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Isolation of Marine Bacteria Killing Red Tide Microalgae I. Isolation and Algicidal Properties of Micrococcus sp. LG-1 Possessing Killing Activity for Harmful Dinoflagellate, Cochlodinium polykrikoides (적조생물 살조세균 탐색 I. 유해 적조생물 Cochlodinium Polykrikoides 살조세균 Micrococcus sp. LG-1의 분리와 살조특성)

  • PARK Young-Tae;PARK Ji-Bin;CHUNG Seong-Youn;Song Byung-Chul;LIM Wol-Ae;KIM Chang-Hoon;LEE Won-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.5
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    • pp.767-773
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    • 1998
  • In this study, we have investigated the distributions and killing effects of marine bacteria that tend to kill the red tide microalgae, C. polykikoides in the area of Masan bay from June to October, 1996. To summarize, C. polykikoides killing bacteria were detected at $10^2$ to $10^3$ cells/ml of seawater samples during the survey period, and the bloom was observed in September by containing $4.8\times10^3$cells/ml. It appears however that the number of these bacteria is decreased ($2.0\times10^2$cells/ml) in October, A total of 110 strains were isolated from seawater samples and seawater filtrate (pore size, 0.8 $\mu$m)-containing mixed culture of C. polykikoides in which the mixed culture was grown in f/2 medium. As results we have successfully isolated Micrococcus sp. LG-1 which decreased to less than 10cells/ml within 6days and 5days sfter inoculation of Micrococcus sp. LG-1 into the la9 and logarithmic growth phases of C. polykrikoides respectively. Therefore, it appears that inoculation of Micrococcus sp. LG-1 against the logarithmic C. polykrikoides is more effective than the lag growth phase, (n addition, the killing effects were increased in accordance with bacterial cell densities inoculated in a dose dependent manner. Especially, the filtrate of kitling bacterium culture (nore size, 0.2 $\mu$m) revealed a dramatic effect in which C. polykrikoides were decreased to less than 10 cells/mf of culture within 1 hr, 1,5 hrs, 1,5 hrs, 3.5 hrs. and 5,5 hrs after inoculations of the culture filtrate with concentration of $30\%,\;20\%,\;10\%,\;5\%$ and $2.5\%$, respectively. Moreover Micrococcus sp. LG-1 showed a selective specificity against C. polykrikoides and any other killing effects of Micrococcus sp. LG-1 were not observed against Alexandrium tamarense, Prorocentrum micans, Scrippsiella trochoidea. ana Gymnodinium sanguineum.

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Environmental factors Associated with Disease Development of Garlic White Rot Caused by Two Species of Sclerotium (온도와 토양습도가 마늘 흑색썩음균핵병 발생에 미치는 영향)

  • Kim Yong-Ki;Kwon Mi-Kyung;Shim Hong-Sik;Kim Tack-Soo;Yeh Wan-Hae;Cho Weon-Dae;Choi In-Hu;Lee Seong-Chan;Ko Sug-Ju;Lee Yong-Hwan;Lee Chan-Jung
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.128-134
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    • 2005
  • This study was conducted to elucidate effect of environmental factors on the development of white rot. In order to identify the causal agents causing white rot of Allium crops, we compared DNA profiles of a representative isolate, Sclerotium cepivorum, introduced from foreign country with Korean isolates using UP-PCR. As a result, Sclerotium isolates forming round-shaped sclerotia were identified as Sclerotium cepivorum pertaining in UP-PCR b group and Sclerotium isolates farming anamorphic-shaped sclerotia presumed to be a novel species of Sclerotium based on DNA profiles of UP-PCR. There was a big difference in DNA band pattern between two species of Sclerotium isolated in Korea. Electron micrographs of scanning electron microscope and transmission electron microscope showed morphological differences in sclerotial surface structure and rind layers between two species of Sclerotium. There were more wrinkles and pore spaces on sclerotial surface of Sclerotium sp. forming anamorphic-shaped sclerotia than that of Sclerotium cepivorum forming round-shaped sclerotia. Both of two white rot pathogens grew well at the temperature range of $10-25^{\circ}C$ with optimal temperature of $20^{\circ}C$. Sclerotia of the two pathogens were well formed at $20^{\circ}C$ and well germinated at the temperature range of $20-24^{\circ}C$, Effect of pre-incubation of sclerotia on destruction of sclerotial dormancy of two pathogens was evaluated through storing sclerotia under different temperature condition. The sclerotia of the two pathogens showed an increased capacity to germinate on potato dextroise agar when the sclerotia were incubated for 7 days at $10^{\circ}C$ after pre-treatment at $35^{\circ}C$ for 7 days. At that time, germination rate of Sclerotium sp. and 5. cepivorum was $100\%\;and\;70\%$, respectively. Flooding period and treatment temperature had an effect on sclerotial survival rate of the two pathogens. As flooding period and treatment temperature increased, sclerotial germination rate of the two pathogens decreased. It was confirmed that soil humidity played an important role on development of white rot. It was the highest disease incidence of garlic white rot when garlic were sown at potted soils infested with the two pathogens and adjusted soil humidity to $15\%$ (field moisture capacity, about -300 mb). As soil humidity increase or decrease based on $15\%$ of soil humidity, disease incidence decreased move and more.

Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis (멜론 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.70-82
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    • 2015
  • This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

Isolation of Marine Bacteria Killing Red Tide Microalgae -IV. Characteristics of Algicidal Substances, Produced from Micrococcus sp. LG-5 and the Effects on Marine Organisms- (적조생물 살조세균 탐색 -IV. 살조세균 Micrococcus sp. LG-5가 생산하는 살조물질의 특성과 해양생물에 미치는 영향-)

  • JEONG Seong-Youn;PARK Young-Tae;KIM Mu-Chan;CHOI Seok-Cheol;SEONG Hee-Kyung;KIM Jai-Young;KIM Tae-Un;LEE Won-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.4
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    • pp.339-347
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    • 2000
  • An algicidal bacterium, Micrococcus sp. LG-5 against the harmful dinoflagellate, Cochlodinium polykrikoides was isolated. The optimal conditions for the highest algicidal activity of bacterial culture filtrate showed in the range of $20{\~}30^{\circ}C$, at pH 7.0 and $3.0{\%}$ of NaCl concentration. In addition, $IC_(50)(mean of 50{\%} inhibitory concentration)$ of the culture filtrate against C. polykrikoides after incubation of 5 days was $0.482{\%}$. To investigate heat and pH stability of the culture filtrate of Micrococcus sp. LG-5, the culture filtrate ($pore size, 0.1 {\mu}m$) was heated to $121^{\circ}C for 15 min$ and adjusted pH from 2.0 to 10.0. There were no significant changes in algicidal activity by heat treatment and the pH change between pH from 5.0 to 10.0. The algicidal substances produced from Micrococcus sp. LG-5 were mainly detected in the fraction of $10,000{\~}1,000$ MWCO (molecular weight cut-off). The culture filtrate of Micrococous sp. LG-5 showed antimicrobial activity against Enterococcus faecalis, Escheiichia coli, Uebsiella pneunioniae and Vibrio altinolyticus, but did not show against Pseudomonas aeminosa, P. Buorescens, Salmonella typhi, Staphylococcus aureus, V. cholerae and V parahaemolyicus. The culture filtrate of Micrococcus sp. LG-5 was examined against 16 phytoplankton species and showed the algicidal activity against Ajexandzium tuarense, Eutreptiella Drnnastin, Gymnodinium catenatum, G. mikimotoi, G. sanguineum, eyodinium impuaicum, Heterocapsa triquetra, Heterosipa akashiwo, Prorocentrum micans and Pyraminonas sp.. However no algicidal effects of Micrococcus sp. LG-5 were observed against Chlamydomonas sp., Cylindrotheoa closterium, P. mininum, P. triestimum, Pseudonieschia sp. and Sczipuiella trochoidea. On the other hand, algicidal activity on the tested marinelivefood was not detected except for Isochrysis galbana. In addition, physiological responses of cultured olive flounder (Paralichthys oliraceus) exposed to $1 and 10{\%}$ of the culture filtrate of Micrococcus sp. LG-5 were measured. There were no clear changes in AST, GGT, creatinine, urea, total cholesterol, total protein, albumine, $Mg^(+2), Ca^(+2), Na^+, K^+, and Cl^-$. These results indicate that olive flounders were not affected when they were exposed to the culture filtrate of Micrococcus sp. LG-5.

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