• Title/Summary/Keyword: Inoculation density

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Involvement of Growth-Promoting Rhizobacterium Paenibacillus polymyxa in Root Rot of Stored Korean Ginseng

  • Jeon, Yong-Ho;Chang, Sung-Pae;Hwang, In-Gyu;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.881-891
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    • 2003
  • Paenibacillus polymyxa is a plant growth-promoting rhizobacterium (PGPR) which can be used for biological control of plant diseases. Several bacterial strains were isolated from rotten roots of Korean ginseng (Panax ginseng C. A. Meyer) that were in storage. These strains were identified as P. polymyxa, based on a RAPD analysis using a P. polymyxa-specific primer, cultural and physiological characteristics, an analysis utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME), and the 16S rDNA sequence analysis. These strains were found to cause the rot in stored ginseng roots. Twenty-six P. polymyxa strains, including twenty GBR strains, were phylogenetically classified into two groups according to the ERIC and BOX-PCR analyses and 16S rDNA sequencing, and the resulting groupings systematized to the degrees of virulence of each strain in causing root rot. In particular, highly virulent GBR strains clustered together, and this group may be considered as subspecies or biovar. The virulence of the strains seemed to be related to their starch hydrolysis enzyme activity, but not their cellulase or hemicellulase activity, since strains with reduced or no starch-hydrolytic activity showed little or no virulence. Artificial inoculation of the highly virulent strain GBR-1 onto the root surfaces of Korean ginseng resulted in small brown lesions which were sunken and confined to the outer portion of the root. Ginseng root discs inoculated in vitro or two-year-old roots grown in soil drenched with the inoculum developed significant rot only when the inoculum density was $10^{6}-10^{7}$ or more colony-forming units (CFU) per ml. These results suggest that P. polymyxa might induce ginseng root rot if their population levels are high. Based on these results, it is recommended that the concentration of P. polymyxa should be monitored, when it is used as a biocontrol agent of ginseng, especially in the treatment of stored roots.

Comparative Study on the Characteristics of Microalgae as Standard Species for Marine Ecotoxicity Tests (Skeletonema sp., Dunaliella tertiolecta) (해양생태독성시험 표준생물로서 미세조류의 특성 비교 연구(Skeletonema sp., Dunaliella tertiolecta))

  • Kim, Tae Won;Moon, Chang Ho;Lee, Su Jin
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.26 no.5
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    • pp.514-522
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    • 2020
  • To understand the ecotoxicological differences between representative Skeletonema sp. and Dunaliella tertiolecta, both producers as international standard test species for marine ecotoxicity testing, we compared each standard test method, and comparatively analyzed the suitability of the species for environmental assessment and their sensitivity to various test substances. Although most of the test conditions were the same in each method, there were differences in limitation of pH changing and the initial inoculation density in the validation criteria, which is supposed to originate from the low growth rate of D. tertiolecta. In terms of suitability, both species showed consistency in test performance by repeatedly meeting the validation criteria required by the standard test methods. The salinity ranges available for testing were 20 and 10 psu for Skeletonema sp. and D. tertioelecta, respectively. Finally, regarding sensitivity, the toxicity sensitivity of Skeletonema sp. was relatively higher than that of D. tertiolecta for the reference toxicant, actual polluted water discharged (ballast water), and other chemicals. This implies that using at least two species of microalgae from different classification groups could help increase the reliability and objectivity of test results in the performance of marine ecotoxicity tests using producers.

Modelling The Population Dynamics of Laodelphax striatellus Fallén on Rice (벼에서 애멸구(Laodelphax striatellus Fallén) 개체군 밀도 변동 예측 모델 구축)

  • Kwon, Deok Ho;Jeong, In-Hong;Seo, Bo Yoon;Kim, Hey-Kyung;Park, Chang-Gyu
    • Korean journal of applied entomology
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    • v.58 no.4
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    • pp.347-354
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    • 2019
  • Temperature-dependent traits of Laodelphax striatellus, rice stripe virus vector, were investigated at 10 constant temperatures (12.5, 15.0, 17.5, 20.0, 22.5, 25.0, 27.5, 30.0, 32.5, and 35.0 ± 1℃) under a fixed photoperiod (14/10-hr light/dark cycle). Unit functions for the oviposition model were estimated and implemented into a population dynamics model using DYMEX. The longevity of L. striatellus adults decreased with increasing temperature (56.0 days at 15.0℃ and 17.7 days at 35.0℃). The highest total fecundity (515.9 eggs/female) was observed at 22.5℃, while the lowest (18.6 eggs/female) was observed at 35.0℃. Adult developmental rates, temperature-dependent fecundity, age-specific mortality rates, and age-specific cumulative oviposition rates were estimated. All unit equations described adult performances of L. striatellus accurately (r2 =0.94~0.97). After inoculating adults, the constructed model was tested under pot and field conditions using the rice-plant hopper system. The model output and observed data were similar up to 30 days after inoculation; however, there were large discrepancies between observed and estimated population density after 30 days, especially for 1st and 2nd instar nymph densities. Model estimates were one or two nymphal stages faster than was observed. Further refinement of the model created in this study could provide realistic forecasting of this important rice pest.

Large-scale Culture of Plant Cell and Tissue by Bioreactor System

  • Son, Sung-Ho;Park, Sung-Mee;Park, Seung -Yun;Kwon, Oh-Woung;Lee, Yun-Hee;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.1-7
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    • 1999
  • Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

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Menadione Sodium Bisulfite-Protected Tomato Leaves against Grey Mould via Antifungal Activity and Enhanced Plant Immunity

  • Jo, Youn Sook;Park, Hye Bin;Kim, Ji Yun;Choi, Seong Min;Lee, Da Sol;Kim, Do Hoon;Lee, Young Hee;Park, Chang-Jin;Jeun, Yong-Chull;Hong, Jeum Kyu
    • The Plant Pathology Journal
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    • v.36 no.4
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    • pp.335-345
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    • 2020
  • Tomato grey mould has been one of the destructive fungal diseases during tomato production. Ten mM of menadione sodium bisulfite (MSB) was applied to tomato plants for eco-friendly control of the grey mould. MSB-reduced tomato grey mould in the 3rd true leaves was prolonged at least 7 days prior to the fungal inoculation of two inoculum densities (2 × 104 and 2 × 105 conidia/ml) of Botrytis cinerea. Protection efficacy was significantly higher in the leaves inoculated with the lower disease pressure of conidial suspension compared to the higher one. MSB-pretreatment was not effective to arrest oxalic acid-triggered necrosis on tomato leaves. Plant cell death and hydrogen peroxide accumulation were restricted in necrotic lesions of the B. cinereainoculated leaves by the MSB-pretreatment. Decreased conidia number and germ-tube elongation of B. cinerea were found at 10 h, and mycelial growth was also impeded at 24 h on the MSB-pretreated leaves. MSB-mediated disease suppressions were found in cotyledons and different positions (1st to 5th) of true leaves inoculated with the lower conidial suspension, but only 1st to 3rd true leaves showed decreases in lesion sizes by the higher inoculum density. Increasing MSB-pretreatment times more efficiently decreased the lesion size by the higher disease pressure. MSB led to inducible expressions of defence-related genes SlPR1a, SlPR1b, SlPIN2, SlACO1, SlChi3, and SlChi9 in tomato leaves prior to B. cinerea infection. These results suggest that MSB pretreatment can be a promising alternative to chemical fungicides for environment-friendly management of tomato grey mould.

Comparative Genetic Characterization of Plasmids of Agrobacterium Species Isolated in Korea (한국산 Agrobacterium plasmid의 유전학적 성상에 관한 연구)

  • Kim, Jung-Hye;Koo, Yong-Bum;Lee, Ki-Yung;Chung, Jae-Kyu
    • Journal of Yeungnam Medical Science
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    • v.1 no.1
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    • pp.41-48
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    • 1984
  • The soil bacterium Agrobacterium tumefaciens is a plant pathogen that cause3 crown gall tumors by infecting the wounded dicotyledonous plants and subsequent integration of bacterial DNA into plant nuclear DNA. Virulent A. tumefaciens strains harbor a large Ti (tumor-inducing) plasmid that carries genes essential for tumorigenesis. In the present study, 13 strains (Malus pumila Mill; $A_{1-3}$, Populus monilifera; $W_{1-6}$, Populus tomentiglandlosa; $P_{1-3}$ and Rosa species; $R_1$) of Agrobacterium isolated in korean crown gall tumors and plasmids were observed in 6 strains ($W_2$, $W_3$, $W_6$, $P_1$, $P_3$ and $A_2$). The test for crown gall tumor formation was resulted only in ATCC15955 and $KW_2$ strains inoculated into the stem of sun flower and the development was observed for 4 and 6 weeks after inoculation. Above two Ti plasmids (pTi) were purified by cesium chloride-ethidium bromide density gradient centrifugation and digested with restriction enzyme and fragments of pTiATCC 15955 and $pTiKW_2$ observed by EcoR I ; 25&27, Hind III; 23&21, BamH I ; each 20 and Hpa I ; 12&27, and sizes of pTiATCC15955 and and $pTiKW_2$ calculated as 200 and 87 kbases. Octopine was isolated from tumor tissue ($W_{1-6}$ and $P_{1-3}$) and these strains confirmed as octopine type.

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Growth characteristics of oyster mushroom upon addition of barley flour as a substitute for cotton seed meal (면실박 대체 보리가루 첨가에 따른 느타리버섯의 생육 특성)

  • Lee, Chan-jung;Lee, Eun-ji;Park, Hye-sung;Kong, Won-sik
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.145-149
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    • 2017
  • This study was conducted to investigate the optimum additive ratio of barley flour when used instead of cotton seed meal for the cultivation of oyster mushrooms. Mycelial growth was the fastest to 11.4 cm in medium containing 5% barley flour and to 9.0 cm in medium containing 30% barley flour. The mycelial density was slightly lower at 5% and 10% of barley powder addition, but there was no significant difference between the other treatments. The yield of the fruiting bodies was 158 g/850 mL, which was highest on treatment with 10% barley flour as a substitute for cotton seed meal. However, this was lower on treatments with only barley flour than in the control (156 g/850 mL). The diameter of the pileus and the hardness of stipes were highest with 10% barley flour and 10% in cotton seed meal, respectively. The L value was highest with 10% added barley flour, but the a-value and the b-value were not significantly different between treatments. The pH of the mixed media before inoculation with Pleurotus ostreatus was 4.7-5.0, and the total nitrogen content was 1.7-2.2, which was slightly lower than that of the control. The total carbon content was 44.6-45.0, which was not significantly different from that of the control. However, the content of trace elements such as Cu and Fe was lower than those of the control.

Culture of Tricholoma matsutake Mycelium using Solid Matrix (고체 매질을 이용한 송이(松栮)(Tricholoma matsutake)균 배양)

  • Lee, Wi Young;Ahn, Jin Kwon;Ka, Kang Hyeon;Park, Hyun
    • Journal of Korean Society of Forest Science
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    • v.95 no.3
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    • pp.358-364
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    • 2006
  • This study was conducted to develop optimal solid culture medium for Tricholoma matsutake. As the solid matrix, granitic soil, perlite, vermiculate, pine sawdust and peat moss were compared regarding their effected on mycelial growth. Ergosterol content which is a fungal wall component was used as the growth index of the mycelia. Among the various solid matrixes, the granitic soil, perlite and mixture of the two supported the growth most. Barely flour appeared to be very effective on the stimulating of the mycelial growth when added to the solid matrix. An mixture of the matrix contained an even (1:1:1:1, v/v/v/v) mixture of granitic soil, perlite, vermiculate and pine sawdust. T. matsutake started growth 2 weeks after inoculation and reached stationary growth phase after 8th weeks in the solid matrix mixture. The mycelial density in the solid matrix was 7 times higher than that in fairy-ring soil. In addition, 30~70% water content and 10% humus soil in the solid matrix also supported good growth suggesting that T. matsutake needs humus soil for a nutrient sources. The solid matrix developed in the present study could be used to study physiological characteristics of T. matsutake as well.

Correlation between Changes in Microbial/Physicochemical Properties and Persistence of Human Norovirus during Cabbage Kimchi Fermentation

  • Lee, Hee-Min;Lee, Ji-Hyun;Kim, Sung Hyun;Yoon, So-Ra;Lee, Jae Yong;Ha, Ji-Hyoung
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.2019-2027
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    • 2017
  • Recently, cabbage kimchi has occasionally been associated with the foodborne diseases of enteric viruses such as human norovirus (HuNoV). This study aimed to evaluate the correlation between microbial/physicochemical properties and persistence of HuNoV in experimentally contaminated cabbage kimchi fermented and stored at $4^{\circ}C$ or $10^{\circ}C$ for 28 days. Changes in organic acid content, lactic acid bacteria (LAB), acidity, pH, and salinity were analyzed. The recovery of structurally intact HuNoV was examined for up to 28 days post-inoculation, using a NoV GII.4 monoclonal antibody-conjugated immuno-magnetic separation method combined with quantitative real-time reverse transcription polymerase chain reaction. On day 0, LAB loads were $4.70log_{10}$ colony forming units/g and HuNoV GII.4 titers were $2.57log_{10}\;genomic\;copies/{\mu}l$, at both temperatures. After 28 days, intact HuNoV titers decreased to 1.58 ($4^{\circ}C$) and $1.04(10^{\circ}C)log_{10}\;genomic\;copies/{\mu}l$, whereas the LAB density increased. This correlated with a gradual increase in lactic acid and acetic acid at both temperatures. Our findings support a statistical correlation between changes in physicochemical properties and the recovery of structurally intact HuNoV GII.4. Moreover, we determined that the production of organic acid and low pH could affect HuNoV GII.4 titers in cabbage kimchi during fermentation. However, HuNoV GII.4 was not completely eliminated by microbial/physicochemical factors during fermentation, although HuNoV GII.4 was reduced. Based on this, we speculate that the persistence of HuNoV GII.4 may be affected by the continually changing conditions during kimchi fermentation.

Antimicrobial Activity of Hwangryunheadok-tang and Extracts of Scutellariae Radix, Phellodendri Cortex, Coptis Rhizome, and Gardenia Jasminoides against Salmonella Typhimurium (黃連解毒湯, 黃連, 黃芩, 黃柏, 梔子의 Salmonella Typhimurium에 대한 항균효과)

  • Kim, In-soo;Lee, Young-su
    • The Journal of Internal Korean Medicine
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    • v.36 no.2
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    • pp.165-179
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    • 2015
  • Objectives: This experiment investigated the antimicrobial activity of Hwangryunheadok-tang and extracts of Scutellariae radix , Phellodendri cortex , Coptis rhizome , and Gardenia jasminoides against Salmonella typhimurium . Methods: After spreading S. typhimurium on a bacterial culture medium plate, antimicrobial activity was tested by dripping diluted Hwangryunheadok-tang or extracts of Scutellariae radix , Phellodendri cortex , Coptis rhizome , or Gardenia jasminoides (80 μl, diluted 100, 50, 10, and 1%) onto the plate, followed by culture for 16 to 72 hours. The minimal inhibitory concentration (MIC) was tested by dripping the minimum dilution that showed antimicrobial activity (80, 60, 40, and 20 μl) and measuring the density. The antimicrobial activity of Hwangryunheadok-tang and four extracts showed continuous antibacterial activity against S. typhimurium throughout the experiment. Result: 1. S. typhimurium . (Standard Microorganism, ATCC) 1) The Hwangryunheadok-tang and extracts of Scutellariae radix , Phellodendri cortex , and Coptis rhizome showed antibacterial activity in the undiluted solutions and in 50% dilutions. Gardenia jasminoides extract showed potency only in the undiluted solution. The antimicrobial potency of the undiluted solution was increased when the volume of inoculation was increased, but no difference was noted when the culture time was extended. All the extracts showed antimicrobial potency against S. typhimurium . 2. S. typhimurium isolated from diarrhea patients 1) When compared to the standard microorganism experiment on S. typhimurium , the MICs of the five extracts were increased. However, whereas the antimicrobial potency of doxycycline was lost entirely against bacteria isolated from patients with diarrhea, the antimicrobial potency of all the extracts was diminished but did not disappear. 2) The antimicrobial activity of Hwangryunheadok-tang and four extracts was continuous even when the culture time was extended to 16, 24, and 72 hours. Conclusions: The Hwangryunheadok-tang and four kinds of extracts have antimicrobial activity against S. typhimurium . Comparison of a standard microorganism with S. typhimurium isolated from diarrhea patients showed that the antimicrobial activity of all the extracts was better than that of antibiotics. Further studies should focus on the value and benefits of Hwangryunheadok-tang , and the four kinds of extracts as clinical treatments.