• 한국균학회소식:학술대회논문집
• /
• 한국균학회 2014년도 춘계학술대회 및 임시총회
• /
• pp.27-28
• /
• 2014

Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

• Journal of Microbiology and Biotechnology
• /
• 제20권1호
• /
• pp.169-178
• /
• 2010

The microbial community structure in Thailand soils contaminated with low and high levels of arsenic was determined by denaturing gradient gel electrophoresis. Band pattern analysis indicated that the bacterial community was not significantly different in the two soils. Phylogenetic analysis obtained by excising and sequencing six bands indicated that the soils were dominated by Arthrobacter koreensis and $\beta$-Proteobacteria. Two hundred and sixty-two bacterial isolates were obtained from arsenic-contaminated soils. The majority of the As-resistant isolates were Gramnegative bacteria. MIC studies indicated that all of the tested bacteria had greater resistance to arsenate than arsenite. Some strains were capable of growing in medium containing up to 1,500 mg/l arsenite and arsenate. Correlations analysis of resistance patterns of arsenite resistance indicated that the isolated bacteria could be categorized into 13 groups, with a maximum similarity value of 100%. All strains were also evaluated for resistance to eight antibiotics. The antibiotic resistance patterns divided the strains into 100 unique groups, indicating that the strains were very diverse. Isolates from each antibiotic resistance group were characterized in more detail by using the repetitive extragenic palindromic-PCR (rep-PCR) DNA fingerprinting technique with ERIC primers. The PCR products were analyzed by agarose gel electrophoresis. The genetic relatedness of 100 bacterial fingerprints, determined by using the Pearson product-moment similarity coefficient, showed that the isolates could be divided into four clusters, with similarity values ranging from 5-99%. Although many isolates were genetically diverse, others were clonal in nature. Additionally, the arsenic-resistant isolates were examined for the presence of arsenic resistance (ars) genes by using PCR, and 30% of the isolates were found to carry an arsenate reductase encoded by the arsC gene.

• Journal of Plant Biotechnology
• /
• 제36권2호
• /
• pp.130-136
• /
• 2009

Rice (Oryza sativa) is the most important staple crop in Korea. With its small genome size of 389Mb, rice is a model plant for genome research. We analyzed expressed sequence tag (EST) clones from immature seeds of rice (cv. Ilpum) at 20 days after heading. The 25,668 EST clones were clustered by using SeqMan program and 7,509 clones were selected as unique clones. We compared the 7,509 unique genes with KOME database including the 32,127 FL-cDNA in rice. Finally, 4,990 clones were homologous and 2,519 clones non-homologous to FL-cDNA clones. In addition, we mapped the 7,509 cDNA clones by using TIGR rice pseudomolecule version 5. Ultimately, 7,347 clones were matched to be significant clones related to the TIGR rice pseudomolecules, but 162 clones were unmapped. For the clustering of orthologous group genes, we further analyzed the 7,509 EST clones from immature seeds using NCBI clusters of orthologous groups database. Among the clones, 4,968 clones were categorized into information storage and processing, cellular processes and signaling, metabolism and poorly characterized genes, proportioning 799 (14.89%), 1,536 (28.3%), 1,148 (21.2%) and 1,936 (35.7%) clones to the previous four categories, respectively.

• 공업화학
• /
• 제7권3호
• /
• pp.464-472
• /
• 1996

이온화 클러스터 빔 증착에 의한 박막 형성 기구를 연구하였다. 가속전압, 기판온도, 표면이동 활성화에너지, 임계핵 크기 등이 박막의 결정입자크기 및 표면 평활도에 미치는 영향을 조사하기 위하여 Monte-Carlo방법에 근거한 전산 모사 프로그램을 개발하였다. 클러스터의 운동에너지가 박막형성에 미치는 영향을 관찰함으로써 높은 가속전압이 입자들의 표면이동에너지를 증가시키며 단결정막의 생성을 용이하게 함을 알 수 있었다. 기판온도가 증가함에 따라 입자의 운동에너지 소산(dissipation)속도는 느려지고 따라서 결정입자의 크기가 커졌다. 이러한 효과는 임계핵 크기가 클수록 두드러졌다. 표면이동 활성화에너지는 입자와 기판간의 상호작용력과 운동에너지 소산속도에 영향을 미침이 발견되었다. 가속전압, 기판온도, 표면이동 활성화에너지는 박막의 평활도에 복합적인 영향을 미치는 것을 알 수 있었다.

• 한국국제농업개발학회지
• /
• 제31권4호
• /
• pp.378-383
• /
• 2019

본 연구는 FT-IR 스펙트럼 데이터를 기반으로 다변량통계분석을 이용하여 생육 온도변화에 따른 파파야(Carica papaya L.)의 대사체 수준 식별을 통해 기후 변화에 대응하여 작물의 육종 연구의 기초자료로 활용하고자 한다. 1. FT-IR 스펙트럼 데이터로부터 PCA(principal component analysis), PLS-DA(partial least square discriminant analysis) 그리고 HCA(hierarchical clustering analysis) 분석을 실시하였다. 2. 파파야 품종은 1700-1500, 1500-1300, 1100-950 cm^-1부위에서 대사체의 양적, 질적 패턴 변화가 FT-IR 스펙트럼상에서 나타났다. FT-IR 스펙트럼의 1700-1500 cm^-1부위는 주로 Amide I 과 II을 포함하는 아미노산 및 단백질계열의 화합물들의 질적, 양적 정보를 나타내고, 1500-1300 cm^-1부위는 phosphodiester group을 포함한 핵산 및 인지질의 정보가 반영이 되고, 1100-950 cm^-1부위는 단당류나 복합 다당류를 포함하는 carbohydrates 계열의 화합물들이 질적, 양적 정보가 반영되는 부위이다. 3. PCA score plot 상측으로부터 +0℃(A)에서 +4℃(C)로 변화하는 것을 볼 수 있다. (A) 그룹은 주로 현재 기온에서 재배되는 파파야가 분포되면서 그룹을 형성하고 있고, (B) 그룹은 평년 기온에서 +2℃ 증가한 것을 가정하여 재배된 파파야가 그룹을 형성하였다. 또한, (C) 그룹은 (B) 그룹에서 +2℃, 평년 기온에서 +4℃ 증가한 것을 가정하여 재배된 파파야가 그룹을 형성하였다. 4. PLS-DA 분석의 경우 PCA 분석보다 생육온도에 따른 그룹 간 식별이 뚜렷하게 나타났다. 5. 본 연구에서 확립된 파파야 생육온도에 따른 대사체 수준 식별 기술은 파파야의 품종, 계통의 신속한 선발 수단으로 활용이 가능할 것으로 기대되며 육종을 통한 신품종개발 가속화에 기여할 수 있을 것으로 예상된다.

• 한국균학회소식:학술대회논문집
• /
• 한국균학회 2015년도 춘계학술대회 및 임시총회
• /
• pp.45-46
• /
• 2015

Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

• Journal of Plant Biotechnology
• /
• 제36권3호
• /
• pp.281-288
• /
• 2009

Seed germination is the important stage to express many genes for regulation of energy metabolism, starch degradation and cell division from seed dormancy state. For the functional analysis of seed germination mechanisms, we were analyzed the rice cDNA clones (Oryzasativa cultivar Ilpum) obtained from seed imbibition during 48 hours. Total number of 18,101 Expressed Sequence Tags (ESTs) were clustered using SeqMan program. Among them, 8,836 clones were identified as unique clones. We identified the chitinase gene specifically expressed in seed germination and amylase gene involved to starch degradation from the full length cDNA analysis, and several genes were registered to NCBI GeneBank. To analyzed the commonly expressed genes between inmature seed and germinated seed, 25,66 inmature ESTs and 18,101 germinated ESTs were clustered using SeqMan program and identified 2,514 clones as commonly expressed unigene. Among them, alpha-glubulin and alcohol dehydrogenase I were supposed to LEA genes only expressed in the immature and germinated seed stages. For the clustering of orthologous group genes, we further analyzed the 8,836 EST clones from germinating seeds using NCBI clusters of orthologous groups database. Among the clones, 5,076 clones were categorized into information storage and processing, cellular processes and signaling, metabolism and poorly characterized genes, proportioning 783 (14.29%), 1,484 (27%), 1,363 (24.8%) and 1,869 (34%) clones to the previous four categories, respectively.

• 한국재료학회:학술대회논문집
• /
• 한국재료학회 2012년도 춘계학술발표대회
• /
• pp.68.1-68.1
• /
• 2012

The best part of graphene is - charge-carriers in it are mass less particles which move in near relativistic speeds. Comparing to other materials, electrons in graphene travel much faster - at speeds of $10^8cm/s$. A graphene sheet is pure enough to ensure that electrons can travel a fair distance before colliding. Electronic devices few nanometers long that would be able to transmit charge at breath taking speeds for a fraction of power compared to present day CMOS transistors. Many researches try to check a possibility to make it a perfect replacement for silicon based devices. Graphene has shown high potential to be used as interconnects in the field of high frequency electrical devices. With all those advantages of graphene, we demonstrate characteristics of electrical and optical properties of graphene such as the effect of graphene geometry on the microwave properties using the measurements of S-parameter in range of 500 MHz - 40 GHz at room temperature condition. We confirm that impedance and resistance decrease with increasing the number of graphene layer and w/L ratio. This result shows proper geometry of graphene to be used as high frequency interconnects. This study also presents the optical properties of graphene oxide (GO), which were deposited in different substrate, or influenced by oxygen plasma, were confirmed using different characterization techniques. 4-6 layers of the polycrystalline GO layers, which were confirmed by High resolution transmission electron microscopy (HRTEM) and electron diffraction analysis, were shown short range order of crystallization by the substrate as well as interlayer effect with an increase in interplanar spacing, which can be attributed to the presence of oxygen functional groups on its layers. X-ray photoelectron Spectroscopy (XPS) and Raman spectroscopy confirms the presence of the $sp^2$ and $sp^3$ hybridization due to the disordered crystal structures of the carbon atoms results from oxidation, and Fourier Transform Infrared spectroscopy (FTIR) and XPS analysis shows the changes in oxygen functional groups with nature of substrate. Moreover, the photoluminescent (PL) peak emission wavelength varies with substrate and the broad energy level distribution produces excitation dependent PL emission in a broad wavelength ranging from 400 to 650 nm. The structural and optical properties of oxygen plasma treated GO films for possible optoelectronic applications were also investigated using various characterization techniques. HRTEM and electron diffraction analysis confirmed that the oxygen plasma treatment results short range order crystallization in GO films with an increase in interplanar spacing, which can be attributed to the presence of oxygen functional groups. In addition, Electron energy loss spectroscopy (EELS) and Raman spectroscopy confirms the presence of the $sp^2$ and $sp^3$ hybridization due to the disordered crystal structures of the carbon atoms results from oxidation and XPS analysis shows that epoxy pairs convert to more stable C=O and O-C=O groups with oxygen plasma treatment. The broad energy level distribution resulting from the broad size distribution of the $sp^2$ clusters produces excitation dependent PL emission in a broad wavelength range from 400 to 650 nm. Our results suggest that substrate influenced, or oxygen treatment GO has higher potential for future optoelectronic devices by its various optical properties and visible PL emission.

• 한국산학기술학회논문지
• /
• 제7권2호
• /
• pp.120-125
• /
• 2006

산업적으로 경제적인 급속 응고법과 $450^{\circ}C-1hr$ 대기 어닐링을 이용하여 ${\pm}5000Oe$ 이상의 고자계에서 5% 이상의 거대 자기효과를 보이는 두께 $20{\mu}m$의 granular CuCo 합금 리본을 제작하였다. Co의 함량에 따른 최대 자기저항 효과를 확인하기 위해서 Co의 함량을 $5{\sim}30at%$ 범위에서 변화시키며 아래의 자기저항효과와 포화자화의 자기적 특성과 미세구조를 확인하였다. 최대자기저항 효과는 Cu-10at%Co에서 1.2T에서 5.2%의 GMR ratio를 얻었으며, 0.5T에서는 $Cu-8{\sim}14at%Co$에서 공업적으로 활용이 가능한 3% 이상의 MR ratio를 확보할 수 있었다. 5% 이하의 Co 조성범위 리본에서는 초상자성(superparamagnetic)때문에, 20%이상의 Co 조성의 리본에서는 포화자화는 커지지만 Co 클러스터의 과도 성장으로 MR 효과가 급격히 감소하였다. 대기 분위기의 급속 응고와 열처리에 의한 표면 산화막은 자기저항효과에 큰 영향을 주지 못하였다. 따라서 고자계 센서로 사용되는 CuCo 그래뉼라 합금 리본은 $8{\sim}14wt%Co$의 조성 범위에서 공업적 목적에 유리한 거대 자기저항을 갖는 것이 가능하였다.

• Journal of Plant Biotechnology
• /
• 제44권4호
• /
• pp.416-430
• /
• 2017

The kenaf plant is used widely as food and in traditional folk medicine. This study evaluated the morphological characteristics, functional compounds, and genetic diversity of 32 kenaf cultivars from a worldwide collection. We found significant differences in the functional compounds of leaves from all cultivars, including differences in levels of chlorogenic acid isomer (CAI), chlorogenic acid (CA), kaempferol glucosyl rhamnoside isomer (KGRI), kaempferol rhamnosyl xyloside (KRX), kaemperitrin (KAPT) and total phenols (TPC). The highest TPC, KAPT, CA, and KRX contents were observed in the C22 cultivars. A significant correlation was observed between flowering time and DM yield, seed yield, and four phenolic compounds (KGRI, KRX, CAI, and TPC) (P < 0.01). To assess genetic diversity, we used 80 simple sequence repeats (SSR) primer sets and identified 225 polymorphic loci in the kenaf cultivars. The polymorphism information content and genetic diversity values ranged from 0.11 to 0.79 and 12 to 0.83, with average values of 0.39 and 0.43, respectively. The cluster analysis of the SSR markers showed that the kenaf genotypes could be clearly divided into three clusters based on flowering time. Correlations analysis was conducted for the 80 SSR markers; morphological, chemical and growth traits were found for 15 marker traits (corolla, vein, petal, leaf, stem color, leaf shape, and KGRI content) with significant marker-trait correlations. These results could be used for the selection of kenaf cultivars with improved yield and functional compounds.