을 합성하였다. 중합반응은 균일하게 진행되었으며 높은 수율 (중합수율: 78%)로 해당고분자를 합성하였다. 반응 초기 사차 염화과정에서 생성된 활성화 N-(에틸술포네이트 소듐)-2-에티닐피리디늄 브로마이드가 중합반응의 단량체로 참여하고 있는 것으로 밝혀졌다. 고분자의 분자 구조를 여러 가지 분석기기로 확인한 결과 설계한 치환기를 갖는 폴리아세틸렌 유도체가 합성되었음을 확인할 수 있었다. 합성 고분자의 고유점도는 0.12-0.15 dL/g 범위였으며 X-선 회절분석 결과 무정형상임을 확인할 수 있었다. 고분자의 광발광 피크가 593 nm에서 관찰되었는데, 이는 2.09 eV의 광에너지에 해당한다. 이 고분자는 도핑과 탈도핑 피크 사이에서 비가역 전기화학적 거동을 보였다.

선충절대기생세균(Pasteuria penetrans)이 감염되어 있는 온실토양을 이용하여 세균의 내생포자가 땅콩뿌리혹선충(Meloidogyne arenaria) 유충 표면에 부착하는데 대한 온도의 영향에 대해 시험하였다. 갓 부화된 뿌리혹선충 2령충(J2)을 페트리디쉬 내의 토양에 접종한 후 20℃, 25℃, 30℃, 35℃에서 7일간 처리하였다. 모든 온도에서 내생포자의 J2 부착률은 모두 100%로 나타났으나 J2당 내생포자 부착수는 25℃에서 28.3개로 가장 많았으며 30℃, 20℃ 및 35℃에서 각각 J2 당 20.2, 18.6 및 13.6개로 낮아졌다. J2를 접종하기 전에 세균이 있는 토양을 온도 별로 10일간 전처리하였을 때 내생포자 부착률은 실온에서의 60%에 비해 -30℃, 4℃, 40℃, 50℃ 및 100℃에서 각각 25.0, 31.7, 8.3, 5.0 및 0%로 현저하게 낮아졌다. J2 당 내생포자 부착수도 실온에서의 5.3개에 비해 -30℃, 4℃, 40℃, 50℃ 및 100℃에서 각각 3.5, 4.3, 1, 1, 0개로 적었다. P. penetrans 세균의 내생포자를 뿌리혹선충 종별로 J2에 부착 시험한 결과 땅콩뿌리혹선충에서는 100%였으나 당근뿌리혹선충(M. hapla)과 고구마뿌리혹선충(M. incognita)에서는 모두 0%로 본 균주는 뿌리혹선충 종에 대해 기주선호성을 가진 것으로 나타났다.