• Research in Plant Disease
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• v.29 no.4
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• pp.345-362
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• 2023

Anthracnose, caused by the Colletotrichum genus, comprises a significant number of plant pathogens and poses a major threat to fruit production worldwide, including South Korea. Colletotrichum species were identified associated with anthracnose in fruits such as apple, persimmon, plum, peach, jujube, walnut, and grape. A polyphasic approach, including morphology, multigene phylogenetics, and pathogenicity testing, was used. Additionally, the in-vitro sensitivity of identified Colletotrichum species to common fungicides was also evaluated. A total of nine Colletotrichum species within two complexes, namely gloeosporioides and acutatum, have been identified as the causal agents of anthracnose in common fruits in South Korea. In the gloeosporioides complex, we found Colletotrichumaenigma, C. fructicola, C. gloeosporioides, C. horii, C. siamense, and C. viniferum. Meanwhile, in the acutatum complex, C. fioriniae, C. nymphaeae, and C. orientalis were identified. Notably, C. fructicola, C. siamense, C. fioriniae, and C. nymphaeae were reported for the first time from apple, C. siamense, C. fioriniae and C. nymphaeae from plum, C. siamense, C. fructicola, and C. fioriniae frompeach, C. siamense and C. horii from persimmon, C. fioriniae from Omija (Schisandra), C. orientalis from walnut, C. nymphaeae from jujube, and C. aenigma, C. fructicola, and C. siamense fromgrape. Fungicide sensitivity tests revealed significant variation in the EC₅₀ values among specific Colletotrichum species when exposed to different fungicides. Moreover, the same Colletotrichum species isolated from different host plants displayed varying sensitivity to the same fungicide.

• Korean Journal of Agricultural Science
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• v.8 no.1
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• pp.51-63
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• 1981

The results of a crossbreeding experiment with a total of 315 litters and 325 pigs of Berkshires, Hampshires, Durocs, Landraces, Large Whites, eight different two-breed crosses and twelve different three-breed crosses, produced at Livestock Experiment Station from 1975 through 1979, are summarized as follows. 1. Number born alive per litter was largest in the D♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ mating, followed by the D♂${\times}$(H♂${\times}$L♀) $F_1$ mating, and smallest in the Hampshires. The pigs in the 3rd-6th parities had larger litter size at birth than those in other parities. 2. Birth weight of pig was heaviest in L♂${\times}$Lw♀ mating and lightest in the Large White. The total litter weight at birth was heaviest in the D♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ mating, followed by D♂${\times}$(H♂${\times}$L♀) $F_1$ ♀ and Lw♂${\times}$L♀ matings, and was smaller in Hampshires and Birkshires. 3. Litter size at weaning was largest in the D♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ mating, followed by D♂${\times}$(H♂${\times}$L♀) $F_1$ ♀ and Lw♂${\times}$L♀ matings, and was smaller in Durocs and Hampshires. The pigs in the 3rd-6th parities had larger litter size at weaning than those in other parities. 4. The total litter weight at weaning was heaviest in the D♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ mating, followed by H♂${\times}$(Lw♂${\times}$L♀) $F_1$♀ and Lw♂${\times}$L♀ matings, and was lighter in Durocs and Hampshires. The weaning weight of pig was largest in D♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ mating and lightest in L♂${\times}$H♀ mating. 5. Survival rate at weaning was highest in L♂${\times}$Lw♀ mating, followed by D♂${\times}$(L♂${\times}$H♀) $F_1$ ♀ and D♂${\times}$(H♂${\times}$L♀) $F_1$ ♀ mating, and was lowest in Durocs. 6. The three-breed cross from D♂${\times}$(H♂${\times}$L♀) $F_1$ ♀ mating had the highest average gain and lowest feed requirement per unit gain, followed by the D♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ and H♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ matings. The Birkshires and Landraces ranked lowest among the 25 mating groups compared for both of the traits. Males had higher average daily gain than females by about 0.06kg and had lower feed requirement by about 0.14. 7. The three-breed crosses from D♂${\times}$(H♂${\times}$L♀) $F_1$ ♀, D♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ and H♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀ matings reached 90kg body weight at younger age than the other groups. The D♂${\times}$(H♂${\times}$L♀) $F_1$ ♀ group reached 90kg at younger age than the Landrace by 39 days. 8. The dressing percentage and lean meat percentage tended to be higher in H♂${\times}$(Lw♂${\times}$L♀) $F_1$ ♀, H♂${\times}$L♀ and L♂${\times}$B♀ matings compared to the other mating groups. The loin-eye area was largest in the Lw♂${\times}$L♀ mating and smallest in the B♂${\times}$L♀ mating. Males had higher dressing percentage, higher lean meat percentage and lion-eye area than females. The backfat was thinnest in purebred Hampshire and was thickest in B♂${\times}$L♀ mating. 9. The results obtained in this study suggest that the two-breed cross from Lw♂${\times}$L♀ mating, and the three-breed crosses from D♂${\times}$(Lw♂${\times}$L-♀) $F_1$ ♀ and D♂${\times}$(H♂${\times}$L♀) $F_1$♀ matings are superior crossbreds for reproductive and pork prodnction performance.

• Korean Journal of Soil Science and Fertilizer
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• v.15 no.1
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• pp.49-50
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• 1982

The objective of this paper is to review the changes in fertilizer use pattern and to discuss some aspects of the fertilizer development in Korea. Fertilizer consumption in Korea have steadily increased to triple the application rates of N, P and K during the 15 years from 1965 to 1980, and Korea became one of the countries which apply fertilizers at the highest rate. The ratio of N: $P_2O_5$: $K_2O$ in fertilizer consumption changed from 55.4 : 31.4 : 13.1 in 1965 to 54.0 : 23.8 : 22.2 in 1980. It can be said that Korean farmers practise a balanced fertilization at least in view of fertilizer consumption as compared to other developing countries. However, differences in soil properties, crops, and climate varying as region were not reflected on fertilization. In the technological development of fertilizer, the chemical form and composition of the fertilizer as well as the suitability to the specific crops must be taken into consideration for the efficient use of fertilizers. Although organic fertilizers and manure are accepted as minor element suppliers, it is necessary to add minor elements into chemical fertilizers on the industrial process. Industrial waste may be used for the agricultural production as a measure of pollution control providing careful study on the waste.

• Korean Journal of Agricultural Science
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• v.2 no.1
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• pp.199-231
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• 1975

It is well known fact that antibiotic residues in milk of cows create significant problem for the fermented dairy industry and public health because of inhibition of starter activity and of creation of allergic disease. It can be assumed that antibiotic residues in milk of other aniimals also can create some problems for their infants as in the case of humen. For the above mentioned reasons, present studies were undertaken to determine concentration and duration of antibiotic residues in milk of cows, goats and dogs following intramuscular or intravenous injection and intramammary infusion of penicillin, streptomycin and oxytetracycline at usual dosage. The cylinder-plate method was used for their assay. The results obtained were summerized as follows: 1) Following the intramuscular injection of penicillin, the antibiotic was detected in milk of cows up to 72 hours, in milk of goats 48 hours and in milk of dogs 60 hours of postinjection. The mean peak concentrations were recorded at 12 hours as 0.136 I.U./ml in cows. 6 hours as 0.773 I.U./ml in goats and 3 hours as 1.192 I.U./ml in dogs. 2) Following the intramuscular injection of streptomycin, the antibiotic was detected in milk of cows and goats up to 36 hours and in milk of dogs 24 hours of post-injection. The mean peak concentration were recorded at 6 hours as $0.26{\mu}g/ml$ in cows and at 3 hours in goats and dogs $0.45{\mu}g/ml$ and $0.36{\mu}g/ml$ respectively. 3) Following the intra venous injection of oxytetracycline, the antibiotic was detectable in milk of all the test animals up to 48 hours of postinjection. The mean peak concentrations were recorded at 6 hours as $3.5{\mu}g/ml$ in cows $2.4{\mu}g/ml$ in goats and $2.0{\mu}g/ml$ in dogs respectively. 4) Following intrarnammary infusion of penicillin in amounts of 100,000 I.U. for cows, 20.000 I.U. for goats and 10,000 I.U. for dogs, the penicillin residues in milk of the infused quarter perssisted to 72 hours in cows and 84 hours in goats and dogs. 5) Following intramammary infusion of streptomycin in amount of 500mg for cows, 100mg for goats and 25mg for dogs, the streptomycin residues in milk of the infused quarter persisted to 72 hours in cows and goats and 60 hours in dogs. 6) Following intramammary infusion of oxytetracycline in amount of 500mg for cows, 100mg for goats and 25mg for dogs, the oxytetracycline residues in milk of the infused quarter persisted to 72 hours in cows and 60 hours in goats and dogs. 7) A corelation between the residual antibiotic concentration and milk yield in cows and goats was observed; That is, the lower in the milk production showed a higher the concentration of an antibiotic residues and a longer the time in persistance. 8) Intramammary transfer of the antibiotic from an infused to non infused quarters, in dogs, was observed following the intramammary infusion of penicillin. streptomycin and oxytetracyclne in amounts of 10.000 I.U. 25mg and 25mg respectively. However, no transfer by 100.000 I.U. or 20.000 I.U. of penicillin. 500mg of streptomycin and 100mg of oxytetracyline was observed in cows and goats. 9) In dogs, minimum dosage of antibiotics for transfer fro in treated to untreated quarters following intramammary infusion were 2,500 I.U. of penicillin and 5mg each of streptomycin and oxytetracycline.

• Korean Journal of Agricultural Science
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• v.19 no.1
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• pp.40-50
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• 1992

This experiment was carried out to stimulate lactic starter culture for yoghurt manufacturing. A each of 1.5% of Bios 2000, CR starter medium, and Yeast extract were added to bulk medium, acidity, pH and changes in the number of lactic acid bacteria were investigated at, intervals of two hours for Lactobacillus bulgaricus and four hours for Streptococus thermophilus and Lactobacillus casei. The results obtained were summarized as follows. 1. The acidity of control arrived at 0.99% after 16 hours of incubation during the incubation of Lactobacillus bulgaricus. Whereas that of CR starter medium reached 1.00% at 12 hours of incubation. Yeast extract, 1.12% at 12 hours, and Bios 2000 reached 0.97% at 10 hours respectively, Thus, Bios 2000 showed the fastest rate of acid production. 2. When the acidity of experiment medium peaked on optimum levels. pH of control was 4.03 in 16 hours of incubation during the incubation of Lactobacillus bulgaricus. Whereas that of Bios 2000 reached 4.10 of Yeast extract reached 3.97 at 12 hours, and of CR starter medium reached 4.05 at 12 hours. 3. Lactic acid bacterial counts were $3.1{\times}10^{10}/ml$ after 16 hours of incubation during the incubation of lactobacillus bulgaricus, Whereas those of Bios 2000 reached $2.1{\times}10^{10}/ml$ at 10 hours, with the fastest stimulation of growth, The counts in CR starter medium were at $2.9{\times}10^{10}/ml$ at 12 hours, and Yeast extract were $3.8{\times}10^{10}/ml$ at 12 hours. 4. The acidity of control, CR starter medium, and Yeast extract reached 0.92% at 44 hours, and 0.96% at 32 hours, and 0.90% at 32 hours respectively, Also, that of Bios 2,000 reached 0.97% at 32 hours, which exhibited the highest, among the treatments. 5. The pH of control was 4.27 at 44 hours. that of CR starter medium was 4.33 at 40 hours and that of Yeast extract was 4.25 at 32 hours during the incubation in Streptococcus thermophilus. Besides, pH of Bios 2000 is lowest as 4.18 at 32 hours. 6. Lactic acid bacterial counts in control, CR starter medium, and Yeast extract during the incubation of Streptococcus thermophilus were $9.8{\times}10^{9}/ml$ at 44 hours,$9.5{\times}10^{8}/ml$ at 40 hours, and $9.6{\times}10^{8}/ml$ at 32 hours. And, the highest number was $2.0{\times}10^{9}/ml$ for Bios 2000 at 32 hours. 7. The acidity of control during the incubation of Lactobacillus casei reached 0.92% at 40 hours, and those of CR starter medium and Yeast extract were 0.95% at 40 hours, and 1.01% at 36 hours respectively. Also, Bios 2000 had the highest acidity as 0.94% at 32 hours. 8. The pH of control, CR starter medium and Yeast extract during the incubation Lactobacillus casei was 4.27 at 40 hours. 4.21 at 40 hours, and 4.15 at 36 hours respectively. Also, Bios 2000 showed the lowest pH, as 4.23, at 32 hours. 9. Lactic acid bacterial counts in control, CR starter medium and Yeast extract during the incubation of Lactobacillus casei were $9.4{\times}10^{7}/ml$ at 40 hours, $1.1{\times}10^{8}/ml$ at 40 hours, and $5.0{\times}10^{8}/ml$ at 36 hours respectively. And, the progress of 32 hours showed the highest number of lactic acid bacteria as$6.4{\times}10^{8}/ml$ in Bios 2000.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.8 no.1
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• pp.129-177
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• 1970

For the purpose of establishing the systematized technical scheme of the cooperative rice cultivation which has most significant impact to improve rice productivity and the farm management, the author have studied the cultivation practices, and the variation of rice growth and yield between the cooperative rice cultivation and the individual rice cultivation at random selected 18 paddy fields. The author also have investigated through comparative method on the cultivation practices, management, organization and operation scheme of the two different rice cultivation methods at 460 paddy fields. The economic feasibility has been ana lysed and added in this report. The results obtained from this study are summarized as follows; 1. In the nursery, the average amount of fertilizer application, especially, phosphate and potassium, and the frequency of chemicals spray for the disease, insect and pest control at the cooperative rice cultivation are significantly higher than those of the individual rice cultivation. 2. The cultivation techniques of the cooperative rice farming after the transplanting can be characterized by a) the earlier transplanting of rice, b) the denser hills per unit area and the lesser number of seedlings per hill, c) the application of larger quantities of fertilizer including nitrogen, phosphate and potassium, d) more divided application of fertilizers, split doses of the nitrogen and potassium, e) the increased frequencies of the chemicals spray for the prevention of disease, insect and pest damages. 3. The rate of lodging in the cooperative rice cultivation was slightly higher than that of the individual rice cultivation, however, the losses of rice yield owing to the occurrence of rice stem borer and grass leaf roller in the cooperative rice cultivation were lower than that of the individual rice cultivation. 4. The culm length, panicle length, straw weight and grain-straw ratio are respectively higher at the cooperative rice cultivation, moreover, the higher variation of the above factors due to different localities of the paddy fields found at the individual rice cultivation. 5. The number of panicles, number of flowers per panicle and the weight of 1, 000 grains, those contributing components to the rice yield were significantly greater in the cooperative rice cultivation, however, not clear difference in the maturing rate was observed. The variation coefficient of the yield component in the cooperative cultivation showed lower than that or the individual rice cultivation. 6. The average yield of brown rice per 10 are in the cooperative rice cultivation obtained 459.0 kilograms while that of the individual rice cultivation brought 374.8 kilograms. The yield of brown rice in the cooperative rice cultivation increased 84.2 kilogram per 10 are over the individual rice cultivation. With lower variation coefficient of the brown rice yield in the cooperative rice cultivation, it can be said that uniformed higher yield could be obtained through the cooperative rice cultivation. 7. Highly significant positive correlations shown between the seeding date and the number of flowers per panicle, the chemical spray and the number of flowers per panicle, the transplanting date and the number of flowers per panicle, phosphate application and yield, potassium application and maturing rate, the split application of fertilizers and yield. Whilst the significant negative correlation was shown between the transplanting date and the maturing rate 8. The results of investigation from 480 paddy fields obtained through comparative method on the following items are identical in general with those obtained at 18 paddy fields: Application of fertilizers, chemical spray for the control of disease, insects and pests both in the nursery and the paddy field, transplanting date, transplanting density, split application of fertilizers and yield n the paddy fields. a) The number of rice varieties used in the cooperative rice cultivation were 13 varieties while the individual rice cultivation used 47 varieties. b) The cooperative rice cultivation has more successfully adopted improved cultivation techniques such as the practice of seed disinfection, adoption of recommended seeding amount, fall ploughing, application of red soil, introduction of power tillers, the rectangular-type transplanting, midsummer drainage and the periodical irrigation. 9. The following results were also obtained from the same investigation and they are: a) In the cooperative rice cultivation, the greater part of the important practices have been carried out through cooperative operation including seed disinfection, ploughing, application of red soil and compost, the control of disease, insects and pests, harvest, threshing and transportation of the products. b) The labor input to the nursery bed and water control in the cooperative rice cultivation was less than that of the individual rice cultivation while the higher rate of labor input was resulted in the red soil and compost application. 10. From the investigation on the organization and operation scheme of the cooperative rice cultivation, the following results were obtained: a) The size of cooperative rice cultivation farm was varied from. 3 ha to 7 ha and 5 ha farm. occupied 55.9 percent of the total farms. And a single cooperative farm was consisted of 10 to 20 plots of paddies. b) The educational back ground of the staff members involved in the cooperative rice cultivation was superior than that of the individual rice cultivation. c) All of the farmers who participated to the questionaires have responded that the cooperative rice cultivation could promise the increased rice yield mainly through the introduction of the improved method of fertilizer application and the effective control of diseases, insects and pests damages. And the majority of farmers were also in the opinion that preparation of the materials and labor input can be timely carried out and the labor requirement for the rice cultivation possibly be saved through the cooperative rice cultivation. d) The farmers who have expressed their wishes to continue and to make further development of the cooperative rice cultivation was 74.5 percent of total farmers participated to the questionaires. 11. From the analysis of economical feasibility on the two different methods of cultivation, the following results were obtained: a) The value of operation cost for the compost, chemical fertilizers, agricultural chemicals and labor input in the cooperative rice cultivation was respectively higher by 335 won, 199 won, 288 won and 303 won over the individual rice cultivation. However, the other production costs showed no distinct differences between the two cultivation methods. b) Although the total value of expenses for the fertilizers, agricultural chemicals, labor input and etc. in the cooperative rice cultivation were approximately doubled to the amount of the individual rice cultivation, the net income, substracted operation costs from the gross income, was obtained 24, 302 won in the cooperative rice cultivation and 20, 168 won was obtained from the individual rice cultivation. Thereby, it can be said that net income from the cooperative rice cultivation increased 4, 134 won over the individual rice cultivation. It was revealed in this study that the cooperative rice cultivation has not only contributed to increment of the farm income through higher yield but also showed as an effective means to introduce highly improved cultivation techniques to the farmers. It may also be concluded, therefore, the cooperative rice cultivation shall continuously renovate the rice production process of the farmers.

• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.67-83
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• 1983

The new microsporidia S80 isolated from, Bombyx mori L. in Korea showed ovoid in the morphology of the spores and the size were measured $2.9{\pm}0.28{\mu}$ in length and $1.7{\pm}0.29{\mu}$ width. No other microsporidian spore like this has not been so far isolated from Silkworm. The length of the polar filament extruded in hydrogen peroxide ($H_2O_2$) at $30^{\circ}C$ was $26{\mu}$ of a round cytoplasm on the top. The spores were partly stained with Giemsa, Safranin-O and Gram as the same staining properties as Nosema bombycis, Microsporidia K 79 and other microsporidian spores. The fine structures were observed under scanning eleceron microscope through ultrathin sectioning. The spore wall was composed of three layers ; the thin exospore of an electron dense rippled layer, the thick electron lucent endospore which was thinning considerably at the polar filament insertion point, and the inner limiting membrane. Polar cap present at the sporeapex, with a long polar filament of 12-13 coils, subtending angle of $60^{\circ}$ to spore axis, which is tubular made up of a multilayered and are a benes core, light ring structure enclosing the dance core, the dark ring structure enclosing the inner light ring structure and the other than and light ring structure bounded from cytoplasm. Lamellate polaroplast occupied the anterior part of the spore, and the two neclei with dense nucleoplasm bounded by a double nuclear envelope were cited in the slight downer middle portion of spore. From the characteristics of the shape, size and fine structures, it is certain to reason the Microsporidia S80 belong to the phylum Microspora, class Microspora, order Microsporida, order Microsporida. The shape of two nuclei cited seems to be genus Nosema, but in the classification for the suborder it should be defined wheather pansporoblasts be formed or not and for the genis especial attempts have been made to define the characters which distinguish the disporous genera in the life cycle. Survey through the infection of the bad cocoons during 1980 to 1982 in South Korea the areas contaminated with new microsporidia were revealed 5 provinces of Kyung-Gi, Kang-Won, Chung-Nam and Chun-Nam. Pathological effects inoculated per os at second instar larvae of silkworm, the LD 50 was $7.1{\times}10^7/ml$ as lower pathogenecity than that of Nosema bombycis Naegeli of $1.2{\times}10_7/ml$. While on the other hand the inoculation of the microsporidia at fourth instar larvae lowerd the whole cocoon weight and cocoon shell weight and significant at 1% level. The microsporidia S80 defined it can not be transmitted transovarially from the result of predictive and collective examination of 21 egg batches from the infected female moth.

• Korean Journal of Weed Science
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• v.9 no.1
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• pp.56-68
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• 1989

aA series of experiments were conducted to investigate the effect of plant growth regulator treatments on the growth and lateral root formation in soybean sprouts in order to establish the effective method of producing root-less or short-rooted soybean sprouts with larger diameter in the hypocotyl. Major results can be summarized as follows. 1. Soybean sprouts showed fairly uniform elongation rate from 3 to g days after imbibition with daily increase of 3.8cm. The speed of elongation of hypocotyl was reduced whereas that of root accelerated 7 days after imbibition. Lateral roots began to emerge fairly evenly from 5 to 9 days after imbibition with a daily increase of 4.4. 2. Auxins(IAA, IBA, NAA, 2,4-D) inhibited hypocotyl elongation and formation of lateral roots and increased hypocotyl diameter without influencing root length and hook diameter at higher concentrations. The dry weight of cotyledon was increased significantly as compared to that of hypocotyl and root. Among the tested auxins, 2, 4-D was the most effective. 3. BA and 4PU-30 significantly reduced elongation of hypocotyl and root and resulted in the biggest diameter of hypocotyl when treated at higher concentrations. The lowest effective concentration of BA to prevent the formation of larval gal roots was 12.5ppm. The formation of lateral roots could be completely prevented by BA and 4PU-30 treatment but kinetin, zeatin, zeatin riboside resulted in many lateral roots and increased thickness of soybean sprouts with little influence. Cotyledon deformation was found in soybean sprouts treated by 4PU-30. 4. 2, 4-D was the most effective for increasing the hypocotyl diameter while 4PU-30 was the most effective for reducing no. of lateral roots. 5. It can be concluded that among the plant growth regulators tested, BA was effective in reducing root length and increasing hypocotyl diameter. BA 12.5 ppm or 15 ppm may thus be the more practical for production of soybean sprouts. 6. ABA showed no significant effect of growth parameter, however ABA 25 ppm inhibited only no of lateral roots with little influence on the growth of seedling. 7. Ethephon inhibited the elongation of hypocotyl and root and increased hypocotyl diameter at higher concentrations. 8. The combined effect of cytokinins and ethephon was very similar to result of BA treatment alone. As the ethephon concentration increased, hypocotyl diameter and dry weight of cotyledon tended to increase.

• Applied Biological Chemistry
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• v.9
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• pp.9-27
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• 1968

Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

• Applied Biological Chemistry
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• v.10
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• pp.69-100
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• 1968

1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.