• Journal of Food Hygiene and Safety
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• v.34 no.3
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• pp.227-234
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• 2019

An analytical method was developed for the determination of oxytetracycline in agricultural products using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After the samples were extracted with methanol, the extracts were adjusted to pH 4 by formic acid and sodium chloride was added to remove water. Dispersive solid phase extraction (d-SPE) cleanup was carried out using $MgSO_4$ (anhydrous magnesium sulfate), PSA (primary secondary amine), $C_{18}$ (octadecyl) and GCB (graphitized carbon black). The analytes were quantified and confirmed with LC-MS/MS using ESI (electrospray ionization) in positive ion MRM (multiple reaction monitoring) mode. The matrix-matched calibration curves were constructed using six levels ($0.001{\sim}0.25{\mu}g/mL$) and coefficient of determination ($r^2$) was above 0.99. Recovery results at three concentrations (LOQ, $10{\times}LOQ$, and $50{\times}LOQ$, n=5) were from 80.0 to 108.2% with relative standard deviations (RSDs) less than of 11.4%. For inter-laboratory validation, the average recovery was in the range of 83.5~103.2% and the coefficient of variation (CV) was below 14.1%. All results satisfied the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and the Food Safety Evaluation Department guidelines (2016). The proposed analytical method was accurate, effective and sensitive for oxytetracycline determination in agricultural commodities. This study could be useful for safety management of oxytetracycline residues in agricultural products.

• Journal of Food Hygiene and Safety
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• v.34 no.2
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• pp.124-134
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• 2019

An analytical method was developed for the determination of broflanilide and its metabolites in agricultural products. Sample preparation was conducted using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method and LC-MS/MS (liquid chromatograph-tandem mass spectrometer). The analytes were extracted with acetonitrile and cleaned up using d-SPE (dispersive solid phase extraction) sorbents such as anhydrous magnesium sulfate, primary secondary amine (PSA) and octadecyl ($C_{18}$). The limit of detection (LOD) and quantification (LOQ) were 0.004 and 0.01 mg/kg, respectively. The recovery results for broflanilide, DM-8007 and S(PFP-OH)-8007 ranged between 90.7 to 113.7%, 88.2 to 109.7% and 79.8 to 97.8% at different concentration levels (LOQ, 10LOQ, 50LOQ) with relative standard deviation (RSD) less than 8.8%. The inter-laboratory study recovery results for broflanilide and DM-8007 and S (PFP-OH)-8007 ranged between 86.3 to 109.1%, 87.8 to 109.7% and 78.8 to 102.1%, and RSD values were also below 21%. All values were consistent with the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and the Food and Drug Safety Evaluation guidelines (2016). Therefore, the proposed analytical method was accurate, effective and sensitive for broflanilide determination in agricultural commodities.

• The Korean Journal of Nuclear Medicine Technology
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• v.22 no.2
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• pp.67-73
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• 2018

$[^{11}C]PIB$ synthesis has been performed by a loop-methylation and HPLC purification in our lab. However, this method is time-consuming and requires complicated systems. Thus, we developed an on-cartridge method which simplified the synthetic procedure and reduced time greatly by removing HPLC purification step. We compared 6 different cartridges and evaluated the $[^{11}C]PIB$ production yields and specific activities. $[^{11}C]MeOTf$ was synthesized by using TRACERlab FXC Pro and was transferred into the cartridge by blowing with helium gas for 3 min. To remove byproducts and impurities, cartridges were washed out by 20 mL of 30% EtOH in 0.5 M $NaH_2PO_4$ solution (pH 5.1) and 10 mL of distilled water. And then, $[^{11}C]PIB$ was eluted by 5 mL of 30% EtOH in 0.5 M $NaH_2PO_4$ into the collecting vial containing 10 mL saline. Among the 6 cartridges, only tC18 environmental cartridge could remove impurities and byproducts from $[^{11}C]PIB$ completely and showed higher specific activity than traditional HPLC purification method. This method took only 8 ~ 9 min from methylation to formulation. For the tC18 environmental cartridge and conventional HPLC loop methods, the radiochemical yields were $12.3{\pm}2.2%$ and $13.9{\pm}4.4%$, respectively, and the molar activities were $420.6{\pm}20.4GBq/{\mu}mol$ (n=3) and $78.7{\pm}39.7GBq/{\mu}mol$ (n=41), respectively. We successfully developed a facile on-cartridge methylation method for $[^{11}C]PIB$ synthesis which enabled the procedure more simple and rapid, and showed higher molar radio-activity than HPLC purification method.

• Journal of the Korean Society of Food Science and Nutrition
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• v.16 no.4
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• pp.350-363
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• 1987

This study dealt with the comparison of the individual lipid component and fatty acid composition in the six varieties of dent corn, Zea mays Indentata. The fatty acid and sterol compositions of the total lipid were analyzed by gas liquid chromatography. The total lipid was also fractionated into three lipid classes namely neutral, glyco and phospolipid by the methods of silicic acid column chromatography. The lipid componets of lipid-classes were estimated by thin layer chromatography and TLC-scanner. The contents of total lipid in six varieties of Cdent corn were $3.7{\sim}5.3%$. Total lipid were mainly Composed of triglyceride$(69.8{\sim}75.7%)$ free fatty acid$(13.0{\sim}17.9%)$, lanosterol$(4.8{\sim}6.0%)$. hydrocarbon & esterified sterol$(3.5{\sim}6.0%)$, and polar lipid & pigment$(2.7{\sim}5.9%)$. The contents of triglycerde in $Chech'{\breve{o}}nok$ and Hwangok No.3 were slightly higher then other varieties. The major fatty acid In total lipid from six varieties of dent corn were chiefly consisted of linoleic$(46.0{\sim}61.4%)$, oleic$(21.9{\sim}29.9%)$ and palmitic acid$(10.9{\sim}16.7%)$. Particularly the content of linoleic acid in $Chech'{\breve{o}}nok$ was higher but oleic and palmitic acid in $Chech'{\breve{o}}nok$ were less than other varieties. The compositions of 4-desmethylsterol were mainly composed of siterol $(44.0{\sim}63.2%)$, campestetel$(11.6{\sim}15.5%)$ and stigmasterol$(5.6{\sim}9.1%)$. The content of sitosterol in Chinjuok was higher than other varieties and isofucosterol was detected only in Chinjuok. The compositions of 4-monomethlysterol were mainly composed of obtusifoliol$(17.7{\sim}37.6%)$, gramisterol$(15.0{\sim}27.0%)$ and citrostadienol$(9.1{\sim}17.3%)$. The contents of obtusifoliol and citrostadienol in Kwangok and Chinjuok were less than other varieties. The contents of fractionated neural lipid in Suwon No.19. Kwangok, $Hoengs{\breve{o}}ngok$ and Chinjuok$(90.3{\sim}97.1%)$ were higher than those of $Chech'{\breve{o}}nok$ and Hwangok No.3$(85.5{\sim}86.1%)$. Neutral lipid were mainly composed of triglyceride$(24.7{\sim}80.0%)$, lanosterol$(6.2{\sim}20.2%)$, Cholesterol$(1.0{\sim}50.6%)$, free fatty acid$(4.4{\sim}8.9%)$ and esterified sterol$(1.5{\sim}15.9%)$. The major fatty acid in neutral lipid from six varieties of dent corn were chiefly consisted of linoleic$(26.2{\sim}55.4%)$ oleic$(22.7{\sim}39.1%)$ and palmitic acid$(11.4{\sim}41.6%)$. Particularly the contents of linoleic acid Suwon No.19 and $Chech'{\breve{o}}nok$ were higher but palmtic acid in Suwon No.19 and $Chech'{\breve{o}}nok$ were less tan other varieties. Glycolipd were mainly composed of nlonoglycosflsterol $(17.5{\sim}56.4%)$, monoglycosylce-ramide $(8.2{\sim}25.9%)$ and monoglycoslydiacylglycerol$(12.4{\sim}22.2%)$. The contents of mono-g1ycosrlceramide and monoglycosrlsterol in Chinjuok. Were higher than other varieties. The major fatty acid in glycolipid from six varieties of dent corn were chiefly consisted$(14.6{\sim}39.3%)$, palmitic$(20.0{\sim}26.1%)$, linoleic$(3.6{\sim}26.9%)$ and heptadecanoic acid $(3.3{\sim}24.7%)$. Particutarly the cantents of oleic acid in Chinjuok and heptadecanoic acid in $Chech'{\breve{o}}nok$ were higher than other varieties. Phospholipid were mainly composed of phosphatidyllnositol$(30.9{\sim}86.4%)$ and phosphatidylcholine$(4.5{\sim}22.0%)$. The contents of phosphatidrlinositol in $Hoengs{\breve{o}}ngok$ and Hwanngok No. 3 were less than other varieties. The major fatty acid in phospolipid from six varieties of dent corn chiefly consisted of patmitic$(37.2{\sim}61.6%)$ heptadecanoic$(9.2{\sim}31.8%)$ and oleic acid$(4.3{\sim}17.2%)$. Particuiarlr the content of oleic acid in $Hoengs{\breve{o}}ngok$ was higher but heptadccanoic acid in $Hoengs{\breve{o}}ngok$ was less than othcr varieties.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.6
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• pp.709-720
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• 2009

This study was conducted to identify daily caffeine intakes in beverages for elementary school children and to evaluate its effectiveness after nutrition education. The caffeine contents of 140 commercial beverages were analysed by high performance liquid chromatography-ultraviolet detector (HPLC-UV) and information about their consumption were obtained by surveying 267 children. Researchers gave nutrition education to the children, who were 6 to 11 years old and attended 9 classes of 3 elementary schools, by lecture, Powerpoint file and moving picture. Their preference and intake amount on beverages were investigated by questionnaire before and after nutrition education. The order on caffeine contents was coffee ($33.8{\pm}2.4{\sim}49.1{\pm}5.6\;mg/100\;mL$)> coffee milk ($10.6{\pm}3.3\;mg/100\;mL$)> cola ($6.0{\pm}2.4\;mg/100\;mL$)> green black oolong tea drink ($6.0{\pm}2.4\;mg/100\;mL$)> chocolate milk and chocolate drink ($1.6{\pm}0.7{\sim}1.7\;mg/100\;mL$)> black ice tea mix ($1.3{\pm}1.7\;mg/100\;mL$). The order on children's preference was carbonated drink and fruit and vegetable drink (27%)> sports drink (26%)> processed cocoa mix (7%)> milk (6%)> vitamin & functional drink (3%)> green tea drink (2%)> black tea drink and coffee (1%). The average daily caffeine intakes except tea drink was $5.9{\pm}11.2$ mg/person/day ($0.17{\pm}0.32$ mg/kg bw/day), ranged from $0.0{\sim}80.5$ mg/person/day for children. The sources of caffeine were coffee 57% (3.4 mg/person/day), coffee milk 20% (1.2 mg/person/day), carbonated drink 15% (0.9 mg/person/day), chocolate milk and chocolate drink 6% (0.4 mg/person/day), and vitamin & functional drink 2% (0.1 mg/person/day). After nutrition education, the preference of carbonated drink, coffee, vitamin drinks & functional drink was decreased significantly (p<0.05, p<0.05, p<0.01) and the intakes of carbonated drink, chocolate milk & chocolate drink, and vitamin & functional drink were also decreased significantly (p<0.01, p<0.05, p<0.01). This study has shown that nutrition education influences the preference and the intake behavior of caffeinated beverages.

• Journal of the Korean Wood Science and Technology
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• v.10 no.3
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• pp.13-44
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• 1982

1. Since long time ago, it has been talked about that soaking wood into water for a long time would be profitable for the decreasing of defects such as checking, cupping and bow due to the undue-shrinking and swelling. There are, however, no any actual data providing this fact definitly, although there are some guesses that water soluble extractives might effect on this problem. On the other hand, this is a few work which has been done about the effect of water soluble extractives upon the some physical properties of wood and that it might be related to the above mentioned problem. If man does account for that whether soaking wood into water for a long time would be profitable for the decreasing of defects due to the undue-shrinking and swelling in comparison with unsoaking wood or not, it may bring a great contribution on the reasonable uses of wood. To account for the effect of water soluble extractives upon physical properties of wood, this study has been made at the wood technology laboratory, School of Forestry, Yale university, under competent guidance of Dr. F. F. Wangaard, with the following three different species which had been provided at the same laboratory. 1. Pinus strobus 2. Quercus borealis 3. Hymenaea courbaril 2. The physical properties investigated in this study are as follows. a. Equilibrium moisture content at different relative humidity conditions. b. Shrinkage value from gre condition to different relative humidity conditions and oven dry condition. c. Swelling value from oven dry condition to different relative humidity conditions. d. Specific gravity 3. In order to investigate the effect of water soluble extractives upon physical properties of wood, the experiment has been carried out with two differently treated specimens, that is, one has been treated into water and the other into sugar solution, and with controlled specimens. 4. The quantity of water soluble extractives of each species and the group of chemical compounds in the extracted liquid from each species have shown in Table 36. Between species, there is some difference in quantity of extractives and group of chemical compounds. 5. In the case of equilibrium moisture contents at different relative humidity condition, (a) Except the desorption case at 80% R. H. C. (Relative Humidity Condition), there is a definite line between untreated specimens and treated specimens that is, untreated specimens hold water more than treated specimens at the same R.H.C. (b) The specimens treated into sugar solution have shown almost the same tendency in results compared with the untreated specimens. (c) Between species, there is no any definite relation in equilibrium moisture content each other, however E. M. C. in heartwood of pine is lesser than in sapwood. This might cause from the difference of wood anatomical structure. 6. In the case of shrinkage, (a) The shrinkage value of the treated specimen into water is more than that of the untreated specimens, except anyone case of heartwood of pine at 80% R. H. C. (b) The shrinkage value of treated specimens in the sugar solution is less than that of the others and has almost the same tendency to the untreated specimens. It would mean that the penetration of some sugar into the wood can decrease the shrinkage value of wood. (c) Between species, the shrinkage value of heartwood of pine is less than sapwood of the same, shrinkage value of oak is the largest, Hymenaea is lesser than oak and more than pine. (d) Directional difference of shrinkage value through all species can also see as other all kind of species previously tested. (e) There is a definite relation in between the difference of shrinkage value of treated and untreated specimens and amount of extractives, that is, increasing extractives gives increasing the difference of shrinkage value between treated and untreated specimens. 7. In the case of swelling, (a) The swelling value of treated specimens is greater than that of the untreated specimens through all cases. (b) In comparison with the tangential direction and radial direction, the swelling value of tangential direction is larger than that of radial direction in the same species. (c) Between species, the largest one in swelling values is oak and the smallest pine heartwood, there are also a tendency that species which shrink more swell also more and, on the contrary, species which shrink lesser swell also lesser than the others. 8. In the case of specific gravity, (a) The specific gravity of the treated specimens is larger than that of untreated specimens. This reversed value between treated and untreated specimens has been resulted from the volume of specimen of oven dry condition. (b) Between species, there are differences, that is, the specific gravity of Hymenaea is the largest one and the sapwood of pine is the smallest. 9. Through this investigation, it has been concluded that soaking wood into plain water before use without any special consideration may bring more hastful results than unsoaking for use of wood. However soaking wood into the some specially provided solutions such as salt water or inorganic matter may be dissolved in it, can be profitable for the decreasing shrinkage and swelling, checking, shaking and bow etc. if soaking wood into plain water might bring the decreasing defects, it might come from even shrinking and swelling through all dimension.

• Journal of Korean Society of Forest Science
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• v.32 no.1
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• pp.36-63
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• 1976

The purpose of this study is to elucidate the genetic variation of the natural forest of Pinus densiflora. Three natural populations of the species, which are considered to be superior quality phenotypically, were selected. The locations and conditions of the populations are shown in table 1 and 2. The morphological traits of tree and needle and some other characteristics were presented already in our first report of this series in which population and family differences according to observed characteristics were statistically analyzed. Twenty trees were sampled from each populations, i.e., 60 trees in total. During the autumn of 1974, matured cones were collected from each tree and open-pollinated seeds were extracted in laboratory. Immediately after cone collection, in closed condition, the morphological characteristics were measured. Seed and seed-wing dimensions were also studied. In the spring of 1975, the seeds were sown in the experimental tree nursery located in Suweon. And in the April of 1976, the 1-0 seedlings were transplanted according to the predetermined experimental design, randomized block design with three replications. Because of cone setting condition. the number of family from which progenies were raised by populations were not equal. The numbers of family were 20 in population 1. 18 in population 2 and 15 in population 3. Then, each randomized block contained seedlings of 53 families from 3 populations. The present paper is mainly concerned with the variation of some characteristics of cone, seed, needle, growth performance of seedlings, and chlorophyll and monoterpene compositions of needles. The results obtained are summerized as follows. 1. The meteorological data obtained by averaging the records of 30 year period, observed from the nearest station to each location of populations, are shown in Fig. 3, 4, and 5. The distributional pattern of monthly precipitation are quite similar among locations. However, the precipitation density on population 2, Seosan area, during growing season is lower as compared to the other two populations. Population 1. Cheong-song area, and population 3, Pyong-chang area, are located in inland, but population 2 in the western seacoast. The differences on the average monthly air temperatures and the average monthly lowest temperatures among populations can hardly be found. 2. Available information on the each mother trees (families) studied, such as age, stem height, diameter at breast height, clear-bole-length, crown conditions and others are shown in table 6,7, and 8. 3. The measurements of fresh cone weight, length and the widest diameter of cone are given in Tab]e 9. All these traits arc concerned with the highly significant population differences and family differences within population. And the population difference was also found in the cone-index, that is, length-diameter ratio. 4. Seed-wing length and seed-wing width showed the population differences, and the family differences were also found in both characteristics. Not discussed in this paper, however, seed-wing colours and their shapes indicate the specificity which is inherent to individual trees as shown in photo 3 on page 50. The colour and shape are fully the expression of genetic make up of mother tree. The little variations on these traits are resulted from this reason. The significant differences among populations and among families were found in those characteristics, such as 1000-seed weight, seed length, seed width, and seed thickness as shown in table 11. As to all these dimensions, the values arc always larger in population 1 which is younger in age than that of the other two. The population differences evaluated by cone, seed and seed-wing sizes could partly be attributed to the growth vigorousity. 5. The values of correlation between the characteristics of cone and seed are presented in table 12. As shown, the positive correlations between cone diameter and seed-wing width were calculated in all populations studied. The correlation between seed-wing length and seed length was significantly positive in population 1 and 3 but not in population 2, that is, the r-value is so small as 0.002. in the latter. The correlation between cone length and seed-wing length was highly significant in population 1, but not in population 2. 6. Differences among progenies in growth performances, such as 1-0 and 1-1 seedling height and root collar diameter were highly singificant among populations as well as families within population(Table 13.) 7. The heritability values in narrow sense of population characteristics were estimated on the basis of variance components. The values based on seedling height at each age stage of 1-1 and 1-0 ranged from 0.146 to 0.288 and the values of root collar diameter from 0.060 to 0.130. (Table 14). These heritability values varied according to characteristics and seedling ages. Here what must be stated is that, for calculation of heritability values, the variance values of population was divided by the variance value of environment (error) and family and population. The present authors want to add the heritability values based on family level in the coming report. It might be considered that if the tree age is increased in furture, the heritability value is supposed to be altered or lowered. Examining the heritability values studied previously by many authors, in pine group at age of 7 to 15, the values of height growth ranged from 0.2 to 0.4 in general. The values we obtained are further below than these. 8. The correlation between seedling growth and seed characteristics were examined and the values resulted are shown in table 16. Contrary to our hypothetical premise of positive correlation between 1-0 seedling height and seed weight, non-significance on it was found. However, 1-0 seedling height correlated positively with seed length. And significant correlations between 1-0 and 1-1 seedling height are calculated. 9. The numbers of stomata row calculated separately by abaxial and adaxial side showed highly significant differences among populations, but not in serration density. On serration density, the differences among families within population were highly significant. (Table 17) A fact must be noted is that the correlation between stomata row on abaxial side and adaxial side was highly significant in all populations. Non-significances of correlation coefficient between progenies and parents regarding to stomata row on abaxial side were shown in all populations studied.(Table 18). 10. The contents of chhlorophyll b of the needle were a little more than that of chlorophyll a irrespective of the populations examined. The differences of chlorophyll a, b and a plus b contents were highly significant but not among families within populations as shown in table 20. The contents of chlorophyll a and b are presented by individual trees of each populations in table 21. 11. The occurrence of monoterpene components was examined by gas liquid chromatography (Shimazu, GC-1C type) to evaluate the population difference. There are some papers reporting the chemical geography of pines basing upon monoterpene composition. The number of populations studied here is not enough to state this problem. The kinds of monoterpene observed in needle were ${\alpha}$-pinene, camphene, ${\beta}$-pinene, myrcene, limonene, ${\beta}$-phellandrene and terpinolene plus two unknowns. In analysis of monoterpene composition, the number of sample trees varied with population, I.e., 18 families for population 1, 15 for population 2 and 11 for population3. (Table 22, 23 and 24). The histograms(Fig. 6) of 7 components of monoterpene by population show noticeably higher percentages of ${\alpha}$-pinene irrespective of population and ${\beta}$-phellandrene in the next order. The minor Pinus densiflora monoterpene composition of camphene, myrcene, limonene and terpinolene made up less than 10 percent of the portion in general. The average coefficients of variation of ${\alpha}$-pinene and ${\beta}$-phellandrene were 11 percent. On the contrary to this, the average coefficients of variation of camphene, limonene and terpinolene varied from 20 to 30 percent. And the significant differences between populaiton were observed only in myrcene and ${\beta}$-phellandrene. (Table 25).

• 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.