Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.
It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.
IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.
본 연구는 기존에 재배된 바 없었던 암대극을 새로운 관상식물로 개발하고자, 절화 특성 평가를 실시하였다. 2017년 3월과 4월, 제주도에 자생하고 있는 암대극을 채취한 후, 수분흡수, 절화수명, 노화 과정 등 절화수명 특성을 평가하였다. 또한 sucrose 농도, 절화길이, pH 농도, 저장온도에 따른 절화 특성을 조사하였다. 본 실험은 국립수목원 유용식물증식센터의 Phyto-garden system(12h photoperiod, 25℃, 70% RH)과 실온(23℃, 44% RH)에서 수행하였고 2~3일 간격으로 수분흡수량, 생체중, 절화수명을 측정하였다. 실험 환경에 따른 연구에서 실온(14.4일) 보다 phyto-garden system(42.4일)의 절화 수명이 약 3배 더 길었고, 절화길이에 따른 연구에서는 절화수명이 20cm, 15.2일, 40cm, 17.4일로 나타났다. Sucrose 농도에 따른 연구에서는 처리에 따른 절화수명 증진 효과는 없었고, pH에 따른 연구의 절화수명은 pH 5, 15.6일, pH 6, 13.4일, pH 7, 13.1일, 증류수, 14.8일이었다. 저장온도에 따른 실험에서, 절화수명은 4℃, 83일, 10℃, 41.2일, 15℃, 35.5 일, 20℃, 17.4일, 실온, 14.4일이었는데, 온도가 감소할수록 절화수명이 길어짐을 알 수 있었다. 결론적으로, 암대극은 비교적 광도와 습도가 높은 환경과 절화길이가 길고 저장온도가 낮을수록 수명이 오래 유지되는 것을 알 수 있었다.
본 연구는 딸기에서 발생하는 흰가루병 방제에 사용되는 살균제의 잔류시험을 수행하고 회귀방정 식 및 생물학적 반감기를 산출하여 한국과 일본, Codex의 잔류허용기준에 근거한 농약의 사용 및 안전한 농산물 생산에 기여하고자 수행되었다. 딸기재배 중 흰가루병 방제용 주요 살균제 boscalid, kresoxim-methyl, pyraclostrobin, pyrimethanil을 각 농약의 안전사용기준에 따라 기준량 1회 살포 후 1, 3, 5, 7, 9, 11, 13일에 딸기 시료를 채취하여 QuEChERS법을 이용하여 추출하고, NH2 SPE 카 트리지로 정제하여 HPLC/DAD로 분석하였다. 시험농약의 검출한계 및 정량한계는 4성분의 농약에서 모두 0.01과 0.03mg/kg이었으며, 평균회수율은 92.9~99.2%, 상대표준편차는≤5%로 산출되었다. 딸기 중 boscalid, kresoxim-methyl, pyraclostrobin, pyrimethanil의 생물학적 반감기는 기준량처리 시 각각 6.2, 4.0, 6.1, 6.6일로 나타났다. 이러한 결과로, 시설재배지 딸기 흰가루병 방제를 위해 사용되는 살균제의 안전 사용을 통해 국내 및 주요 수출국가의 MRL을 초과하지 않는 안전한 딸기를 생산하는데 기여할 것으로 기대된다.
기체 분리막 소재에 있어서, 자유체적 및 기계적 특성은 기체투과 성능 및 소재 응용에 있어서 필수적인 요소 들이다. 따라서, 기체 투과 성능의 핵심인 자유체적 비율을 최대한 높이면서도, 동시에 높은 연신율을 가질 수 있는 고분자 구조의 설계가 필수적이며, 이를 위해서는 각각의 특성을 분자 수준에서 이해하고 분석을 할 수 있어야 한다. 본 연구에서는 분자전산모사(molecular simulation) 기술을 이용하여, 기존에 실험적으로 밝혀진 TR-polymer와 spiro-group이 도입된 TR-PBO 간의 차이를 분자수준에서 밝혀냄으로서, 고분자 구조내의 특정 작용기가 최종 고분자 소재의 자유체적 및 기계적 특성에 어떻게 작용하는지를 규명하고자 한다.
Several species of the genus Aphidius are used in biological control programs against aphid pests throughout the world and their behavior and physiology are well studied. But despite knowing the importance of sensory organs in their behavior, their antennal structure is largely unknown. In this study, the external morphology and distribution of the antennal sensilla on the antennal of both female and male adults of A. colemani were described using scanning electron microscopy (SEM). Generally, the filaform antennae of males (1,515.20±116.48 ㎛) are longer than females (1,275.06±116.42㎛). Antennae of this species is made up of scape, pedicel and flagellomeres. Male and female antennae differed in the total number of flagellomeres as 15 in males and 13 in females. Female and male antennae of A. colemani has samely seven types of sensilla. We classified sensilla placodea, Bohm bristles, 2 types of sensilla coeloconica, , 2 types of sensilla basiconica as with a tip pore and with wall pores, sensilla trichodea. In addition, the possible functions of the above sensilla types are discussed in light of previously published literature; mechanoreception(Bohm bristles, sensilla coeloconicaⅡ and sensilla trichodea) and chemoreception(sensilla coeloconicaⅠ, sensilla basiconicaⅠ,Ⅱ and sensilla placodea). Future studies on the functional morphology of the antennal sensilla of A. colemani using transmission electron microscopy (TEM) coupled with electrophysiological recordings will likely confirm the functions of the different sensilla identified in this study.
Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, H+-transporting ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.
Background : Undulatum Rhubarb, commonly produced in domestic, is rhizome of Rheum undulatum L. that belongs to the family Polygonaceae. It also can be used as a substitute of R. palmatum L., R. tanguticum Maximowicz ex Balf., and R. officinale Baillon which completely depend on import system. However, there should be clear clarification among Undulatum Rhubarb and Rhubarb, because Undulatum Rhubarb contains rhaponticin as marker compound that is not indicated at Rhubarb. Some of the recently imported Undulatum Rhubarbs have been found to be Rhubarb. Also, it is known that only Undulatum Rhubarb is cultivated at domestic environment. But some plant origins of Rhubarb are grown in Korea, too. Further study are needed to clarify clear origin between Undulatum Rhubarb and Rhubarb. Thus, we collected some domestically cultivated samples and identified them. Methods and Reseults : Rheum undulatum L., Rhubarb, Rheum tanguticum Maximowicz ex Balf. which were cultivated in Gangwondo Agricultural Research and Extension Services in Cheorwon were collected and anayzed the DNA sequences. We also compared DNA sequences in Rhubarb collected from England and R. rhabarbarum L., R. undulatum L., and R. franzenbachii on NCBI. As a result, two kinds of rhubarb cultivated in the test plantation were identified as R. rhabarbarum and R. officinale. In addition, R. undulatum (plant origin of Undulatum Rhubarb) was identified as Rhubarb (Rheum rhabarbarum) in England with 99 - 100% identical in nuclear ITS gene region. Conclusion : R. undulatum, plant origin of Undulatum Rhubarb, is reported as synonym of R. rhabarbarum, R. franzenbachii. Rheum speices which are cultivated as tester in Gangwondo Agricultural Research and Extension Services in Cheorwon are estimated as R. undulatum and R. officinale. Therefore, not only Undulatum Rhubarb but Rhubarb could be grown in Korea.
Background : Perilla frutescens L. is valuable as a medicinal plant as well as a natural medicine and functional food. Limonene perilla collected from various places showed 60% limonene compounds. However biological activity of these accession has not been reported before. Therefore, this study was conducted to investigate the biological activity of limonene perilla. Methods and Results : Fractional solvent extracts were obtained by using organic solvents such as n-hexane, chloroform, ethyl acetate, n-BuOH, and aqueous solvent from different parts of limonene perilla extracted initially in 70% EtOH. We investigated the effects of limonene perilla on total phenol and flavonoid contents, FRAP (Ferric Reducing Antioxidant Power), total saponin contents and tyrosinase inhibition activity. Leaves of limonene perilla produced the highest total phenolic contents (29.88 mg·CAE/g), flavonoid (8.39 mg·QE/g) and saponin contents (47.77 mg·GIE/g) than stems and roots of limonene perilla. FRAP of leaves was 823.00±3.58 μM·FeSO4·E/mg. Tyrosinase inhibition activity rate was 40.31% in 70% ethanol extracts from leaves of limonene perilla. Conclusion : This results suggest that leaf of limonene perilla fractions has significant antioxidant activity. Also, limonene perilla could be used as a functional biomaterial in developing cosmetics and functional foods.