• Title/Summary/Keyword: strain analysis

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Strain Improvement of the Genus Pleurotus by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리버섯속(屬)의 품종개발(品種開發))

  • Yoo, Young-Bok;You, Chang-Hyun;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.21 no.3
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    • pp.200-211
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    • 1993
  • Somatic hybrids of Pleurotus florida ASI 2016 and Pleurotus ostreatus ASI 2018 were obtained by protoplast fusion. The 40 fusants($P1{\sim}P40$) was examined for the yield on fermented and pasteurized rice straw in a tray. The carpophore yield of them were showed as the range of $27.0{\sim}155.2$, based on parental values of 100(ASI 2018), The pilei of fusants between orange white colored P. florida and dark grey colored P. ostreatus had mixed colors in the young stage. Other breeding programmes were performed to improve new varieties with high yield and good quality. A new oyster mushroom variety, Wonhyeongneutaribeosus(P72), was developed at the Agricultural Sciences Institute, Rural Development Administration in 1990. This P.florida-ostreatus-ostreatus hybrid P72 was selected from 38 protoplast fusion products($P41{\sim}P78$) between P.florida-ostreatus recombinant P5-M 43-arg rib and P. ostreatus ASI 2-13-0 2001-19-pro orn. The yield indexes of 38 hybrids ranged $40.5{\sim}152.7$ compared with the parental values of 100(ASI 2001). Hybrid P72 was characterized by the large fruiting bundle of semispherical shape with long stipe and by the small and circular pileus, resulting in lower harvesting cost. A significant increase in carpophore production was observed in somatic hybrids of protoplasts due to heterosis. A comparision of hybrid with parents P72 was made using isozyme analysis. The esterase banding patterns could be characterized by new bands in the hybrids. Seven fusion products of four crosses between P.florida ASI 2016 and P. ostreatus ASI 2018 were analysed with respect to the distribution of progenies and segregation of gene markers by random basidiospore analysis. Segregation of alleles should yield progeny of four genotypes in a Mendelian ratio of 1 : 1 : 1 : 1 for prototrophs, auxotrophs of one parental type, auxotrophs of the other parental type, and auxotrophic recombinants, respectively. However, five fusants of them did not detect one parental, P.ostreatus, type. Basidiospores could yield progeny of 16 genotypes in the cross of one of the recombinant P5-M43-arg $rib{\times}P. ostreatus$ ASI 2-13-pro orn but the segregants of three fusants were not detected clearly. The allele ratio of loci could be expected 1 : 1 : 1 : 1 for arg, rib, pro and orn. The ratio, however, would be changed to 4 : 1 : 1 : 1 with increasing proportion of argo In almost all the fusants, prototrophic recombinants were recovered in large numbers against auxotrophic markers. Parental genotypes were recovered with the recombinant progeny amounting to $38.68{\sim}99.56%$. The analysis provides proof of heterokaryosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis.

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Studies on Host-Virus Interaction of Poxviruses 1. Cytochemical, Autoradiographic and Immunocytological Analysis in Cowpox Virus-FL Cell System (Poxvirus 감염(感染)에 있어서의 Virus-숙주세포(宿主細胞)의 상호관계(相互關係) 1. Cowpox Virus-FL 세포계(細胞系)의 세포화학적(細胞化學的) Autoradiography 및 세포면역학적해석(細胞免疫學的解析))

  • Kim, Uh Ho
    • Korean Journal of Veterinary Research
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    • v.15 no.1
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    • pp.57-67
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    • 1975
  • The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

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Assessment of Carsington Dam Failure by Slope Stability and Dam Behavior Analyses (사면안정 해석과 댐 거동분석을 통한 Carsington Dam 파괴의 고찰)

  • 송정락;김성인
    • Proceedings of the Korean Geotechical Society Conference
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    • 1991.10a
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    • pp.87-102
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    • 1991
  • It has been reported that the failure of Carsington Dam in Eng1and occured due to the existence of a thin yellow clay layer which was not identified during the design work, and due to pre-existing shears of the clay layer. The slope stability analyses during the design work, which utilized traditional circular arc type failure method and neglected the existence of the clay layer, showed a safety factor of 1.4. However, the post-failure analyses which utilized translational failure mode considering the clay layer and the pre-existing shear deformation revealed the reduction of safety factor to unity. The post-failure analysis assumed 10。 inclination of the horizontal forces onto each slice based on the results of finite element analyses. In this paper, Bishop's simplified method, Janbu method, and Morgenstern-Price method were used for the comparison of both circular and translational failure analysis methods. The effects of the pre-existing shears and subsquent movement were also considered by varying the soil strength parameters and the pore pressure ratio according to the given soi1 parameters. The results showed factor of safefy 1.387 by Bishop's simplified method(STABL) which assumed circular arc failure surface and disregarding yellow clay layer and pre-failure material properties. Also the results showed factor of safety 1.093 by Janbu method(STABL) and 0.969 by Morgenstern-Price method(MALE) which assumed wedge failure surface and considerd yellow clay layer using post failure material properties. In addition, dam behavior was simulated by Cam-Clay model FEM program. The effects of pore pressure changes with loading and consolidation, and strength reduction near or at failure were also considered based on properly assumed stress-strain relationship and pore pressure characteristics. The results showed that the failure was initiated at the yellow clay layer and propagated through other zones by showing that stress and displacement were concentrated at the yel1ow clay layer.

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A Descriptive Statistical Analysis of the Hospitalized Patients with Low Back Pain in Departments of Korean Rehabilitation Medicine of Korean Medicine Hospitals (한국의 5개 한의과대학 부속한방병원 재활의학과의 요통 입원 환자에 대한 후향적 기술통계분석 - 입원 기간, 상병명, 치료 방법을 중심으로 -)

  • Maeng, Tae-Ho;Kim, Jongyeon;Yi, Woon-Sup;Chung, Won-Seok;Ko, Youn-Seok;Lee, Jung-Han;Shin, Byung-Cheul;Cha, Yun-Yeop;Go, Ho-Yeon;Sun, Seong-Ho;Jeon, Chan-Yong;Jang, Bo-Hyoung;Song, Yun-Kyung;Ko, Seong-Gyu
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.4
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    • pp.213-223
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    • 2013
  • Objectives Low back pain (LBP) is one of the most common reason for people in Korea to visit Korean medical institutions. To assess actual amounts of use in the treatment of LBP in Korean medicine and to provide objective base line data for policy decision making, research regarding the current state of LBP patients' treatment in Korean medical institutions are in need. Methods The current study was designed as a retrospective chart review to investigate descriptive characteristics of LBP patients. The clinical records of 304 patients who were hospitalized for the treatment of LBP in Korean rehabilitation medicine inpatient clinics of five different Korean medicine hospitals were analyzed. The percentage of patient characteristics such as sex, age, average duration of admission, insurance type, diagnosed LBP related disease code, and rates of interventions applied were assessed. Results 1. The female sex was significantly predominant among patients with LBP : 105 patients (34.5%) were male and 199 patients (65.5%) were female. Percentage of the patients' age appeared as followed : 76 people (25.0%) were in their 50s, 64 people (21.1%) were in their 40s, 51 people (16.8%) were in their 30s, 37 people (12.2%) were in their 60s, and 33 people (10.9%) were in their 70s. Approximately half of the total LBP patients investigated were older than 50. 2. The average duration of admission was 16.2 days. Approximately one third (30.3%) of the patients were hospitalized for 8 to 14 days. 3. Female patients tended to stay admitted in hospitals slightly longer than male patients. Elderly (age 60~79) patients stayed in hospitals longer (17.8 days) compared to younger (age 20~39) patients (13.5 days). 4. More than half of the patients (171 cases, 56.3%) had their hospital bills covered with automobile insurance. 40.1% (122 cases) of the patients had medical insurance to cover their hospital bills. The average duration of admission of patients who had automobile insurance was 14.2 days, while that of the patients who had medical insurance was 18.4 days. 5. "Sprain and strain of the lumbar spine and pelvis" was the most commonly used (195 cases, 64.1%) disease code in patients with LBP. Patients diagnosed as "lumbar and other intervertebral disc disorders with radiculopathy" required the longest admission duration (22.1 days). 6. Herbal medication was applied to all of the patients during admission. Acupuncture was applied to all of the patients except one case diagnosed as spinal stenosis. Physical therapy, cupping therapy, moxibustion therapy, chuna therapy, and pharmacopuncture therapy were applied to 94.7, 92.8, 85.2, 83.9, and 49.7% of the patients, respectively. 7. There were certain differences among Korean medicine hospitals in terms of the LBP patients' duration of admission, type of insurance, frequency of the disease code use, type of intervention applied. Conclusions It is thought that the current study can be used as reference data in assessing the current state of LBP treatment in Korean rehabilitation medicine and a basis for future research. Provided improvements of certain limitations of the current study in future researches, such data would act as better base line data in policy decision making.

Developmental Genetic Analysis of Avian Primordial Germ Cells and the Application to Poultry Biotechnology

  • Kagami, H.
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.135-142
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    • 2001
  • A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

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Potential of Antifungal Lactic Acid Bacteria Isolated from Kimchi as Cheese Starters (김치 분리 항진균 유산균의 치즈 스타터로서 이용 가능성)

  • Oh, Hyun Hee;Huh, Chang Ki;Choi, Ha Nuel;Yang, Hee Sun;Bae, In Hyu;Lee, Jai Sung;Jeong, Yong Seob;Lee, Nam Keun;Jung, Hoo Kil
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.133-141
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    • 2013
  • This study was performed to identify the cheese starter potential of antifungal lactic acid bacteria isolated from Kimchi. Eight fungi were isolated from cheese or the cheese ripening room, and identified as Penicillium and Cladosporium by ITS-5.8S rDNA analysis. Twenty-two lactic acid bacteria species with antifungal activity were isolated from Kimchi, and identified as Lactobacillus and Pediococcus by 16S rRNA sequence analysis. Six lactic acid bacteria species were selected (L. sakei subsp. ALJ011, L. sakei subsp. ALI033, L. sakei subsp. ALGy039, P. pentosaceus ALJ015, P. pentosaceus ALJ024, and P. pentosaceus ALJ026) based on higher antifungal activity from the initial 22 species. Out of the six identified species, L. sakei subsp. ALI033 had the highest antifungal activity. For growth of the six lactic acid bacteria, optimal temperature and pH were $30{\sim}37^{\circ}C$ and 7.0, respectively. Proteolytic activities of the six lactic acid bacteria were almost as strong as the commercial strain Str. thermophilus Body-1. Coagulative activities of L. sakei subsp. ALI033, P. pentosaceus ALJ015, and P. pentosaceus ALJ024 were higher than those of L. sakei subsp. ALJ011, L. sakei subsp. ALGy039, and P. pentosaceus ALJ026. The acid resistance of L. sakei subsp. was higher than that of P. pentosaceus. The major organic acid component of the lactic acid bacteria culture medium was lactic acid.

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Screening and Identification of a Cesium-tolerant Strain of Bacteria for Cesium Biosorption (환경유래의 세슘 저항성 균주 선별 및 세슘 흡착제거 연구)

  • Kim, Gi Yong;Jang, Sung-Chan;Song, Young Ho;Lee, Chang-Soo;Huh, Yun Suk;Roh, Changhyun
    • Korean Journal of Environmental Biology
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    • v.34 no.4
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    • pp.304-313
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    • 2016
  • One of the issues currently facing nuclear power plants is how to store spent nuclear waste materials which are contaminated with radionuclides such as $^{134}Cs$, $^{135}Cs$, and $^{137}Cs$. Bioremediation processes may offer a potent method of cleaning up radioactive cesium. However, there have only been limited reports on $Cs^+$ tolerant bacteria. In this study, we report the isolation and identification of $Cs^+$ tolerant bacteria in environmental soil and sediment. The resistant $Cs^+$ isolates were screened from enrichment cultures in R2A medium supplemented with 100 mM CsCl for 72 h, followed by microbial community analysis based on sequencing analysis from 16S rRNA gene clone libraries(NCBI's BlastN). The dominant Bacillus anthracis Roh-1 and B. cereus Roh-2 were successfully isolated from the cesium enrichment culture. Importantly, B. cereus Roh-2 is resistant to 30% more $Cs^+$ than is B. anthracis Roh-1 when treated with 50 mM CsCl. Growth experiments clearly demonstrated that the isolate had a higher tolerance to $Cs^+$. In addition, we investigated the adsorption of $0.2mg\;L^{-1}$ $Cs^+$ using B. anthracis Roh-1. The maximum $Cs^+$ biosorption capacity of B. anthracis Roh-1 was $2.01mg\;g^{-1}$ at pH 10. Thus, we show that $Cs^+$ tolerant bacterial isolates could be used for bioremediation of contaminated environments.

Cloning and Transcription Analysis of Sporulation Gene (spo5) in Schizosaccharomyces pombe (Schizosaccharomyces bombe 포자형성 유전자(spo5)의 Cloning 및 전사조절)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.112-118
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    • 2002
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by matting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB(spo5)1. We futher analyzed six recombinant plasmids, pDB(spo5)2, pDB(spo5)3, pDB(spo5)4, pDB(spo5)5, pDB (spo5)6, pDB(spo5)7 and found each of these plasmids is able to rescue the spo5-2, spo5-3, spo5-4, spo5-5, spo5-6, spo5-7 mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB(spo5)1, and pDB(spu5)Rl contained the spo5 gene. Transcripts of spo5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 2.5kb were detected with 5kb Hind Ⅲ fragment containing a part of the spo5 gene as a probe. The small mRNA(2.5kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2kb) was produced constitutively. Appearance of a 2.5kb spo5-mRNA depends upon the function of the meil, mei2 and mei3 genes.

Biological Control of Anthracnose (Colletotrichum gloeosporioides) in Red Pepper by Bacillus sp. CS-52 (Bacillus sp. CS-52를 이용한 고추 탄저병 (Colletotrichum gloeosporioides) 방제 특성)

  • Kwon, Joung-Ja;Lee, Jung-Bok;Kim, Beam-Soo;Lee, Eun-Ho;Kang, Kyeong-Muk;Shim, Jang-Sub;Joo, Woo-Hong;Jeon, Chun-Pyo;Kwon, Gi-Seok
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.201-209
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    • 2014
  • This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

Optimal Configuration of the Truss Structures by Using Decomposition Method of Three-Phases (3단계(段階) 분할기법(分割技法)에 의한 평면(平面)트러스 구조물(構造物)의 형상(形狀) 최적화(最適化)에 관한 연구(硏究))

  • Lee, Gyu Won;Song, Gi Beom
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.12 no.3
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    • pp.39-55
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    • 1992
  • In this research, a Three Level Decomposition technique has been developed for configuration design optimization of truss structures. In the first level, as design variables, behavior variables are used and the strain energy has been treated as the cost function to be maximized so that the truss structure can absorb maximum energy. For design constraint of the optimal design problem, allowable stress, buckling stress, and displacement under multi-loading conditions are considered. In the second level, design problem is formulated using the cross-sectional area as the design variable and the weight of the truss structure as the cost function. As for the design constraint, the equilibrium equation with the optimal displacement obtained in the first level is used. In the third level, the nodal point coordinates of the truss structure are used as coordinating variable and the weight has been taken as the cost function. An advantage of the Three Level Decomposition technique is that the first and second level design problems are simple because they are linear programming problems. Moreover, the method is efficient because it is not necessary to carry out time consuming structural analysis and techniques for sensitivity analysis during the design optimization process. By treating the nodal point coordinates as design variables, the third level becomes unconstrained optimal design problems which is easier to solve. Moreover, by using different convergence criteria at each level of design problem, improved convergence can be obtained. The proposed technique has been tested using four different truss structures to yield almost identical optimum designs in the literature with efficient convergence rate regardless of constraint types and configuration of truss structures.

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