• Title/Summary/Keyword: in-vitro development

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Antioxidant, Tyrosinase Inhibitory, and Anti-proliferative Activities of Gochujang Added with Cheonggukjang Powder Made from Sword Bean (작두콩 청국장 첨가 고추장의 항산화, tyrosinase 저해 및 암세포 증식 억제 효과)

  • Chang, Moon-Ik;Kim, Jae-Young;Kim, Un-Sung;Baek, Seung-Hwa
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.221-226
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    • 2013
  • This study aimed to examine the antioxidant, tyrosinase inhibitory, and anti-proliferative activities (A549, G361, HT-29, and MDA-MB-231) of fermented gochujang (made from sword bean cheonggukjang powder (SBC) for 90 days. Gochujang was prepared by adding 0 (SBC 0), 2 (SBC 2), 5 (SBC 5), 8 (SBC 8) and 10% (SBC 10) levels with SBC, and all experiments were measured at diluted levels of 20, 50 and 100 times. The antioxidant activity and tyrosinase inhibitory effect demonstrated that SBC 10 increased approximately 1.2 and 1.1 times compared with SBC 0, respectively, at diluted levels of 50 and 100 times. The anti-proliferative effects of A549, G361, and HT-29 presented that SBC 10 were 2.8, 1.1, and 8.9 times higher compared with SBC 0, respectively, at diluted levels of 50, 20, and 100 times. In the case of MDA-MB-231, SBC 10 was 3.7 times higher compared with SBC 5 at diluted level of 20 times. As a result, we confirmed that SBC gochujang was improved for physiological activities and anti-proliferative effects.

Growth Inhibition Effect of Environment-friendly Farm Materials on Fungal Pathogens of Grape (친환경농자재의 포도 진균병 병원균에 대한 생장억제 효과)

  • Kim, Geon-Ju;Choi, Min-Kyung;Park, Jong-Han;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.14 no.3
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    • pp.187-192
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    • 2008
  • Five environment-friendly farm materials including $Chitomate^{(R)}$, $Diegyun^{(R)}$, IC-$66D^{(R)}$, Gold $Bordo^{(R)}$, and $Biospot^{(R)}$ were examined for their growth inhibition effect of the 7 fungal pathogens of grape in vitro. $Diegyun^{(R)}$, being composed of natural ingredients which are extracted from a plant, was the most effective in suppression of mycelial growth of the fungi. $Diegyun^{(R)}$ inhibited the mycelial growth of all of fungi over 75% at $2,500{\mu}g{\cdot}mL^{-1}$ on potato dextrose agar(PDA) except Colletotrichum gloeosporioides 04-159. Growth inhibition effect of $Chitomate^{(R)}$, being composed of the chitosan, varied depending on the fungal pathogens on PDA. It inhibited the mycelial growth of the Botrytis cinerea 06-063 at the rate of 75.8% at $40,000{\mu}g{\cdot}mL^{-1}$ on PDA while it inhibited the mycelial growth of the C. gloeosporioides 04-159 at the rate of 6.5%. IC-$66D^{(R)}$ and Gold $Bordo^{(R)}$ are two different formula of the Bordeaux mixture, showed different control effects on mycelial growth inhibition. Except of Acremonium sp. the growth inhibition of IC-$66D^{(R)}$ was a little higher than Gold $Bordo^{(R)}$. $Biospot^{(R)}$, a chlorine formula, showed the strongest growth inhibition on C. gloeosporioides 04-159 among the farm materials used. Inhibition of spore germination of $Chitomate^{(R)}$, $Biospot^{(R)}$ and Gold $Bordo^{(R)}$ was higher than mycelial growth inhibition for Pseudocercospora vitis 04-152. The results suggest that the different types of environment-friendly farm materials are needed for different disease control in organic grape farm.

Analysis of Semen Parameters, Sperm Activity, and Fertility of Somatic Cell Cloned Hanwoo Bulls (체세포 복제 한우 수소의 정액 성상, 정자의 활동성 및 수정 능력 분석)

  • Bae, Seong-Hoon;Hwang, Seong-Soo;Yang, Byong-Chul;Go, Yeoung-Kyu;Kim, Dong-Hoon;Im, Gi-Sun;Choi, Hwa-Sik;Jin, Dong-Il;Yang, Boh-Suk;Seong, Hwan-Hoo
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.139-143
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    • 2007
  • This study was performed to investigate the reproductive characteristics of the cloned Hanwoo bulls produced by SCNT. The semen ejaculated from the cloned bulls (C-38 and C-39) and normal Hanwoo bull was properly measured the volume, the number of sperm, and the viability of frozen-thawed sperm. The sperm activity was analyzed using computer assisted sperm analysis (CASA). To analyze fertilizing ability of the cloned bulls, in vitro fertilization and artificial insemination were performed using the frozen-thawed semen. There were no differences in semen volume, sperm concentration, and the viability of frozen-thawed sperm between cloned bulls and normal bull. The difference was statistically significant in total motility, curvilinear velocity (VCL), straight-line velocity (VSL), and average-path velocity (VAP) of both cloned bulls compared to those of normal Hanwoo bull, respectively (p<0.05). The cleavage and blastocyst development rate were not different between the groups. five cloned cows were artificially inseminated using the frozen-thawed semen of C-38, two of them became pregnant. Two second generation calves (one male and one female) were produced. Based on these results, the cloned Hanwoo bulls showed normal reproductive abilities of semen parameters and sperm activity to their comparators and produced cloned calves, although there are some individual differences on the parameters.

Antimutagenic Effect and Cytotoxicity to Human Cancer Cell Lines of Colored Potato Extracts (유색감자 추출물의 항돌연변이 및 항암 활성)

  • Park, Young-Eun;Jeong, Jin-Cheol;Cho, Huyn-Mook;Hwang, Young-Sun;Lee, Hyeon-Jin;Choi, Su-San-Na;Lee, Su-Jin;Park, Eun-Sun;Ko, Eun-A;Kim, Nan-Sol;Lim, Jung-Dae;Choung, Myoung-Gun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.1
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    • pp.75-84
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    • 2008
  • Colored potatoes are an excellent source of dietary polyphenols including anthocyanins. Generally, anthocyanins from fruits and vegetables exhibit anti-carcinogenesis and anti-cancer properties in vitro test. This experiment was conducted to know the effects of colored potato extracts contained anthocyanins on antimutagenic activity and anticancer activity to six human cancer cell lines containing LNCaP (androgen-dependent) prostate cancer cells. Extracts of three colored potatoes ('Hongyoung', 'Jayoung' and 'Jasim') and the white potato ('Superior') cultivars were used in this study. The extracts of three colored potatoes inhibited the mutagenicities induced by direct mutagen such as 4-nitro-quinoline-1-oxide (4-NQO) and another indirect mutagens of bezo(a)pyrene (BaP). Also, the extracts of 'Hoyoung' and 'Jayoung' showed higher antimutagenic activity than 'Jasim' and 'Superior' against to direct or indirect mutagen on both strains of TA98 and TA100. The activity of growth-inhibitory of extract of four potato cultivars were screened by SRB (sulphorhodamine B) method on diverse human cancer cells representing different types of cancers. Among the extract of four potato cultivars, the extract of 'Jasim' showed moderate inhibition on proliferation of LNCaP, ACHN and MOLT-4F cells and did not inhibit the proliferation of other cancer cells. On the other hand, extract of 'Superior' did not inhibit the proliferation of any tested cancer cell lines. However, the extracts of 'Hongyoung and Jayoung' inhibited the proliferation of cancer cells with $GI_{50}$ values ranging from 2.5 to $30\;{\mu}g/mL$. On the basis of the $GI_{50}$ values, it is clear that LNCaP cells were more sensitive to extracts of colored potato cultivars than other cancer cells. The extract of 'Jayoung' at $30\;{\mu}g/mL$ were more active and inhibited cell proliferation, and induced apoptosis in LNCaP cells. This result revealed that the extracts of colored potatoes are expected to be good candidate for development into source of antimutagenic and anticancer agent.

Influence of Temperature on the Egg Production and Hatching of Microcotyle sebastis (Monogenea : Microcotylidae), Parasitic on Rockfish, Sebastes schlegeli (수온이 조피볼락에 기생하는 아가미흡충(Microcotyle sebastis)의 산란과 부화에 미치는 영향)

  • Kim, Ki-Hong;Choi, Eun-Seok;Cho, Jae-Bum;Hwang, Yoon-Jung;Park, Soo-Il
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.113-117
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    • 1998
  • The influence of temperature on the rate of egg production and embryonic development of Microcotyle sebastis was investigated to determine the precise time of a second treatment. The survival time of the adults of M. sebastis was inversely proportional to temperature. The number of laid eggs per each replicate during the first 24 h was $39.3{\pm}4.0$ at $10^{\circ}C$, $62.7{\pm}14.2$ at $15^{\circ}C$, $101.0{\pm}5.6$ at $20^{\circ}C$ and $89.0{\pm}11.0$ at $25^{\circ}C$. The time required for egg hatching of M. sebastis was $31.30{\pm}4.88$, $17.52{\pm}3.24$, $11.59{\pm}3.02$ and $10.76{\pm}3.10$ days at 10, 15, 20 and $25^{\circ}C$, respectively. The regression models of the time required for the beginning and 50% point of hatching according to the different temperatures were as follows; Beginning of hatch: D=58.2000-$4.2067{\times}Temp+0.0867{\times}(Temp)^2$ ($P\leq0.01$), 50% of hatch: D=91.3833-$7.5767{\times}Temp+0.1767{\times}(Temp)^2$ ($P\leq0.01$).

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Selection and appropriate culture conditions of antagonistic bacterium Bacillus altitudinis HC7 against button mushroom cobweb disease caused by Cladobotryum mycophilum (양송이버섯 솜털곰팡이병균(Cladobotryum mycophilum)에 대한 길항미생물 Bacillus altitudinis HC7의 선발 및 적정 배양조건)

  • Chan-Jung Lee;Hye-Sung Park;Seong-Yeon Jo;Gi-Hong An;Ja-Yun Kim;Kang-Hyo Lee
    • Journal of Mushroom
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    • v.22 no.2
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    • pp.60-66
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    • 2024
  • This study was conducted to selection and investigate appropriate conditions for mass production of antagonistic microbes to control cobweb disease caused by Cladobotryum mycophilum. A grampositive bacterium was isolated from spent substrate of Agaricus bisporus and showed significant antagonistic activity against Cladobotryum mycophilum. The bacterium was identified as Bacillus altitudinis. based on the cultural, biochemical and physiological characteristics, and 16S rRNA sequence. The isolate is saprophytic, but not parasitic nor pathogenic to cultivated mushroom whereas it showed strong inhibitory effects against C. mycophilum cells in vitro. The control efficacy of B. altitudinis HC7 against cobweb disease of C. mycophilum was up to 78.2% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol system. To define the appropriate conditions for the mass production of the Bacillus altitudinis HC7, we have investigated appropriate culture conditions and effects of various nutrient source on the bacterial growth. The appropriate initial pH and temperature were determined as pH 6.0 and 30℃, respectively. The appropriate concentration of medium elements for the growth of pathogen inhibitor bacterium(Bacillus altitudinis HC7) was determined as follows: 3.0% soluble startch, 10% soytone, 1.0% (NH4)2HPO4, 1.0 mmol KCl, and 0.5% L-asparagine.

Plasma Activity of Lysosomal Enzymes in Active Pulmonary Tuberculosis (활동성 폐결핵 환자에서 혈중 리소솜 효소의 활성도)

  • Koh, Youn-Suck;Choi, Jeong-Eun;Kim, Mi-Kyung;Lim, Chae-Man;Kim, Woo-Sung;Chi, Hyun-Sook;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.5
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    • pp.646-653
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    • 1995
  • Background: The confirmative diagnosis of pulmonary tuberculosis(Tb) can be made by the isolation of Mycobacterium Tuberculosis(MTb) in the culture of the sputum, respiratory secretions or tissues of the patients, but positive result could not always be obtained in pulmonary Tb cases. Although there are many indirect ways of the diagnosis of Tb, clinicians still experience the difficulty in the diagnosis of Tb because each method has its own limitation. Therefore development of a new diagnostic tool is clinically urgent. It was reported that silica cause some lysosomal enzymes to be released from macrophages in vitro and one of these enzymes is elevated in workers exposed to silica dust and in silicotic subjects. In pulmonary Tb, alveolar macrophages are known to be activated after ingestion of MTb. Activated macrophages can kill MTb through oxygen free radical species and digestive enzymes of lysosome. But if macrophages allow the bacilli to grow intracellularly, the macrophages will die finally and local lesion will enlarge. Then it is assumed that the lysosomal enzymes would be released from the dead macrophages. The goal of this investigation was to determine if there are differences in the plasma activities of lysosomal enzymes, ($\beta$-glucuronidase(GLU) and $\beta$-N-acetyl glucosaminidase(NAG), among the groups of active and inactive pulmonary Tb and healthy control, and to see if there is any possibility that the plasma activity of GLU and NAG can be used as diagnostic indicies of active pulmonary Tb. Methods: The plasma were obtained from 20 patients with bacteriologically proven active pulmonary Tb, 15 persons with inactive Tb and 20 normal controls. In 10 patients with active pulmonary Tb, serial samples after 2 months of anti-Tb medications were obtained. Plasma GLU and NAG activities were measured by the fluorometric methods using 4-methylumbelliferyl substrates. All data are expressed as the mean $\pm$ the standard error of the mean. Results: The activites of GLU and NAG in plasma of the patients with active Tb were $21.52{\pm}3.01$ and $325.4{\pm}23.37$(nmol product/h/ml of plasma), respectively. Those of inactive pulmonary Tb were $24.87{\pm}3.78$, $362.36{\pm}33.92$ and those of healthy control were $25.45{\pm}4.05$, $324.44{\pm}28.66$(nmol product/h/ml of plasma), respectively. There were no significant differences in the plasma activities of both enzymes among 3 groups. The plasma activities of GLU at 2 months after anti-Tb medications were increased($42.18{\pm}5.94$ nmol product/h/ml of plasma) in the patients with active pulmonary Tb compared with that at the diagnosis of Tb(P-value <0.05). Conclusion: The results of the present investigation suggest that the measurement of the plasma activities of GLU and NAG in the patients with active pulmonary Tb could not be a useful method for the diagnosis of active Tb. Further investigation is necessary to define the reasons why the plasma activities of the GLU was increased in the patients with active pulmonary Tb after Tb therapy.

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The Mechanism of Interferon-$\gamma$ Induced Cytotoxicity on the Lung Cancer Cell Line, A549 (인터페론감마에 의한 A549 폐암세포주 세포독성의 기전)

  • Oh, Yeon-Mok;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.63-68
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    • 1996
  • Background: Interferon-$\gamma$ has various biologic effects, including antiviral effect, antitumor proliferative effect, activation of macrophage and B lymphocyte, and increased expression of major histocompatibility complex. Especially, antitumor proliferative effect of interferon-$\gamma$ has already been proved to be important in vivo as well as in vitro. And, clinical studies of interferon-$\gamma$ have been tried in lung cancer patients. However, the mechanism of antitumor effect of interferon-$\gamma$ has not yet been established despite of many hypotheses. "Necrosis" is a type of cell death which is well known to occur in the circumstances of severe stresses. In contrast, "apoptosis" is another type of cell death which occurs in such biological circumstances as embryonic development, regression of organs, and self-tolerance of lymphocytes. And, apoptosis is an active process of cell death in which cells are dying with fragmentations of their cytoplasms and nuclei. And, in the process of apoptosis the DNAs of cells are cleaved between nucleosomes by unidentified endonuclease and therefore DNAs of apoptotic cells result in a typical electrophoresis pattern known as DNA ladder pattern. Recently it has been suggested that cytotoxic effect of interferon-$\gamma$ occurs via apoptosis. To elucidate the mechanism of antitumor cytotoxic effect of interferon-$\gamma$, we microscopically observed a lung cancer cell line, A549 which was treated with interferon-$\gamma$. We observed A545 treated with interferon-$\gamma$ was dying fragmented. And so, we performed this study to find out that the mechanism of antitumor cytotoxic effect of interferon-$\gamma$ be apoptosis. Method: We treated A549, human lung cancer cell line with various concentration of interferon-$\gamma$ and quantified its cytotoxic effect of various periods, 24 hours, 72 hours and, 120 hours by MTT(dimethylthiazolyl diphenyltetrazolium bromide) bioassay. Also, after we treated A549 with 100 units/mi of interferon-$\gamma$ for 120 hours, we observed the pattern of cell death with inverted microscope and we extracted DNAs from the dead A549 cells and observed the pattern of 1.5% agarose gel electrophoresis with ethidium bromide staining. Result: 1) Cytotoxic effect of interferon-$\gamma$ on A549: For the first 24 hours, threre was little cytotoxic effect and for between 24 hours and 72 hours, there was the beginning of cytotoxic effect and for 120 hours there was increased cytotoxic effect. 2) Pattern of A549 cell death by interferon-$\gamma$: We observed with inverted microscope that A549 cells were dying fragmented. 3) DNA ladder pattern of gel electrophoresis: We observed DNA ladder pattern of gel electrophoresis of extracted DNAs from dead A549 cells. Conclusion: We concluded that the mechanism of interferon-$\gamma$induced cytotoxicity on lung cancer cell line, A549 be via apoptosis.

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Fermentation Characteristics of Juice Pomace Feed by Horse Feces Microbes (말분변 미생물을 이용한 주스박 사료의 발효 특성)

  • Hwang, Won-Uk;Kim, Gyeom-Heon;Niu, Kai-Min;Lim, Joung-Ho;Woo, Jae-Hoon;Chae, Hyun-Seok;Park, Nam-Geon;Kim, Soo-Ki
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.4
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    • pp.290-300
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    • 2017
  • In vitro fermentation was conducted to figure out alternative fiber sources for horse feed. For the development of value-added products as a horse feed resource, the pomaces from apple, carrot, grape, and citrus were fermented under solid-state conditions in the presence of 60% soybean meal with 40% of each fruit pomace at 60% of moisture content. Lactobacillus plantarum SK3873, Lactobacillus plantarum SK3893, Weissella cibaria SK3880, and Bacillus subtilis SK3889 were isolated from the fermented fruit pomace by inoculation of horse feces. For the growth of Bacillus subtilis, Saccharomyces cerevisiae, and Lactobacillus plantarum, they were inoculated in 3-step order at 0, 12, and 24 h, respectively. The fruit pomace was fermented for 48 h at $35^{\circ}C$. The pH of the apple, carrot, grape, citrus and all mixed pomaces decreased from 5.45~6.25 to 4.40~4.77. Microbial growth was maintained at $10^8{\sim}10^9cfu/g$. After 12 and 24 h incubation, dry matter of carrot pomace were highest at 54.84 and 56.66%, respectively (P<0.05) and that of grape pomace was lower than others during fermentation (P<0.05). Dry matter was generally reduced by about 20%. NDF decreased gradually or maintained after 24 h, indicating the fiber degradation. Ash content tended to decrease during fermentation. After 48 hours fermentation, Bacillus, yeast and Lactobacillus showed an excellent growth by using juice by-products. These results suggest that fermented juice pomace has a potential as horse feedstuff with probiotics to maintain beneficial microflora in horse gut.

Ecological Studies on the Transition of Sheath Blight of Rice in Korea (한국(韓國)에서의 벼 잎집무늬마름병 발생변동(發生變動)에 관(關)한 생태학적(生態學的) 연구(硏究))

  • Yu, Seung-hun
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.283-316
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    • 1977
  • In an attempt to obtain a basic information to develop an effective integrated system of controlling sheath blight of rice in Korea, the transition of this disease, the variation of cultural characters and pathogenicity of the pathogen, environmental conditions affecting the disease outbreak and varietal resistance have been investigated. 1. Rice sheath blight which has been minor disease in the past was widely spread, especially since 1971. This disease has promptly spread all over the country and infected 65.2% of total rice growing area in 1976. Various factors are considered to be related to such transition of this disease. Above all, increace of application of nitrogenous fertilizer, early season and earlier cultivation of rice, introduction of more susceptible "Tongil" varieties etc. must be important factors influencing the outbreak of this disease. 2. Great variations in cultural characteristics-such as mycelial growth rate, color of the medium, amount of the aerial mycelium, shape and color of the sclerotia- and in the pathogenicity of isolates of the pathogen, Thanatephorus cucumeris Dank were observed. The optimum temperature for mycelial growth also varied with isolates, from $25^{\circ}C$ to $30^{\circ}C$. There were not necessarily any correlation between curtural characteristics and pathogenicity of isolates of Thanatephorus cucumens. 3. Mycelial grow th of isolates of Thanatephorus cucumens on the PDA medium were correlated with the air temperatures of the region where the isolates were collected. The isolates from the regions with high temperature grew well on PDA medium at $35^{\circ}C$ than those from the region with low temperature, on the other hand, the isolates from the regions with the low temperature grew well on the same medium at $12^{\circ}C$ than those from the regions with high temperature. 4. Pectin polygalacturonase (PG) and cellulase (Cx) were most active on the 3rd day after inoculation on the leaves of rice plant with Thanatephorus cucumeris, whereas pectin methylestrase (PE) was most active on the 4th day after inoculation. Relationship between the activities of PE of isolates and the strength of pathogenicity of isolates was obtained, but PG and cellulase activities were not correlated with pathogenicity of isolates. 5. The tolerence of sclerotia from in-vitro culture to low temperature varied with their water content, the dried cultural sclerotia were more tolerent than wet ones, Dried cultural sclerotia maintained almost 100% germinability for 45 days at $-20^{\circ}C$, whereas wet sclerotia lost viability at $-5^{\circ}C$. The germination ratio of the sclerotia after overwintering changed from 18% to 70% according to the water content of the test paddy fields and the ratio was low in wet paddy condition. 6. To investigate the host range of this fungi in and near paddy field, 17 weeds were inoculated with fungi. The lesions of sheath blight disease was obserbed on Sagittaria trifolia L., Echinochloa crusgalli P. Beauv., Monochoria vaginal is Presl, Polygonum Hydropiper L., Eclipta prostrata L., Digitaria sanguinalis Scapoli. 7. When the level of nitrogen applied was doubled over standard level, total nitrogen content in rice sheath increased, ami when silicate was applied, starch content in rice sheath decreased, inducing the rice plants more susceptible to sheath blight disease. Increased dressing of potash ferilizer reduced the incidence of sheat blight disease. 8. The percentage of infected stems in the early period increased more in the narrow hill plot than in the wide hill plot, but in the late period this tendency was inversed; the percentage of infected stems as well as severity in the wide hill plot increased more compared to the narrow hill plot, and the disease severity in the one plant per hill plot was also low. The number of stems in the wide hill plot was more than the number of stems in the narrow hill plot. This indicates that the microclimate, such as the relative humidity, in the narrow hill plot was more favorable for the development of this disease. 9. There was a high negative correlation between the disease severity of varieties to the sheath blight and the maturity of the varieties, that is, the early varieties were more susceptible than the late ones, and much-tillering varieties usually showed more infection than less tillering varieties. 10. No relationship was obtained between the percentage of infected stems in the early period and the severity after heading, whereas a distinct relationship was obtained between former and latter after Aug. 10.

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