Pine wilt is the most important disease of pine trees in Korea, Japan and China. The pathogen causing this disease, the pinewood nematode (Bursaphelenchus xylophylus), is transmitted vectored by adults of some cerambycid beetle species and the Japanese pine sawyer, Monochamus alternatus, is the major vector species in Korea. Although chemical insecticides have been used to kill vector insect and thus prevent transmission of the pathogen, the efficacy is not good. In Japan, to control this insect, an entomopathogenic fungus was studied and developed as an insecticide. This is thought to be the convenient and effective method to control M. alternatus. Recently, there are several reports about the pinewood nematode is vectored by also the pine sawyer, M. saltuarius, in Korea. The objective of this study, therefore, was to isolate and identify entomopathogenic fungi from M. saltuarius cadaver to control it. We collected the cadaver of M. saltuarius and then screened several fungi colonies. The pathogenicity of each fungus was tested using oak longicorn beetle, Moechotypa diphysis, as substitutive insect. M. diphysis is also serious pest to various trees in forest. As the result, only one of them showed high pathogenicity against M. diphysis. Selected fungus was identified by microscopic examination and DNA analysis. Pathogenicity was also evaluated to M. saltuarius.

폴리스티렌-블록-폴리히드록에틸 메타크릴레이트(PS-b-PHEMA), 술포석시닉산(SA), 인텅스텐산(PWA)으로 구성된 수소 이온 전도성 나노복합 고분자 전해질막을 제조하였다. 폴리히드록에틸 메타크릴레이트(PHEMA) 블록의 히드록실그룹(-OH)와 술포석시닉산(SA)의 -COOH 그룹과의 에스테르 반응에 의하여 전해질막을 가교시켰다. 폴리헤테로산(PWA)을 도입했을 때, SO3 - 그룹의 신축 밴드가 1187 cm -1에서 1158 cm -1로 낮아졌으며, 이는 PWA 입자가 전해질막의 술폰산 그룹과 상호작용함을 나타낸다. PWA 함량이 30wt%가 되었을 때, 상온 전도도는 0.045에서 0.062S/cm로 증가되었으며, 이는 PWA 입자의 고유 전도도 특성과 전해질막의 술폰산기의 산도가 증가했기 때문이다. 또한 30wt%를 함유한 복합 전해질막은 100℃에서는 최대 0.126 S/cm의 수소 이온 전도도를 나타내었다 PWA가 첨가됨에 따라 복합 전해질막의 열적특성 또한 증가하였다.

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.

우라늄 변환시설 가동 중 발생하여 라군(lagoon)에 저장중인 방사성 슬러지 폐기물에 대한 처리는 시설 해체과정에서 매우 중요한 업무 중 하나이다. 슬러지 구성성분 중 다량을 차지하는 질산암모늄의 폭발 위험성 등으로 인해 미생물을 이용한 질산염의 분해는 질산염을 안정적으로 처리할 수 있는 효과적인 방법이라 할 수 있다. 본 연구에서는 라군 슬러지의 약 60 wt%를 차지하는 질산염을 혐기성 균주의 하나인 Pseudomonas halodenidificans를 이용하여 탈질하기위한 공정 변수에 대한 영향을 평가하였다. 온도, 질산염 농도, 전자공여체의 영향, C/N 비율, 초기 접종하는 균주의 비율, pH등의 공정변수에 대하여 실험한 이번 결과는 향후 연속식 공정 설계를 위한 기초 자료로 사용될 것이다.

우라늄 변환시설의 라군 슬러지에 함유된 질산염의 안정적 처리를 위해 물 첨가 용해를 실시한 뒤, 여과 케이크의 안정화 특성에 대하여 알아보았다. 물 용해에 의해 대부분의 질산염은 고농도 질산염 용액으로 제거되었으므로, 여과 케이크의 열분해는 에서 하나의 단계로 수행하였다. Muffle furnace를 이용하여 에서 5시간동안 여과 케이크의 열분해를 실시한 결과 라군 1 슬러지에 포함된 U은 의 열분해와 함께 의 형태로 안정화 되었다. 라군 2 열분해 잔류물의 경우에는 열분해 시 생성된 CaO가 냉각과정에서 수분과 반응하여 로 전환되는 것을 TG/DTA 분석과 XRD 분석을 통해 확인할 수 있었지만, 처분장에서 대기중 노출이나 지하수의 침출 등에는 안정한 화합물로 알려져 있으므로, 특별한 첨가제의 첨가 없이 단순 열분해 후 처분이 가능할 것으로 판단된다.

본 논문은 수식화의 특이성 때문애 구조 최적화 문제에 거의 사용되지 않고 있는 선형 goal programrrung을 대규모 비선형 구조 최적화에 용용하는 방법올 제시한다. 이 방법은 다기준 최적화의 도구로 사용 되는데 그 까닭은 goal programming 이 목적합수와 제한조건둥을 정의하는데 있어서 발생하는 난점 들올 제거해 주기 때문이다. 이 방병은 비선형 goal 최적화 문제톨의 해톨 얻기 위해서 유한요소해석， 선형 goal programming 기볍 ‘ 그리고 계속적인 선형화 기법을 이용한다. 즉， 대규모 비선형 구조 최적화 문제를 비선형 goal programming 형태로 전환시키는 일반적인 수식화 방법을 제시하고， 얻어진 비선형 goal 최적화 문제 를 풀기 위한 계속적인 선형화 방법에 대해서도 논의한다. 얼계도구로서 이 방법의 유효성올 논증하기 위하여 10‘ 25 및 200트러스의 사례를 가지고 용력채한조건들의 최소무게 구조 최적화 문제에 대한 해를 모색하며 이 쓸 다른 연구결과와 비교검토한다.

To characterize the female or male transcriptome of the Pacific abalone and further increase genomic resources, we sequenced the mRNA of full-length complementary DNA (cDNA) libraries derived from pooled tissues of female and male Haliotis discus hannai by employing the Iso-Seq protocol of the PacBio RSII platform. We successfully assembled whole full-length cDNA sequences and constructed a transcriptome database that included isoform information. After clustering, a total of 15,110 and 12,145 genes that coded for proteins were identified in female and male abalones, respectively. A total of 13,057 putative orthologs were retained from each transcriptome in abalones. Overall Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analyzed in each database showed a similar composition between sexes. In addition, a total of 519 and 391 isoforms were genome-widely identified with at least two isoforms from female and male transcriptome databases. We found that the number of isoforms and their alternatively spliced patterns are variable and sex-dependent. This information represents the first significant contribution to sex-preferential genomic resources of the Pacific abalone. The availability of whole female and male transcriptome database and their isoform information will be useful to improve our understanding of molecular responses and also for the analysis of population dynamics in the Pacific abalone.

Background : Malonyl ginsenoside content of the Panax ginseng is known to account for 35% to 60% of total ginsenosides content. However, its distribution by ginseng part has not been studied. In this study, four kinds of malonyl ginsenosides were compared in Korean white ginseng part using the purified malonyl ginsenoside standards in our laboratory. Methods and Results : White ginseng was prepared by the air drying (50℃, 48h) or freeze drying (-70℃, 48h) methods form 4-year-old ginseng. Malonyl ginsenoside content in total ginsenosides were similar in air dried and freeze dried white ginseng, 58% and 62%, respectively. Therefore, malonyl ginsenoside contents in main, lateral, and fine root, and in the main root without skin and skin of main root prepared by freeze dried method were compared. Malonyl ginsenosides (m-Rb1, m-Rb2, m-Rc and m-Rd) and total ginsenosides (Rg1, Re, Rf, Rb1, Rc, Rb2, Rb3, Rd, m-Rb1, m-Rb2, m-Rc and m-Rd) were 6.75 and 14.15 mg/g in main root, 14.15 and 26.35 mg/g in lateral root, 46.95 and 84.15 mg/g in fine root. Malonyl ginsenoside contents in skin of main root was 20.08 mg/g, while its contents of the main root without skin was 2.58 mg/g. Conclusion : As a result, the parts each air drying the sample was confirmed that the ratio of the distribution of malonyl ginsenoside (main root : lateral : fine root = 18.7 : 11.1 : 16.2), and distribution ratio of main root, skin of main root, lateral, skin of lateral was found to be (12.2: 14.6 14.3: 3.7). Malonyl ginsenoside content was the highest in fine root, compared to the main or lateral root. Malonyl ginsenoside contents in skin of root was higher than those of the main root without skin. These results is expected to help establish an efficient extraction and standardization. Malonyl ginsenoside analysis of White ginseng using HPLC expects that the standardization process can be established.

Background : Platycodon grandiflorum is a perennial plant and a member of Campanulaceae family. Since ancient times, they have been using P. grandiflorum as an important medicinal plant in Korea. Platycodin D is the most abundant saponin derived from P. grandiflorum and pharmacologically active component. Cytochrome P450s (CYPs) are important enzymes in the saponin biosynthesis. CYP is, in general, the terminal oxidase enzymes and essential roles in saponin biosynthesis pathway by hydroxylation or oxidaition of triterpene skeletons. Methods and Results : We tried to identify CYP genes related to saponin biosynthesis of P. grandiflorum through RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 191 putative CYP genes. Familes of CYP716, CYP708, CYP93 and CYP51 were selected as putative saponin biosynthesis related gene families using phylogenetic relationship analysis. Conclusion : The results in this study could help to find the CYPs related to saponin biosynthesis pathway of P. grandiflorum.

Background : Platycodon grandiflorum is a perennial plant and a member of Camanulaceae family. Since ancient times, they have been using P. grandiflorum as an important medicinal plant in Korea. Platycodin D is the most abundant saponin derived from P. grandiflorum and pharmacologically active component. UDP-glycosyltransferases (UGTs) are important enzymes in the saponin biosynthesis. UGT is a glycosyltransferase and act on the final step of the secondary metabolite biosynthesis. Methods and Results : We tried to identify UGT genes related to saponin biosynthesis of P. grandiflorum through RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 137 putative UGT genes. Familes of UGT71, UGT73, and UGT74 were selected as putative saponin biosynthesis related gene families using phylogenetic relationship analysis. qPCR condition about UGT73 is preheating 94℃ 180 sec, denaturation 94℃ 60 sec, annealing 53℃ 60 sec, extension 72℃ 90 sec, final extension 72℃ 600 sec, 45 cycles repeated. Conclusion : The results in this study could help to find the UGTs related to saponin biosynthesis pathway of P. grandiflorum.

We recently reported rice promoters that are active in late stages of pollen development. However, rice promoters that allow manipulation of gene expression at earlier stages of pollen development are still very limited to date. In this study, we have chosen 10 putative microspore promoters, OsMSP1 through OsMSP10, based on publicly available transcriptomic datasets in rice (Oryza sativa L.). Sequence analysis of these promoter regions revealed some cis regulatory elements involved in pollen-specific expression. We also examined promoter activities using the promoter-GUS reporter constructs in both transgenic rice and Arabidopsis. In rice, all of the 10 promoters directed GUS signals from the microspore stage throughout the all stages of pollen development. In addition, while GUS signals from 4 promoters, OsMSP2, OsMSP7, OsMSP9 and OsMSP10, seem to be expressed preferentially during pollen development, those from other six promoters were observed in vegetative tissues such as leaves, stems, and roots of seedlings. Similarly, in Arabidopsis, all of the 10 promoters directed GUS signals during pollen development. In detail, 8 promoters, OsMSP1 ~ OsMSP8 directed GUS signals from the microspore stage, whereas 2 promoters, OsMSP9 and OsMSP10, exhibited GUS signals from tricellular stage. Furthermore, seven promoters, except for OsMSP1, OsMSP2 and OsMSP10, showed GUS signals in shoot apical region or root tissues of seedlings. Furthermore, we verified microspore activity of four promoters, OsMSP1, OsMSP2, OsMSP3 and OsMSP6, by complementation analysis of the sidecar pollen (scp) mutant which displays microspore-specific defects. Currently, further analyses are underway for GUS expression of T2 generation in transgenic rice and scp complementation with remaining promoters.

Rice is a staple food crop for more than half of the world population. Severe losses of rice production was caused by various environmental conditions such as cold, heat and flooding annually. Rice is a highly sensitive to low temperature below 15-20 ℃ because of originating from tropical or subtropical climates. Especially, seedling of rice is easily damaged to low temperature and result in seedling yellowing, growth retardation, reduced tillering and yield losses at last. We used a recombinant inbreeding lines (RIL) population of 384 individuals derived from a cross between Hanareum 2, a highly cold sensitive variety and Unkwang, a cold tolerant variety for molecular mapping of QTLs related to cold tolerance. Seedling discoloration of each lines and parents caused by cold response were investigated in field condition after transplanting. And leaf samples of RIL population were collected for evaluation of chlorophyll content using 80% acetone extraction. The seedling of each lines and parents was subjected to low temperate by 5~13 ℃ during 14 days. The cold recovery score (CRS) of RILs was recorded after 4 days recovery period according to standard evaluation system (SES, IRRI). Total of eight QTLs were detected on chromosome 1, 7, 8, 10, 11 and 12 using cold tolerance traits, chlorophyll content, seedling discoloration and cold recovery score in 384 RILs. The qCRS12, which detected on chromosome 12 between two flanking markers id12002113, id12002563 (1.1 Mbp) showed 25 LOD score with 26% of phenotypic variation of cold recovery score in RILs population. The positive allele contributing to cold tolerance came from the cold tolerant parent Unkwang. The result may provide useful information for a marker-assisted breeding program to improve cold tolerant in rice.

Tissue-specific promoters are a very useful tool for manipulating gene expression in a target tissue or organ; however, their range of applications in other plant species has not been determined, to date. In this study, we identified two late pollen-specific rice promoters (ProOsLPS10 and ProOsLPS11) via meta-anatomical expression analysis. We then investigated the expression of both promoters in transgenic rice (a homologous system) and Arabidopsis (a heterologous system) using ProOsLPS10 or ProOsLPS11::GFP-GUS constructs. As predicted by microarray data, both promoters triggered strong GUS expression during the late stages of pollen development in rice, with no GUS signals detected in the examined microspores and sporophytic tissues. Interestingly, these promoters exhibited different GUS expression patterns in Arabidopsis. While in Arabidopsis, the OsLPS10 promoter conferred GUS expression at the uni- and bi-cellular macrospore stages, as well as at the shoot apical region during the seedling stage, the OsLPS11 promoter was not active in the pollen at any stage, or in the examined sporophytic tissues. Furthermore, by performing a complementation analysis using a sidecar pollen (scp) mutant that displays developmental defects at the microspore stage, we found evidence that OsLPS10, which can be an applied promoter expressed in Arabidopsis, is useful for directing gene expression in the early stages of pollen development. Our results indicate that the OsLPS10 and OsLPS11 promoters can drive the expression of target genes during the late stages of pollen development in rice, but not in Arabidopsis. Our results also emphasize the necessity of confirming the applicability of an established promoter to heterologous systems.

OsLPS is pollen specific gene that express at late stage of pollen development in rice. Based on microarray database, promoter region of two genes Os03g0106900 and Os03g0106500 were identified. The sequence of 2287bp and 2468bp upstream region of these genes were amplified and designated as OsLPS10 and OsLPS11. These promoters were fused with GUS-GFP reporter gene in a destination vector, pKGWFS7 and introduced into rice (Dongjin cultivar) and Arabidopsis (Col-0). The results of GUS assay showed different pattern of gene expression in pollen of rice and Arabidopsis. In Arabidopsis, the OsLPS10 gene strongly activated in young anther and not expressed in mature pollen. Pollen development analysis revealed GUS expression was detected at unicellular stage and strongest at the bicellular pollen developmental stage. No GUS signal was recorded in mature pollen. In case of OsLPS11, no GUS signal was detected in during pollen development of inflorescent. By contrast, in rice, the GUS expression pattern of OsLPS10 and OsLPS11 exhibited similar. GUS expression was first detectable in the anthers of spikelets at the bicellular stage and intensity increased in tricellular and mature pollen. The GUS signal was not detected in the anthers in unicellular microspores in both genes, OsLPS10 and OsLPS11. The results suggested that these genes were different activity in heterologous plant system, monocot and dicot. Complementation analysis and Cis-regulatory elements will be examined to illuminate the characteristic of these genes

Based on the results of microarray analysis we selected ten candidate genes that express in pollen at the early pollen developmental stage. By PCR amplification, the promoter region of these genes were amplified from rice genomic DNA (Nipponbare) and cloned into the destination pKGWFS7 vector via an entry vector, pDONR201. The characteristic of promoters were evaluated in Arabidopsis thaliana (Col-0) through GUS expression analysis. Fifty T2 plants respectively from each promoter were tested. Whole inflorescence of individual plant was stained with 1mM X-Gluc solution to observe tissue-specific GUS expression patterns. The results showed that all 10 promoters activated in pollen tissues. Among them six promoters expressed at the early developmental stage (unicellular) of pollen and the others expressed at both early (unicellular) and late pollen developmental stage (mature pollen). The results indicated that these promoters would be potential applicable for the studies of pollen function. Currently, we are performing these promoters analysis in rice transgenic plants as well as molecular characterization.