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Effect of the Combination of Co-Culture System and Supplemented Protein Sources on the In Vitro Development of Bovine IVF Embryos (각종 공동배양 배지와 첨가 단백질원의 조합이 소 체외수정란의 체외배양에 미치는 영향)

  • Cheong, H.T.;Lee, J.H.;Park, C.K.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.337-345
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    • 1999
  • The present study was conducted to investigate the effects of various co-culture systems and supplemented protein sources on the in vitro development of bovine IVF embryos. Bovine cumulus oocyte complexes (COCs) were matured and fertilized in vitro. Presumptive zygotes with cumulus cells were transferred to TCM-199 or CRlaa containing 10% FBS or 3mg/$m\ell$ BSA, and cultured for 36~40 hr. After primary culture, cleaved embryos were co-cultured with cumulus cells(CC), bovine oviduct epithelial cells(BOEC) or Buffalo rat liver cells (BRLC) in TCM-199 or CRlaa supplemented with FBS or BSA respectively, for further 6 days. Cleavage rate increased with BSA(P<0.01) in the both TCM-199(79%) or CRlaa(74%) When embryos were co-cultured with CC or BOEC in TCM-199, blastocyst development was enhanced with BSA(40% and 43%) compared to FBS (22% and 29%) , whereas in CRlaa no difference observed between BSA(40% and 39%) and FBS (40% and 42%). When embryos were co-cultured with BRLC monolayer, FBS enhanced the blastocyst development (P<0.05) compared to BSA in both TCM-199(41% vs 31%) and CRlaa (44% vs 37%). The result of the present study showed that the cleavage rate of bovine IVF embryos increased with BSA, The result also showed that BSA can enhance the development of IVF embryos in co-culture with CC or BOEC in TCM-199, suggesting the in vitro development is affected by the medium and supplemented protein sources in co-culture with somatic cells.

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Development of a Rapid Enrichment Broth for Vibrio parahaemolyticus Using a Predictive Model of Microbial Growth with Response Surface Analysis (미생물 생장 예측모델과 반응표면분석법을 이용한 Vibrio parahaemolyticus의 신속 증균배지 개발)

  • Yeon-Hee Seo;So-Young Lee;Unji Kim;Se-Wook Oh
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.449-456
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    • 2023
  • In this study, we developed Rapid Enrichment Broth for Vibrio parahaemolyticus (REB-V), a broth capable enriching V. parahaemolyticus from 100 CFU/mL to 106 CFU/mL within 6 hours, which greatly facilitates the rapid detection of V. parahaemolyticus. Using a modified Gompertz model and response surface methodology, we optimized supplement sources to rapidly enrich V. parahaemolyticus. The addition of 0.003 g/10 mL of D-(+)-mannose, 0.002 g/10 mL of L-valine, and 0.002 g/10 mL of magnesium sulfate to 2% (w/v) NaCl BPW was the most effective combination of V. parahaemolyticus enrichment. Optimal V. parahaemolyticus culture conditions using REB-V were at pH 7.84 and 37℃. To confirm REB-V culture efficiency compared to 2% (w/v) NaCl BPW, we assessed the amount of enrichment achieved in 7 hours in each medium and extracted DNA samples from each culture every hour. Real-time PCR was performed using the extracted DNA to verify the applicability of this REB-V culture method to molecular diagnosis. V. parahaemolyticus was enriched to 5.452±0.151 Log CFU/mL in 2% (w/v) NaCl BPW in 7 hours, while in REB-V, it reached 7.831±0.323 Log CFU/mL. This confirmed that REB-V enriched V. parahaemolyticus to more than 106 CFU/mL within 6 hours. The enrichment rate of REB-V was faster than that of 2% (w/v) NaCl BPW, and the amount of enrichment within the same time was greater than that of 2% (w/v) NaCl BPW, indicating that REB-V exhibits excellent enrichment efficiency.

Characterization of Traits Related to Grain Shape in Korean Rice Varieties (국내 육성 벼 품종 입형 관련 특성 분석)

  • Lee, Chang-Min;Lee, Keon-Mi;Baek, Man-Kee;Kim, Woo-Jae;Suh, Jung-Pil;Jeong, Oh-Young;Cho, Young-Chan;Park, Hyun-Su;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.199-213
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    • 2020
  • Grain size and shape are the two important components contributing to rice yield and quality. To analyze traits related to grain-shape, a total of 272 varieties derived from japonica, japonica black and Tongil-type rice accession in Korea were evaluated in this study. The traits, grain length (GL), grain width (GW), grain thickness (GT), length to width ratio (RLW), and 1000-grain weight (TGW) were measured and replicated 10 times. Genes (GW2, GS3, qGL3, qSW5, GS5, TGW6, GW7, and GW8) related to grain-shape were validated in the accessions using specific DNA marker sets. K-mean clustering of the accession based on phenotypic data revealed three groups: group 1 was classified by GW and GT and included most of japonica type, group 2 was classified by RLW and GL reached a medium size and possessed a half spindle-shaped type, and group 3 was classified by TGW, reached a long size and possessed a semi-round shape. In validation tests using the marker sets, both gw2 and tgw6 were validated in less than 1% of the tested accessions and two allelic types, qgl3 and gw8, were only verified in Tongil-type accessions. For GW8 and GW2, any different amplicons were not amplified in any japonica or Tongil-type accessions, respectively. In order to suggest the representative grain-shape gene combinations for each ecotype, the allelic combinations were evaluated by PCR analysis. Cj1 and 2 in japonica (Cj1-7), Cj_b1 and 2 in japonica-black (Cj_b1-3), and CT3 in Tongil-type (CT1-13) turned out to be the dominant combination in each ecotype, respectively. In addition, the results revealed that introgression of four genes (gw2, gs3, qSW5, and GS5) would expand the diversity of grain shape in Korean japonica varieties. The gene combinations information could be utilized practically to understand or enhance grain shape in japonica rice breeding program.

Effects of Activation Regimens of Recipient Cytoplasm, Culture Condition of Donor Embryos and Size of Blastomeres on Development of Reconstituted Bovine Embryos (수핵 난자의 활성화 방법과 공핵 수정란의 배양체계 및 할구의 크기가 소 핵이식 수정란의 발달에 미치는 영향)

  • 심보웅;조성근;이효종;박충생;최상용
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.425-435
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    • 1998
  • To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P<0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.

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Machine learning-based corporate default risk prediction model verification and policy recommendation: Focusing on improvement through stacking ensemble model (머신러닝 기반 기업부도위험 예측모델 검증 및 정책적 제언: 스태킹 앙상블 모델을 통한 개선을 중심으로)

  • Eom, Haneul;Kim, Jaeseong;Choi, Sangok
    • Journal of Intelligence and Information Systems
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    • v.26 no.2
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    • pp.105-129
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    • 2020
  • This study uses corporate data from 2012 to 2018 when K-IFRS was applied in earnest to predict default risks. The data used in the analysis totaled 10,545 rows, consisting of 160 columns including 38 in the statement of financial position, 26 in the statement of comprehensive income, 11 in the statement of cash flows, and 76 in the index of financial ratios. Unlike most previous prior studies used the default event as the basis for learning about default risk, this study calculated default risk using the market capitalization and stock price volatility of each company based on the Merton model. Through this, it was able to solve the problem of data imbalance due to the scarcity of default events, which had been pointed out as the limitation of the existing methodology, and the problem of reflecting the difference in default risk that exists within ordinary companies. Because learning was conducted only by using corporate information available to unlisted companies, default risks of unlisted companies without stock price information can be appropriately derived. Through this, it can provide stable default risk assessment services to unlisted companies that are difficult to determine proper default risk with traditional credit rating models such as small and medium-sized companies and startups. Although there has been an active study of predicting corporate default risks using machine learning recently, model bias issues exist because most studies are making predictions based on a single model. Stable and reliable valuation methodology is required for the calculation of default risk, given that the entity's default risk information is very widely utilized in the market and the sensitivity to the difference in default risk is high. Also, Strict standards are also required for methods of calculation. The credit rating method stipulated by the Financial Services Commission in the Financial Investment Regulations calls for the preparation of evaluation methods, including verification of the adequacy of evaluation methods, in consideration of past statistical data and experiences on credit ratings and changes in future market conditions. This study allowed the reduction of individual models' bias by utilizing stacking ensemble techniques that synthesize various machine learning models. This allows us to capture complex nonlinear relationships between default risk and various corporate information and maximize the advantages of machine learning-based default risk prediction models that take less time to calculate. To calculate forecasts by sub model to be used as input data for the Stacking Ensemble model, training data were divided into seven pieces, and sub-models were trained in a divided set to produce forecasts. To compare the predictive power of the Stacking Ensemble model, Random Forest, MLP, and CNN models were trained with full training data, then the predictive power of each model was verified on the test set. The analysis showed that the Stacking Ensemble model exceeded the predictive power of the Random Forest model, which had the best performance on a single model. Next, to check for statistically significant differences between the Stacking Ensemble model and the forecasts for each individual model, the Pair between the Stacking Ensemble model and each individual model was constructed. Because the results of the Shapiro-wilk normality test also showed that all Pair did not follow normality, Using the nonparametric method wilcoxon rank sum test, we checked whether the two model forecasts that make up the Pair showed statistically significant differences. The analysis showed that the forecasts of the Staging Ensemble model showed statistically significant differences from those of the MLP model and CNN model. In addition, this study can provide a methodology that allows existing credit rating agencies to apply machine learning-based bankruptcy risk prediction methodologies, given that traditional credit rating models can also be reflected as sub-models to calculate the final default probability. Also, the Stacking Ensemble techniques proposed in this study can help design to meet the requirements of the Financial Investment Business Regulations through the combination of various sub-models. We hope that this research will be used as a resource to increase practical use by overcoming and improving the limitations of existing machine learning-based models.

Characteristics and classification of paddy soils on the Gimje-Mangyeong plains (김제만경평야(金堤萬頃平野)의 답토양특성(沓土壤特性)과 그 분류(分類)에 관(關)한 연구(硏究))

  • Shin, Yong Hwa
    • Korean Journal of Soil Science and Fertilizer
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    • v.5 no.2
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    • pp.1-38
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    • 1972
  • This study, designed to establish a classification system of paddy soils and suitability groups on productivity and management of paddy land based on soil characteristics, has been made for the paddy soils on the Gimje-Mangyeong plains. The morphological, physical and chemical properties of the 15 paddy soil series found on these plains are briefly as follows: Ten soil series (Baeggu, Bongnam, Buyong, Gimje, Gongdeog, Honam, Jeonbug, Jisan, Mangyeong and Suam) have a B horizon (cambic B), two soil series (Geugrag and Hwadong) have a Bt horizon (argillic B), and three soil series (Gwanghwal, Hwagye and Sindab) have no B or Bt horizons. Uniquely, both the Bongnam and Gongdeog series contain a muck layer in the lower part of subsoil. Four soil series (Baeggu, Gongdeog, Gwanghwal and Sindab) generally are bluish gray and dark gray, and eight soil series (Bongnam, Buyong, Gimje, Honam, Jeonbug, Jisan, Mangyeong and Suam) are either gray or grayish brown. Three soil series (Geugrag, Hwadong and Hwagye), however, are partially gleyed in the surface and subsurface, but have a yellowish brown to brown subsoil or substrata. Seven soil series (Bongnam, Buyong, Geugrag, Gimje, Gongdeog, Honam and Hwadong) are of fine clayey texture, three soil series (Baeggu, Jeonbug and Jisan) belong to fine loamy and fine silty, three soil series (Gwanghwal, Mangyeong and Suam) to coarse loamy and coarse silty, and two soil series (Hwagye and Sindab) to sandy and sandy skeletal texture classes. The carbon content of the surface soil ranges from 0.29 to 2.18 percent, mostly 1.0 to 2.0 percent. The total nitrogen content of the surface soil ranges from 0.03 to 0.25 percent, showing a tendency to decrease irregularly with depth. The C/N ratio in the surface soil ranges from 4.6 to 15.5, dominantly from 8 to 10. The C/N ratio in the subsoil and substrata, however, has a wide range from 3.0 to 20.25. The soil reaction ranges from 4.5 to 8.0. All soil series except the Gwanghwal and Mangyeong series belong to the acid reaction class. The cation exchange cpacity in the surface soil ranges from 5 to 13 milliequivalents per 100 grams of soil, and in all the subsoil and substrata except those of a sandy texture, from 10 to 20 milliequivalents per 100 grams of soil. The base saturation of the soil series except Baeggu and Gongdeog is more than 60 percent. The active iron content of the surface soil ranges from 0.45 to 1.81 ppm, easily-reduceable manganese from 15 to 148 ppm, and available silica from 36 to 366 ppm. The iron and manganese are generally accumulated in a similar position (10 to 70cm. depth), and silica occurs in the same horizon with that of iron and manganese, or in the deeper horizons in the soil profile. The properties of each soil series extending from the sea shore towards the continental plains change with distance and they are related with distance (x) as follows: y(surface soil, clay content) = $$-0.2491x^2+6.0388x-1.1251$$ y(subsoil or subsurface soil, clay content) = $$-0.31646x^2+7.84818x-2.50008$$ y(surface soil, organic carbon content) = $$-0.0089x^2+0.2192x+0.1366$$ y(subsoil or subsurface soil, pH) = $$-0.0178x^2-0.04534x+8.3531$$ Soil profile development, soil color, depositional and organic layers, soil texture and soil reaction etc. are thought to be the major items that should be considered in a paddy soil classification. It was found that most of the soils belonging to the moderately well, somewhat poorly and poorly drained fine and medium textured soils and moderately deep fine textured soils over coarse materials, produce higher paddy yields in excess of 3,750 kg/ha. and most of the soils belonging to the coarse textured soils, well drained fine textured soils, moderately deep medium textured soils over coarse materials and saline soils, produce yields less than 3,750kg/ha. Soil texture of the profile, available soil depth, salinity and gleying of the surface and subsurface soils etc. seem to be the major factors determining rice yields, and these factors are considered when establishing suitability groups for paddy land. The great group, group, subgroup, family and series are proposed for the classification categories of paddy soils. The soil series is the basic category of the classification. The argillic horizon (Bt horizon) and cambic horizon (B horizon) are proposed as two diagnostic horizons of great group level for the determination of the morphological properties of soils in the classification. The specific soil characteristics considered in the group and subgroup levels are soil color of the profile (bluish gray, gray or yellowish brown), salinity (salic), depositonal (fluvic) and muck layers (mucky), and gleying of surface and subsurface soils (gleyic). The family levels are classified on the basis of soil reaction, soil texture and gravel content of the profile. The definitions are given on each classification category, diagnostic horizons and specific soil characteristics respectively. The soils on these plains are classified in eight subgroups and examined under the existing classification system. Further, the suitability group, can be divided into two major categories, suitability class and subclass. The soils within a suitability class are similar in potential productivity and limitation on use and management. Class 1 through 4 are distinguished from each other by combination of soil characteristics. Subclasses are divided from classes that have the same kind of dominant limitations such as slope(e), wettness(w), sandy(s), gravels(g), salinity(t) and non-gleying of the surface and subsurface soils(n). The above suitability classes and subclasses are examined, and the definitions are given. Seven subclasses are found on these plains for paddy soils. The classification and suitability group of 15 paddy soil series on the Gimje-Mangyeong plains may now be tabulated as follows.

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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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