• Title/Summary/Keyword: free acid

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Free Radical Scavenging Effect and Oxidative Stress Protective Activity of Domestic Processed Polygoni Multiflori Radix (국내산 법제 하수오의 라디칼 소거능 및 산화적 스트레스 개선 효과)

  • Kim, Hyun Young;Kim, Jun Young;Cho, Eun Ju;Choi, Ji Myung;Hwang, Chung Eun;Lee, Hee Yul;Ahn, Min Ju;Lee, Jin Hwan;Kim, Yun-Geun;Ko, Keon Hee;Goo, Young-Min;Oh, Kyeong Yeol;Cho, Kye Man
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.6
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    • pp.809-815
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    • 2015
  • In this study, we confirmed biological compounds from methanol (MeOH) extract of processed Polygoni Multiflori Radix (PPMR), and the radical scavenging effect and oxidative stress protective activity of MeOH extract of PPMR were investigated under in vitro conditions using LLC-$PK_1$ renal epithelial cells. In HPLC analysis, MeOH extract of PPMR contained four species of biological compounds named 2,3,5,4'-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside, emodin, chrysophanol, and rhein. 2,3,5,4'-Tetrahydroxystilbene 2-O-${\beta}$-D-glucoside was detected as the main compound in PPMR as 115.02 mg/kg. MeOH extract of PPMR showed 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) diammonium salt (ABTS), and hydroxyl radical scavenging activities in a concentration- dependent manner. In particular, upon $50{\mu}g/mL$ of PPMR extract treatment, DPPH, ABTS, and hydroxyl radical scavenging activities were approximately 48.4%, 57.9%, and 81.2%, respectively. LLC-$PK_1$ cell viability declined in response to oxidative stress induced by pyrogallol, sodium nitroprusside (SNP), and morpholinosydnonimine (SIN-1) generators of NO, $O_2{^-}$, and $ONOO^-$, respectively. However, MeOH extract of PPMR significantly and dose-dependently inhibited oxidative-stressed LLC-$PK_1$ cell cytotoxicity. In fact, upon $50{\mu}g/mL$ of PPMR extract treatment, LLC-$PK_1$ cell viabilities were approximately 82.1%, 89.1%, and 77.6% compared to stress levels induced by pyrogallol, SNP, and SIN-1, respectively.

Kiwifruit Quality of 'Jecy Gold' as Affected by Soil Types in Jeju Island (제주 토양유형이 '제시골드' 키위 과실의 품질에 미치는 영향)

  • Moon, Doo-Gyung;Kim, Chen-Hwan;Kim, Seong-Cheol;Son, Daniel;Joa, Jae-Ho;Seong, Ki-Cheol;Jung, Hee-Chan;Lim, Han-Cheol;Lee, Young-Jae
    • Journal of agriculture & life science
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    • v.46 no.6
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    • pp.17-24
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    • 2012
  • Soil types for cultivated crops are approximately compose of volcanic ash soils for black(21%) and dark brown soils(41%), and non-volcanic ash soil of red-yellow soil(17%) in Jeju Island. The effects of these soils on fruit qualities of kiwifruit 'Jecy Gold'(Actinidia chinensis cv. Jecy Gold) were investigated in non-heating plastic house. Soil moisture potential was the lowest in the red-yellow soil during fruit growth. However, transverse diameter of fruit in the red-yellow soil was tends to be smaller than in volcanic ash soils, but longitudinal length of fruit was not shown difference by soil types during fruit maturation. Soluble solids in fruit was not differed by soil types until 140 days after of anthesis, after that the red-yellow soil was the highest. No difference on acid contents and hardness of fruit by soil types. Fructose, glucose and sucrose contents in harvested fruit were $4.45{\pm}2.08$, $5.43{\pm}1.13$, and $2.40{\pm}0.40%$ for the red-yellow soil, $2.51{\pm}0.55$, $3.52{\pm}0.86$, and $0.79{\pm}0.33%$ for the black soil and $2.54{\pm}0.47$, $3.52{\pm}0.73$, and $0.73{\pm}0.38%$ for the dark brown soil, respectively. These results show that soluble solid and free sugars in fruit were affected by soil types. It is estimated that soil moisture was rapidly drought in the red-yellow soil of non-volcanic ash soil than in the black and dark brown soils of volcanic ash soil.

Comparison of Physicochemical Properties and Antioxidant Activities of Korean Traditional Kanjang and Garlic Added Kanjang (전통간장과 마늘첨가 간장의 이화학적 특성 및 항산화 활성비교)

  • Shin, Jung-Hye;Kang, Min-Jung;Yang, Seung-Mi;Lee, Soo-Jung;Ryu, Ji-Hyun;Kim, Ra-Jung;Sung, Nak-Ju
    • Journal of agriculture & life science
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    • v.44 no.2
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    • pp.39-48
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    • 2010
  • In order to develop the high quality kanjang in which a functionality is strengthened, the physicochemical properties and antioxidant activities of Korean traditional kanjang and garlic-added kanjang were compared and analyzed. Of color, lightness (L) and redness (a) value were not significant difference between a sample, but yellowness (b) value was higher than garlic-added kanjang. Moisture and crude protein contents were not significantly different, content of crude lipid and ash was higher than traditional kanjang. The pH were 5.02 and 4.91 in traditional and garlic-added kanjang, respectively. The salinity was $20.97{\pm}0.15%$ in garlic-added kanjang, that was significantly lower in garlic-added kanjang. The reducing sugar and total sugar contents were significantly higher in garlic-added kanjang. Na content was occupies 87~89% of total mineral content. The contents of total amino acids were 1,564.02 mg% and 1,932.41 mg% in traditional and garlic-added kanjang, respectiveluy. Also free amino acid was higher in garlic-added kanjang (484.43 mg%) than traditional kanjang (461.13 mg%). The antioxidant activities were increased in a does-dependent manner. In $100{\mu}L/mL$ concentration, electron donating abilities were $14.43{\pm}0.25%$ and $54.6{\pm}1.48%$ in traditional and garlic-added kanjang, ABTs radical scavenging activities of garlic-added kanjang was $57.21{\pm}1.34%$ that was higher than traditional kanjang ($43.27{\pm}0.19%$).

Effects of Phenolic Compounds of Persimmon Leaves on Antioxidative System and Miscellaneous Enzyme Activities Related to Liver Function in Ethanol-Induced Hepatotoxicity of Rats (감잎의 Phenolic Compounds가 에탄올을 투여한 흰쥐의 항산화계 및 기타 효소활성에 미치는 영향)

  • 정창주;윤준식;이명렬
    • Food Science and Preservation
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    • v.11 no.1
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    • pp.79-87
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    • 2004
  • To investigate antioxidative effects of phenolic compounds separated from persimmon leaves(PL)(Diospyros kaki Thunb.) on the ethanol-induced hepatotoxicity in rat, Sprague-Dawley rats weighing 100-150 g were divided into 5 groups; control group(CON), PL(70 mg/kg) administered group(PEl), ethanol(5 mL/kg, 25%) administered group(ETH), PL(70 mg/kg) and ethanol administered group (PE2), and PL(140 mg/kg) and ethanol administered group(PE3), respectively. The antioxidative activity of persimmon leaves decreased in order of ethylacetate>interphase materials>n-butanol>chloroform>n-hexane>water fraction. The growth rate and feed efficiency ratio decreased by ethanol were gradually increased to the adjacent level of CON by administering PL. The serum activities of ALT, alkaline phosphatase and lactic acid dehydrogenase elevated by ethanol were decreased significantly. It was also observed that the activities of SOD, catalase, and GSH-Px of rat liver increased by ethanol were markedly decreased in PL administered group as compared to ETH. The GSH content of liver was decreased by ethanol, but that was increased in PE1 and PE2 compared with ETH as a dose-dependant manner. These results suggested that phenolic compounds separated from persimmon leaves have a possible protective and relievable effect on the ethanol-induced hepatotoxicity in rats.

Effects of Dietary Supplementation of Mulberry Leaves and Silkworm Excreta Extracts on Weight Performance, Blood Characteristics and Cecal Microflorae of Chicks (뽕잎과 잠분 추출물 급여가 육계의 생산성, 혈액 성상 및 맹장 내 미생물군에 미치는 영향)

  • Han, Sang-Hoon;Kim, Dong Wook;Ji, Sang Yoon;Hong, Seong Koo;Kim, Sang-Ho;Lee, Heui-Sam
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.150-160
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    • 2012
  • The object of this study was to evaluate the effects of dietary supplementation of mulberry leaves and silkworm excreta ethanol extracts on weight performances, blood characteristics, cecal microflorae of chickens. Two hundred forty male broiler chicks(Ross) were fed diets for five weeks containing 0.1%(MLA) and 1%(MLB) of mulberry leaves ethanol extracts, and 0.1%(SEA) and 1%(SEB) of silkworm excreta ethanol extracts. Weight performance did show no significant difference in all test groups which were fed with supplementation of mulberry leaves and silkworm excreta ethanol extracts. They showed better weight gain and feed conversion than the negative control group which was fed only with forage without antibiotics. ABTS(2'-azine-bis[3-ethylbenzothiazoline-6-sulfonic acid]) test was conducted to investigate free radical scavenging activity of blood in tested groups. ABTS scavenging activities of tested groups were higher than control groups in significant level, though there was no significant difference(P = 0.396). Specifically, MLB group showed the highest scavenging activity. Blood-level concentration of MDA, which is an indicator of lipid peroxidation, was also decreased in tested groups and the lowest level was observed in SEA(P = 0.001). As storage time increased at $4^{\circ}C$, muscle-level MDA concentrations of all tested groups were generally increased and significant difference was obsereved between tested groups and controls in total increase of MDA concentration($P=4.417{\times}10^{-3}$). In cecal microflorae, SEA and SEB showed decreased total microbe population compared to NC($P=6.462{\times}10^{-5}$) and even to PC. Supplementation of mulberry leave and silkworm excreta ethanol extract did show a similar inhibition effect against Salmonella sp., furthermore, MLB did enhanced the growth of Lactobacillus sp.($P=3.636{\times}10^{-7}$). In summary, ethanol extract of silkworm excreta may be a potential alternative of antibiotics for chicks. In addition, both of ethanol extracts supplementation to broiler chicks would be very useful not only to improve antioxidant effect of blood but also to suppress lipid peroxidation without any loss of weight performance in poultry farming.

Effects of Selenium-Treated Spinacia oleracea L. on Antioxidative Enzyme Activities and Oxidative Damage in Rats Fed High-Fat and High-Cholesterol Diets (셀레늄 강화 시금치가 고지방 및 고콜레스테롤 식이 흰쥐의 항산화 효소활성 및 산화적 손상에 미치는 영향)

  • Song, Won-Yeong;Choi, Jeong-Hwa
    • Journal of Food Hygiene and Safety
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    • v.34 no.4
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    • pp.388-395
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    • 2019
  • The object of the present study was to examine the effect of selenium-treated Spinacia oleracea L. on antioxidative defense system and oxidative damage in rats fed high-fat and high-cholesterol diets. Experimental rats were divided into six groups which were composed of normal diet group (N), high-fat and high-cholesterol diet group (HF), high-fat and high-cholesterol diet with 5% or 10% non-treated spinach supplemented group (SPA or SPB) and high-fat and high-cholesterol diet with 5% or 10% selenium-treated spinach-supplemented group (SSA or SSB). In the antioxidant enzyme activities of hepatic glutathione peroxidase and superoxide dismutase, activities increased in supplementation of non-treated or selenium-treated spinach groups compared to HF group. However, there was no significant difference in the activity of hepatic catalase among all experimental groups. The microsomal superoxide radical content of the SSB group was significantly reduced compared to the HF group. The mitochondrial carbonyl values of the SSB group were significantly reduced compared to the HF group. Thiobarbituric acid reaction substance (TBARS) values in RBC and liver were also reduced in non-treated or selenium-treated spinach-supplemented groups compared to the HF group. The hepatic TBARS values of the supplementation of selenium-treated spinach groups significantly decreased compared to the supplementation of non-treated spinach groups. These results suggest that selenium-treated spinach may reduce oxidative damage by the activation of antioxidative defense system in rats fed high-fat and high-cholesterol diets.

Antioxidant Activities of Peucedanum insolens Kitagawa Root Extracts and Their Anti-inflammatory Effects on LPS-treated RAW264.7 Cells (왕산방풍의 뿌리로부터 제조한 유기용매 분획물에서의 항산화 활성 및 RAW264.7 세포주에서의 항염증 효능)

  • Kim, Jin-Ik;Choi, Yong-Won;Choi, Geun-June;Kang, Ji-An;Lee, In-Young;Narantuya, Nandintsetseg;Oh, Myong-Seok;Cho, Sik-Jae;Moon, Ja-Young
    • Journal of Life Science
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    • v.31 no.1
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    • pp.17-27
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    • 2021
  • This study was performed to investigate the antioxidant activities of subfractions of Peucedanum insolens Kitagawa root in various organic solvents and their anti-inflammatory effects on LPS-treated RAW264.7 cells. First, P. insolens Kitagawa roots were dried at room temperature for one week, chopped, and extracted with 70% ethanol. The resulting extracts were successively sub-fractionated with hexane, chloroform, ethyl acetate, and water. The antioxidant potential of the fractions was evaluated using a DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging assay and by measuring total polyphenol and flavonoid contents. The anti-inflammatory potency of the fractions was evaluated by measuring the inhibition levels of the expressions of inflammatory-mediated genes and proteins (e.g., iNOS, COX-2, IL-1β, and IL-6) in RAW264.7 cells. The results clearly showed that the ethyl acetate fraction of the P. insolens Kitagawa root contained relatively high total flavonoid (34.08±1.68 ㎍ of quercetin equivalents per mg) and total polyphenol (154.1±3.2 ㎍ of gallic acid equivalents per mg) contents. The DPPH assay results showed that the P. insolens Kitagawa root possessed strong free radical scavenging activity in the ethyl acetate fraction. Both the ethyl acetate and hexane fractions showed strong inhibitory potencies to nitric oxide production induced by lipopolysaccharide (1 ㎍/ml) treatment for 24 hr in RAW264.7 cells. The results also showed that both the hexane and ethyl acetate fractions of the P. insolens Kitagawa root strongly inhibited mRNA levels of iNOS, IL-1β, and IL-6, which were overexpressed by LPS treatment for 24 hr in the RAW264.7 cells. These results suggest that P. insolens Kitagawa root may contain compounds that possess strong potency for anti-inflammatory activity. Further studies are needed to discover more detailed modes of action of P. insolens Kitagawa root fractions against inflammation modulation, such as the regulation of cytokine signaling and inflammatory signaling pathways.

A study on γ-Al2O3 Catalyst for N2O Decomposition (N2O 분해를 위한 γ-Al2O3 촉매에 관한 연구)

  • Eun-Han Lee;Tae-Woo Kim;Segi Byun;Doo-Won Seo;Hyo-Jung Hwang;Jueun Baek;Eui-Soon Jeong;Hansung Kim;Shin-Kun Ryi
    • Clean Technology
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    • v.29 no.2
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    • pp.126-134
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    • 2023
  • Direct catalytic decomposition is a promising method for controlling the emission of nitrous oxide (N2O) from the semiconductor and display industries. In this study, a γ-Al2O3 catalyst was developed to reduce N2O emissions by a catalytic decomposition reaction. The γ-Al2O3 catalyst was prepared by an extrusion method using boehmite powder, and a N2O decomposition test was performed using a catalyst reactor that was approximately 25.4 mm (1 in) in diameter packed with approximately 5 mm of catalysts. The N2O decomposition tests were carried out with approximately 1% N2O at 550 to 750 ℃, an ambient pressure, and a GHSV=1800-2000 h-1. To confirm the N2O decomposition properties and the effect of O2 and steam on the N2O decomposition, nitrogen, air, and air and steam were used as atmospheric gases. The catalytic decomposition tests showed that the 1% N2O had almost completely disappeared at 700 ℃ in an N2 atmosphere. However, air and steam decreased the conversion rate drastically. The long term stability test carried out under an N2 atmosphere at 700 ℃ for 350 h showed that the N2O conversion rate remained very stable, confirming no catalytic activity changes. From the results of the N2O decomposition tests and long-term stability test, it is expected that the prepared γ-Al2O3 catalyst can be used to reduce N2O emissions from several industries including the semiconductor, display, and nitric acid manufacturing industry.

Studies on the Physical and Chemical Denatures of Cocoon Bave Sericin throughout Silk Filature Processes (제사과정 전후에서의 견사세리신의 물리화학적 성질변화에 관한 연구)

  • 남중희
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.21-48
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    • 1974
  • The studies were carried out to disclose the physical and chemical properties of sericin fraction obtained from silk cocoon shells and its characteristics of swelling and solubility. The following results were obtained. 1. The physical and chemical properties of sericin fraction. 1) In contrast to the easy water soluble sericin, the hard soluble sericin contains fewer amino acids include of polar side radical while the hard soluble amino acid sach as alanine and leucine were detected. 2) The easy soluble amino acids were found mainly on the outer part of the fibroin, but the hard soluble amino acids were located in the near parts to the fibroin. 3) The swelling and solubility of the sericin could be hardly assayed by the analysis of the amino acid composition, and could be considered to tee closely related to the compound of the sericin crystal and secondary structure. 4) The X-ray patterns of the cocoon filament were ring shape, but they disappeared by the degumming treatment. 5) The sericin of tussah silkworm (A. pernyi), showed stronger circular patterns in the meridian than the regular silkworm (Bombyx mori). 6) There was no pattern difference between Fraction A and B. 7) X-ray diffraction patterns of the Sericin 1, ll and 111 were similar except interference of 8.85A (side chain spacing). 8) The amino acids above 150 in molecular weight such as Cys. Tyr. Phe. His. and Arg. were not found quantitatively by the 60 minutes-hydrolysis (6N-HCI). 9) The X-ray Pattern of 4.6A had a tendency to disappear with hot-water, ether, and alcohol treatment. 10) The partial hydrolysis of sericin showed a cirucular interference (2A) on the meridian. 11) The sericin pellet after hydrolysis was considered to be peptides composed with specific amino acids. 12) The decomposing temperature of Sericin 111 was higher than that of Sericin I and II. 13) Thermogram of the inner portioned sericin of the cocoon shell had double endothermic peaks at 165$^{\circ}C$, and 245$^{\circ}C$, and its decomposing temperature was higher than that of other portioned sericin. 14) The infrared spectroscopic properties among sericin I, II, III and sericin extracted from each layer portion of the cocoon shell were similar. II. The characteristics of seriein swelling and solubility related with silk processing. 1) Fifteen minutes was required to dehydrate the free moisture of cocoon shells with centrifugal force controlled at 13${\times}$10$^4$ dyne/g at 3,000 R.P.M. B) It took 30 minutes for the sericin to show positive reaction with the Folin-Ciocaltue reagent at room temperature. 3) The measurable wave length of the visible radiation was 500-750m${\mu}$, and the highest absorbance was observed at the wave length of 650m${\mu}$. 4) The colorimetric analysis should be conducted at 650mu for low concentration (10$\mu\textrm{g}$/$m\ell$), and at 500m${\mu}$ for the higher concentration to obtain an exact analysis. 5) The absorbing curves of sericin and egg albumin at different wave lengths were similar, but the absorbance of the former was slightly higher than that of the latter. 6) The quantity of the sericin measured by the colorimetric analysis, turned out to be less than by the Kjeldahl method. 7) Both temperature and duration in the cocoon cooking process has much effect on the swelling and solubility of the cocoon shells, but the temperature was more influential than the duration of the treatment. 8) The factorial relation between the temperature and the duration of treatment of the cocoon cooking to check for siricin swelling and solubility showed that the treatment duration should be gradually increased to reach optimum swelling and solubility of sericin with low temperature(70$^{\circ}C$) . High temperature, however, showed more sharp increase. 9) The more increased temperature in the drying of fresh cocoons, the less the sericin swelling and solubility were obtained. 10) In a specific cooking duration, the heavier the cocoon shell is, the less the swelling and solubility were obtained. 11) It was considered that there are differences in swelling or solubility between the filaments of each cocoon layer. 12) Sericin swelling or solubility in the cocoon filament was decreased by the wax extraction.. 13) The ionic surface active agent accelerated the swelling and solubility of the sericin at the range of pH 6-7. 14) In the same conditions as above, the cation agent was absorbed into the sericin. 15) In case of the increase of Ca ang Mg in the reeling water, its pH value drifted toward the acidity. 16) A buffering action was observed between the sericin and the water hardness constituents in the reeling water. 17) The effect of calcium on the swelling and solubility of the sericin was more moderate than that of magnecium. 18) The solute of the water hardness constituents increased the electric conductivity in the reeling water.

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Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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