• Title/Summary/Keyword: inoculation method

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Map based cloning of resistance to bacterial leaf blight gene using QTL analysis in rice

  • Du, Xiao-Xuan;Kim, Kyung-Min
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.138-138
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    • 2017
  • Agriculture is the most primitive civilized Activities of mankind but also the propellant of civilization development. Because it is the most basic material goods source of mankind. Among these materials rice is one of the most important part of these, we call them the substance of survival. From the beginning of the agricultural activities to the present we have experienced three industrial revolutions and are experiencing the Fourth Industrial Revolution. With the development of science and technology makes the efficiency of agricultural production is higher and higher, but compared with the original we are facing the same problem: natural disasters; pests and diseases; now also face the depletion of resources, environmental degradation and other issues. Therefore, improve and cultivate new crop varieties to make it better resistance and more production for better develop modern agriculture. It's very helpful for human social development. And also it is the responsibility and task of modern molecular breeding. In this study, I used bacterial leaf blight to find a better resistance gene to improve the resistance of rice. Frist Cultivate k3 of bacterial leaf blight, than inoculation by leaf clipping method (Kauffman,1973) in CNDH and SNDH population at 40days after rice transplanting. Check the lesion length by inoculation plants at 14days after inoculation, and record data for QTL analysis program. Than I get 4 intervals in 3 different chromosomal regions. I found these defense genes in the 4 intervals. So I used NCBI Justbio, Rapdb, etc. to finding these genes in physical map, than design primer for map base cloning. At last these defense genes will be employed in further research for introduction of the gene to the parental plant and rice breeding for solving food crisis.

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The Effect of Various Concentration of Glucose and Corticosteroid on the Growth of Candida albicans (포도당(葡萄糖) 및 부신피질(副腎皮質)홀몬제(劑)가 Candida albicans의 발육(發育)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Hahm, Jeong-Hee;Kim, Hong-Sik
    • The Korean Journal of Mycology
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    • v.6 no.1
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    • pp.21-27
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    • 1978
  • The growing behavior of Candida albicans in various concentration of glucose and corticosteroid media was studied with the method of modified hanging slide culture. The strains of Candida albicans used in this study were obtained from vaginal swab from outpatients and were isolated from cultured colonies on Sabouraud's glucose agar media. To detect the budding rate of blastospore, the diluted suspension of Candida albicans in normal saline were inoculated into various concentration of glucose (Gl, G2, G3, G4), corticosteroid (S1, S2, S3, S4) and corticosteroid with 10% pepton broth (D1, D2, D3, D4) respectively and cultured at room temperature $(22{\sim}25^{\circ}C)$. The number of budding of blastospore were counted under the high power field of light microscope (400X) at specific time interval, e.g, 1, 2, 3, 6, 9, 12, 18, and 24 hours after inoculation. The results are as following: 1. The most effective budding rate was seen in G4 media (1.25% glucose) in 18 hours aft inoculation (89%). 2. The budding rate in Sabouraud's glucose broth with various concentration of dexamethasone added, was not significantly different from that of simple Sabouraud's glucose broth within 18 hours after inoculation, but there was statistically. significant difference in two budding rate at 24 hours observation. 3. The budding rate in 10% pepton broth media with various concentration of dexamethasone was almost same budding rate in control group, which is normal saline and 10% pepton broth, except on 2 and 24 hours results.

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Circulating Tumor Cell Detection in Lung Cancer Animal Model

  • Chong, Yooyoung;Jung, Yong Chae;Hwang, Euidoo;Cho, Hyun Jin;Kang, Min-Woong;Na, Myung Hoon
    • Journal of Chest Surgery
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    • v.54 no.6
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    • pp.460-465
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    • 2021
  • Background: Metastasis and recurrence of primary cancer are the main causes of cancer mortality. Disseminated tumor cells refer to cancer cells that cause metastasis from primary cancer to other organs. Several recent studies have suggested that circulating tumor cells (CTCs) are associated with the clinical stage, cancer recurrence, cancer metastasis, and prognosis. There are several methods of isolating CTCs from whole blood; in particular, using a membrane filtration system is advantageous due to its cost-effectiveness and availability in clinical settings. In this study, an animal model of lung cancer was established in nude mice using the human large cell lung cancer cell line H460. Methods: Six-week-old nude mice were used. The H460 lung cancer cell line was injected subcutaneously into the nude mice. Blood samples were obtained from the orbital area before cell line injection, 2 weeks after injection, and 2 weeks after tumor excision. Blood samples were filtered using a polycarbonate 12-well Transwell membrane (Corning Inc., Corning, NY, USA). An indirect immunofluorescence assay was performed with the epithelial cell adhesion molecule antibody. The number of stained cells was counted using fluorescence microscopy. Results: The average size of the tumor masses was 35.83 mm. The stained cells were counted before inoculation, 2 weeks after inoculation, and 2 weeks after tumor excision. Cancer cells generally increased after inoculation and decreased after tumor resection. Conclusion: The CTC detection method using the commercial polycarbonate 12-well Transwell (Corning Inc.) membrane is advantageous in terms of cost-effectiveness and convenience.

Observation of Response to PPD Skin Test and Local Side Reactions at Multiple Inoculation Sites after Percutanous Inoculation with BCG Tokyo 172 Strain (경피용 건조 B.C.G. 백신(Tokyo 172주) 접종 후 국소 반응과 투베르쿨린 양전율에 관한 연구)

  • Lee, Jin Soo;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.201-210
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    • 2000
  • Purpose : We observed response to PPD skin test and local side reactions among subjects who received inoculation with Tokyo 172 BCG strain by percutaneous method using multiple puncture device. Methods : 138 infants and young children were enrolled at Yongdong Severance Hospital and 7 private clinics. 5TU PPD skin test were performed at 4 months after inoculation. The local reactions at multiple puncture site were observed in 3 days, 4~6 weeks, 36 weeks, and 48 weeks after inoculations and physical check up was done for evaluation of lymphadenopathy. Results : During 48 weeks of observation period, 96 subjects among 138 who were enrolled were followed up completely with records of PPD skin test and observation of local side reactions, presenting with the photos. The size of the induration after 48 hours of PPD skin test, was less than 5mm in six subjects(6.3%), greater than 10mm in sixty seven subjects(70.0%) and greater than 12mm in forty six subjects(47.9%). All subjects showed inflammatory reaction and pustules at multiple puncture sites and only just small papules, ulcer and pustules remained 4-6 weeks later. Eight to twelve weeks later, all local inflammatory skin reactions disappeared with remaining crust. After 48 weeks, 4(4.2%) subjects showed no scar with only faint stain on the puncture site. More than 70% of subjects showed more than 10 faint pin-point scars on the sites. However, the size of scar was clearly smaller compared to that of intradermal inoculation. There were no cases of lymphadenopathy. Conclusion : We observed good immune response to 5TU PPD skin test among the infant and young children who were immunized with percutanous inoculation of Tokyo 172 BCG strain. We could not find any severe local scar at inoculation sites. A degree of satisfaction of the parents whose children received the percutaneous injection was very high.

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Effect of Temperature, Relative Humidity on Germination and Development of Powdery Mildew (Leveillula taurica) on Pepper and Its Inoculation Method (고추 흰가루병균(Leveillula taurica)의 발아특성 및 접종법)

  • Kim, Dae-Hyun;Park, Jong-Han;Lee, Jung-Sup;Han, Kyung-Sook;Han, You-Kyoung;Hwang, Jeong-Hwan
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.187-192
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    • 2009
  • Pepper powdery mildew, Leveillula taurica is one of the most important pathogens of pepper in the greenhouses and fields in Korea and is becoming a worldwide disease. These experiments were carried out to investigate the optimal germination, disease development conditions, disease incidence and effective inoculation methods of pepper powdery mildew. The incidence of powdery mildew was investigated throughout the country based on the major pepper growing areas in 2009. The average rate of infected leaves ranged from 15.3% to 81.8% in greenhouses and fields. Powdery mildew incidences were more severe in greenhouses than those of fields. The optimal temperature for conidial germination was $25^{\circ}C$ and showed the highest germination at 6 hs after treatment. The range of temperature for germination was $10^{\circ}C$ to $35^{\circ}C$. Temperature of greater than $30^{\circ}C$ and below $20^{\circ}C$ affected the abnormal germination and germ tube elongation. The optimal relative humidity for germination and germ tube elongation was 85% and germination increased as relative humidity increased. Disease development started 8 days after inoculation and showed the highest disease severity at 15 days in greenhouse and field regardless of varieties. Among three different inoculation methods tested the spore dropping method was better than touching with infected leaves and spraying with spore suspension. However, the method has limitation in mass inoculation due to the amount of time consumed.

The Analysis of Management and the Method of Cultivation of Lentinus edodes I. for Full-Development of Mycelium in Bed Logs (표고재배(栽培)의 관리분석(管理分析)과 종균활착(種菌活着)을 위한 골목관리(管理)에 관(關)한 연구(硏究))

  • Joo, Myoung Chil
    • Journal of Korean Society of Forest Science
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    • v.85 no.4
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    • pp.596-604
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    • 1996
  • This study was carried out to offer the successive method of cultivation and increase the productivity of mushroom yield with good quality through the elevation of rate of spawn development for Lentinus edodes. Studied about the analysis of current management of actural cultivation, a base of these, researched and presented for the upward method of productivity through an experiment of the high rate of spawn development and cultivation, putting first cultural environment. The results obtained were as follows ; 1. As the result of the analysis of current management in actural cultivation, many cultivators had a tendency to neglect managements of cultivation. These were reason for the deficiency of labour, funds and the lack of knowledge of cultivation, etc. 2. Water contents in bed logs according to the date of inoculation was shown as the decreasing order of 28.63%(3/12), 25.20%(3/25) and 23.19%(4/10). For the purpose of the maintenance of the water contents, the full-development of mycelium in bed logs and the dispersion of labour, the date of inoculation should be started in the early March. 3. The difference of the rate of spawn development among species was not shown, 100%(Mori 465). 98.98%(Mori 3046) on the spawn in high temperature and 98.97%(Mori 290) on the spawn in low temperature. The relative rate of spawn development was 97.70%(Mori 465), 82.45%(Mori 3046) on the spawn in high temperature and 88.87%(Mori 290) on the spawn in low temperature, it showed the difference. The spawn should be selected carefully in the future, as the spawn of cultivater's preference showed the difference for the development of mycelium. 4. The rate of spawn development following the date of inoculation was 100.0%(3/12), 98.98%(3/25) and 96.79%(4/10) on the spawn in high temperature and 99.09%(3/12), 98.97%(3/25) and 97.89% (4/10) in low temperature, it showed little difference. And the relative rate of spawn development was 97.70%(3/12), 82.45%(3/25) and 81.42%(4/10) on the spawn in high temperature and 93.27%(3/12), 89.67%(3/25) and 88.87%(4/10) that in low temperature, As the result of the relative rate, the time of inoculation of spawn should begin in the early March. 5. The height of stock logs on temporary placing should be less than 60cm at most on the surface, because of the low rate of water contents.

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Characterization of Chitinase in Oak Tissues and Changes in Its Activity Related to Water Stress and Inoculation with Hypoxylon atropunctatum

  • Chun, Se-Chul;Fenn, Patrick;Kim, Kyung-Soo
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.144-151
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    • 1999
  • Chitinase activities from Shumard oak tissues were determined to study changes in chitinase activities related to water stress. The enzyme extracted in sodium acetate buffer (0.1M, pH 4.5) was assayed by a colorimetric method. In addition, the fungal hyphae of Hypoxylon atropunctatum in xylem tissues of oak were observed through scanning electron microscopy. The enzyme in oak tissues was mainly endochitinase, and optimum pH for enzyme activity was 5. Specific chitinase activities from both of stems held under high relative humidity (ranges of 0.63-1.11 pKatal/$\mu\textrm{g}$ of protein) and stems held under low relative humidity (ranges of 0.41-0.99 pKatal/$\mu\textrm{g}$ of protein) were significantly increased following fungal inoculation with H. atropunctatum. However, there was no significant difference in chitinase activities between tissues held under high and low humidities, which might be due to fungal chitinase. Scanning electron microscopy showed holes in fungal hyphae in the xylem tissues of stems held under high humidity but not in the stems held under ow humidity, suggesting that hyphae might be hydrolyzed by plant hydolases such as chitinase.

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Immunohistochemical and Pathological Findings im Mice Inoculated with Encephalomyocarditis Virus (뇌심근염 바이러스 접종 마우스에 대한 병리학적 및 면역조직화학적 소견)

  • 신창호;조성환
    • Korean Journal of Veterinary Service
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    • v.17 no.1
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    • pp.67-80
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    • 1994
  • 8-10 week old ICR mice were infected intracerebrally and intraperitoneally with different encephalomyocarditis virus(K$_3$, $K_{11}$, ATT-VR 129) to observe histopathological and immunohistochemical change. Results obtained throuh the experiments were summarized as follows : 1. No differences in clinical signs by the virus strains and the inoculation routes were found. Mice infected with EMCV showed clinical signs after 3 days of inoculation. Main clinical signs were tremors, convulsions, circling movement, and uni or bilateral hindleg paralysis followed by death on the 3-8 days. In general, most of the infected animals died or recovered closely on the 8th day of postinoculation. 2. At necropsy, petechial and ecchymotic hemorrhages in lung were observed and no specific findings in other were observed. 3. In histopathological observation, neuroal cell degeneration perivascular mononucear cell in-filtration gliosis were appeared in central nervous system. Myocarditis with myocardial degeneration and necrosis, calcification were observed along with acinar cell necrosis of exocrine glands in pancreas, severe glomerulonephritis in kidney. Also, focal necrosis of hepatocytes and interstitial pneymonia hyperemia, hemorrhages in lungs were observed. 4. By immunohistochemical staining using ABCIT method, the positive cells were recognized in intracytoplasm of acinar cell in pancreas and intracytoplasm of neuronal cells in cerebrum.

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Occurrence of Root Rot caused by Fusarium fujikuroi on Adzuki Bean in Korea

  • Min Sun Ha;Hyunjoo Ryu;Sung Kee Hong;Ho Jong Ju;Hyo-Won Choi
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.319-329
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    • 2022
  • In July 2020, wilting symptoms were observed among adzuki bean plants (Vigna angularis var. angularis L.) in the fields in Yeosu, Korea. Infected plants showed yellowing of leaves, browning inside the stems, splitting of stem bark, and wilting. When these plants were uprooted, their roots were found to be brown. The fungal pathogens NC20-737, NC20-738, and NC20-739 were isolated from symptomatic stem and root tissues. These pathogens were identified as a Fusarium fujikuroi species complex based on their morphological characteristics. Molecular identification was performed using the DNA sequence of translation elongation factor 1 alpha and the RNA polymerase II second largest subunit regions. The nucleotide sequences of all three isolates were similar to the F. fujikuroi reference isolates NRRL 13566 and NRRL 5538 of the National Centre for Biotechnology Information GenBank. A pathogenicity test was conducted by the soil inoculation method with cornmeal sand inoculum. Approximately 3 weeks after inoculation, symptoms were observed only in the inoculated adzuki bean seedlings. To the best of our knowledge, this is the first report of Fusarium root rot caused by F. fujikuroi in adzuki beans, both in Korea and worldwide.

Identification and Characteristics of Sclerotinia homoeocarpa Causing Dollar Spot Disease in Zoysiagrass (한국잔디에 발생한 달라스팟 병의 주요 원인균인 Sclerotinia homoeocarpa의 동정 및 특성)

  • Park Dae-Sup;Kim Kyong-Duck;Yeom, Su-Rip;Oh Byung-Seog;Park Byoung-Sun
    • Asian Journal of Turfgrass Science
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    • v.19 no.2
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    • pp.85-94
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    • 2005
  • A fungal isolate was newly collected from Zoysiagrass 'Anyang-Joongji' in small circular patches on a fairway ofa golf course in Korea, which seriously occurred during the early summer period of 2005. The isolate presented on PDAmedium, named Scz1, was closely identical to Sclerotinia homoeocarpa, a casual fungus of dollar spot disease, in cool season turf grasses such as creeping bentgrass. Hereby, this study was accomplished to characterize the isolate and compare it with the fungus, named Scb1, isolated from dollar spot-infected creeping bentgrass (Agrostis palustris Huds. cv Penncross). On PDAmedium, individual mycelial appearance of three isolates was very similar except for the pigment. Mycelial pigments of Scz1 and Scz2 (another analogous isolate collected) were light pinkish on the reverse side of PDA medium but that of Scb1 was dark brownish. In a microscopic study, three isolates were barely distinguishable in the appearance of mycelia. As expected, in the temperaturesensitivity assay, all pathogens were very delicate to $32^{circ}C$ above but not to $30^{circ}C$ below, in which was explained to be one of typical characteristics in S. homoeocarpa. In an artificial inoculation assay, disease symptoms including leaf spots in Zoysiagrass were appeared within 6-7 days after inoculation through the hand inoculation method with the isolate-infested soil. Then the fungus was re-identified from the infected leaf tissues. Interestingly, inoculation of isolate Scz1 gave rise to distinct symptoms in only Zoysiagrass but not in creeping bentgrass 'Penncross' and Kentucky bluegrass 'Midnight'. The observation might be involved in host specific pathogenecity of S. homoeocarpa Scz1 to Zoysiagrass. In a chemical sensitivity assay for the isolate, Scz1, showed a high mycelial inhibition against two fungicides, iprodione and propiconazole. All results described above suggest that S. homoeocarpa Scz1 is a primary pathogen of Zoysia dollar spot disease.