• Title/Summary/Keyword: mass selection

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Biosurfactant as a microbial pesticide

  • Lee, Baek-Seok;Choi, Sung-Won;Choi, Ki-Hyun;Lee, Jae-Ho;Kim, Eun-Ki
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.40-44
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    • 2003
  • Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

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Expression of Recombinant Erythropoietin Gene in Transgenic Tobacco Plant (형질전환 담배 식물체에서 재조합 erythropoietin 유전자의 발현)

  • CHOI, Jang Won;PARK, Hee Sung
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.63-69
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    • 1997
  • Erythropoietin (EPO) is a glycoprotein that mediates the growth and differentiation of erythroid progenitors. In order to produce recombinant human erythropoietin in tobacco plant, the EPO genomic DNA (5.4 kb) was cloned into plant expression vectors, pBI$\Delta$GUS121, pBD$\Delta$GUS121 and pPEV-1, and introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated transformation. After selection on MS media containing kanamycin (Km), 10 Km-resistant plants were obtained per each construct. The correct integration of EPO genomic DNA in the genome of transgenic plant was confirmed by polymerase chain reaction (PCR). Northern blot showed that transcripts of 1.8 kb length were produced in leaves of the plants, but there was no difference of mRNA amount according to promoter number and 5'-untranslated sequence (UTS). The proteins obtained from leaves of transgenic plants were immunologically detected by Western blot using rabbit anti-human EPO polyclonal antibody. The expressed protein appeared as smaller band of apparent mass of 30 kDa as compared to the EPO protein from human urine (37 kDa), suggesting that the modification (glycosylation) system in tobacco plant might be different from that of mammalian cells.

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Growth Characteristics of Microalgae Scenedesmus obliquus by LED Light Source (LED 광원에 따른 미세조류 Scenedesmus obliquus의 성장 특성)

  • Yoo, Yong Jin;Kim, Song Yi;Lee, Geon Woo;Lee, Young Bok;Kim, Jin Woo;Kim, Ho Seob
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.11
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    • pp.70-77
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    • 2020
  • Microalgae are independent organisms that perform photosynthesis and can alter the culture environment to increase accumulation of useful substances derived from microalgae. In this study, cell growth was measured by incubation for 39 days using MBBM, Neo medium, and seven light sources, which is the main factor affecting cell growth of microalgae S. obliquus. In the case of S. oliquus, which grew in MBBM and Neo medium, cell growth was highest under fluorescent light sources and Red2 LED (R660) light sources, and cell growth was lowest under Infra Red LED (R741) light sources. The average cell growth rate was 17.7% for MBBM and 15.4% for Neo. Comparing the effects of dry cell weight of Neo medium containing nutrients on the production of aquatic plants, MBBM and dry cell weight of Neo resulted in higher cell growth than Neo medium under all LED light sources except for Blue LED (B450). This proves that MBBM is more suitable for increasing the cell growth of microalgae than Neo medium and confirms that light source selection is important in the production of useful materials through mass cultivation of microalgae in the future.

GFP expression in the microspore-derived early embryo through co-culturing with Agrobacterium (Agrobacterium 공동배양을 이용한 고추 소포자 유래 초기 배의 GFP 발현)

  • Jung, Min;In, Dong-Su;Kim, Bong-Kyu;Jang, In-Chang;Park, Eun-Joon;Kim, Moon-Za;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.109-114
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    • 2008
  • The aim of this research is to establish the conditions for Agrobacterium-mediated genetic transformation using microspore. The embryo induction from the microspore was examined under several Kanamycin concentration in media, and the induction rate decreased about 4, 8, 10 times when the Kanamycin concentration increased 10, 50, 100 mg/L, respectively. This indicates that the transformation rate would be much lower if the Kanamycin was used for selection marker. In order to apply the GFP gene as a reporter gene for Agrobacterium-mediated genetic transformation, GFP expression from the microspore-mediated embryos was observed using GFP filter under microscope. The GFP expression occurred when the microspore cultured toward the embryo development for 12, 24 and 48 days. The microspore formed a cluster by microspore division from 12 days culture and continuously became a bigger mass. We obtained a total of 8 GFP-expressing embryos suggesting that the transformation of microspore occurred. However, those young embryos were not fully developed. Further study pertinent to culture conditions is required to fulfill the Agrobacterium-mediated genetic transformation using microspore.

Effect of a Common Medium on the Growth of Nitrogen Fixer Rhizobium and Phosphate Solubilizer Bacillus megaterium (질소고정균(Rhizobium)과 인산가용화균(Bacillus megaterium)의 동시배양을 위한 배양조건 탐색)

  • Poonguzhali, Selvaraj;Thangaraju, Muthu;Ryu, Jyung-Hyun;Madhaiyan, Munusamy;Chung, Keun-Yook;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.1
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    • pp.8-14
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    • 2005
  • Mass culturing of two beneficial organisms used as biofertilizers for crops would reduce the risks in production and minimize the capital involved and this demands appropriate media that supports both organism and also selection of organisms that are not antagonistic to each other. A study was initiated to culture a nitrogen fixer (Rhizobium) and phosphate solubilizer (Bacillus megaterium) in a single medium and to study their growth patterns and shelf life in carrier. The growth of Rhizobium and Bacillus megaterium was assessed in different media and a slight modification in the traditional yeast extract mannitol media promoted the growth of both the organisms. The growth of the individual organisms in the modified medium was assessed by estimating the population at regular intervals and compared to their original medium. Maximum population of Rhizobium and phosphobacteria was at 60 hr when the phosphiobacteria inoculation of later was after 48 hr of Rhizobium inoculation. The shelf life of the individual inoculants in the inoculant containing both the organism in a sterile carrier base revealed no significant differences compared to individual organisms inoculated in a sterilized carrier. The population of both organisms in carrier based mixed inoculant remained at $10^8$ cells till 90 days.

Selection and Characteristics of Bacteriocin-Producing Microorganism to Utilize in Anti-Bacterial Rice Brain Protein Film Production (항균성 미강 단백질 필름 개발을 위한 Bacteriocin 생성균주의 선별 및 특성)

  • Kim, Eun-Joung;Kim, Kyung-Mi;Han, Hye-Kyung;Kim, Young-Ho;Kwon, Ki-Sung;Bae, Dong-Ho
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.285-290
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    • 2003
  • This study was conducted to select the bacteriocin-producing microoreanism cultivated in the rice bran culture and to characterize the produced bacteriocin for the further purpose of economical and anti-bacterial rice bran protein film. Pseudomonas putida 21025 was cultivated from rice bran and identified as a producer of a bacteriocin which showed bactericidal activity against Pseudomonas aeruginosa 9027. Bacteriocin produced by Pseudomonas putida 21025 showed a broad spectrum of activity against spoilage and soil bacteria. The activity of the bacteriocin produced by Pseudomonas putida 21025 decreased after 1 hr of staying at the temperature of $50^{\circ}C$, and with the presence of some organic solvents, except hexane and ethanol. However, the bacteriocin activity was stable throughout the pH ranges of 6-9 for 2 hrs, at the temperature lower than $50^{\circ}C$, and with the presence of ethanol for 3 hrs. The bacteriocin was partially purified by 50% ammonium sulfate precipitation followed by subsequent dialysis. Direct detection of the partially purified bacteriocin on SDS-PAGE suggested that it had an apparent molecular mass of about 21.6 kDa.

Development of a Bioassay Method Using Aluminium Foil Sheet for Screening Ovicidal Activity Against Diamondback Moth Egg, Plutella xylostella L. and Selection of Plant Extracts with High Ovicidal Activity (알루미늄호일 이용 배추좀나방(Plutella xylostella L.) 살란활성 검정법 개발 및 살란활성 식물추출물의 선발)

  • Kwon, Min;Kim, Ju-Il;Lee, Seung-Hwan;Lin, Mei-Ying
    • The Korean Journal of Pesticide Science
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    • v.15 no.2
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    • pp.194-200
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    • 2011
  • Diamondback moth (DBM, Plutella xylostella L.) is known as the most destructive pest of cruciferous crops worldwide. As most insecticides targeted to mainly larval stage, new insecticides which have hatching-inhibitory or ovicidal activity could be more efficient to control DBM. Therefore, we developed an easy and efficient method for screening ovicidal activity of DBM eggs using aluminum foil. The aluminum foil ($4{\times}12$ cm) coated with filtered juice of chinese cabbage leave (hereinafter called oviposition foil) exposed to 300 newly-emerged adults for 24 hours inside the rearing container. The oviposition foils were replaced every 4 days consecutively after mating, but it was better to discarded over then. Oviposition foil were divided into 6 to 12 pieces depending on egg mass volume. After dipping into test solutions for 10 seconds using faucet, oviposition foil pieces were placed into common petri dish, and then investigated hatchability. The effect of methanol solvent (50%) for 10 seconds dipping on the toxicity against DBM eggs was negligible. In addition, whether covering the petri dish or not should be dependent on nature of active compounds tested. With applying the new bioassay method, methanol extracts from 50 plants were tested the ovicidal activity to DBM eggs. Among them, four plant extracts; Angelica tenuissima root, Lycium chinense root, Cnidium officinale root and Polygala tenuifolia root, showed high ovicidal activity of over 90% control efficacy, against DBM eggs.

A Study on the Determination of Reference Parameter for Aircraft Impact Induced Risk Assessment of Nuclear Power Plant (원전의 항공기 충돌 리스크 평가를 위한 대표매개변수 선정 연구)

  • Shin, Sang Shup;Hahm, Daegi;Choi, In-Kil
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.27 no.5
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    • pp.437-450
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    • 2014
  • In this study, we developed a methodology to determine the reference parameter for an aircraft impact induced risk assessment of nuclear power plant (NPP) using finite element impact analysis of containment building. The target structure used to develop the method of reference parameter selection is one of the typical Korean PWR type containment buildings. We composed a three-dimensional finite element model of the containment building. The concrete damaged plasticity model was used for the concrete material model. The steels in the tendon, rebar, and liner were modeled using the piecewise-linear stress-strain curves. To evaluate the correlations between structural response and each candidate parameter, we developed Riera's aircraft impact force-time history function with respect to the variation of the loading parameters, i.e., impact velocity and mass of the remaining fuel. For each force-time history, the type of aircraft is assumed to be a Boeing 767 model. The variation ranges of the impact velocity and remaining fuel percentage are 50 to 200m/s, and 30 to 90%, respectively. Four parameters, i.e., kinetic energy, total impulse, maximum impulse, and maximum force are proposed for candidates of the reference parameter. The wellness of the correlation between the reference parameter and structural responses was formulated using the coefficient of determination ($R^2$). From the results, we found that the maximum force showed the highest $R^2$ value in most responses in the materials. The simplicity and intuitiveness of the maximum force parameter are also remarkable compared to the other candidate parameters. Therefore, it can be concluded that the maximum force is the most proper candidate for the reference parameter to assess the aircraft impact induced risk of NPPs.

A Study on the Development of Human Resources and Management System Model for Home-based Welfare Services in Pusan (재가복지서비스를 위한 인적자원개발 및 관리체계모형에 관한 연구 -부산지역을 중심으로-)

  • Ryu, Ki-Hyung;Kim, Ki-Tae;Park, Byung-Hyun;Park, Kyung-Ill;Lee, Kyung-Hee
    • Korean Journal of Social Welfare
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    • v.36
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    • pp.101-127
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    • 1998
  • The purpose of this study is to develop human resources and management system model for home-based welfare services. For the purpose of the study, both of method of literature review about home-based welfare services and previous research and survey were used in the study. The subjects of the survey were Junior and senior high school students, the employed, housewives, the disabled, and the aged. They were considered as human resources for the home-based welfare services. Based on the result of the survey and related theories, the study suggests the model for the development of volunteers and management system composed of recruitment, education and training, disposition, management and supervision, and evaluation for home-based welfare services. Assuming linking and cooperation between volunteer center and agency using volunteers which provide home-based welfare services, the model was developed. The role of volunteer center is recruitment, education and training. The agency using volunteers is responsible for education, disposition, supervision, and evaluation related to practice. In the stage of recruitment, elaborating strategy considering characteristics of group was suggested. In the campaign, use of mass media, selection of timing, equal opportunity for all group were suggested. Also outreach strategy as a effective recruitment strategy was presented. In the stage of education and training, method and content of education in each stage of primary education and re-education were presented. In the stage disposition, method and content for the right volunteer in the right place were suggested. In the stage of management and supervision, supervision by coordinator and necessity of using volunteers as para-professionals was suggested. Also the necessity of program operation for volunteer maintenance and management was discussed. In the stage of evaluation, time, main body, and method of evaluation was suggested.

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Occurrence Ecology of Ricania sp. (Hemiptera: Ricaniidae) and Selection of Environmental Friendly Agricultural Materials for Control (갈색날개매미충(신칭, Ricania sp.)의 발생생태와 친환경 방제자재 선발)

  • Choi, Duck-Soo;Kim, Do-Ik;Ko, Sug-Ju;Kang, Beom-Ryong;Lee, Kwan-Seok;Park, Jong-Dae;Choi, Kyeong-Ju
    • Korean journal of applied entomology
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    • v.51 no.2
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    • pp.141-148
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    • 2012
  • An outbreak of Ricania sp. occurred in the Kurye, Jeonnam area in 2011. This outbreak damaged many kinds of fruit trees such as Cornus, Persimmon and Chestnut. This experiment was conducted to survey the occurrence ecology of Ricania sp. such as host plants, oviposition characters, morphological characters and life cycle, as well as to select environmental friendly control agents. Ricaina sp. host plants included 51 species such as 32 xylophytes, and 19 herbaceous plants. Ricaina sp. preferred Cornus officinalis, Diospyros kaki, Castanea crenata, Eucommia ulmoides, Styrax japonicus for oviposition. Adults laid eggs on new inner twigs with 28.8 eggs per egg-mass. Egg size was 1.24 mm(length), 0.55 mm(width) in an oval shape. Nymphs molted four times. Every nymph stage had an x shape of yellow or white beeswax around the anus. Overwintered eggs of Ricania sp. hatched from the mid May to early June. Nymphal periods were from mid May to mid August and adults appeared from mid July but spawning began in mid August. Ricania sp. damaged new twigs by oviposition and retarded growth by sucking nutrients and producing a sooty mold. Sophora and natural plant extracts were effective environmentally friendly agricultural materials used to control the nymph and adult Ricania sp. Mortality was > 80%.