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In Vitro Fertilization of Pig Oocytes Matured In­Vitro by liquid Boar Spermatozoa (체외성숙 돼지 난포란의 액상정액을 이용한 체외수정)

  • 박창식;이영주
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.17-23
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    • 2002
  • The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.

The Variation of Natural Population of Pinus densiflora S. et Z. in Korea (VIII) - Genetic Variation of the progeny originated from Injye, Jeongsun, and Samchuk Populations - (소나무 천연집단(天然集團)의 변이(變異)에 관(關)한 연구(硏究) - 인제(麟蹄), 정선(旌善), 삼척집단(三陟集團)의 차대(次代)의 유전변이(遺傳變異) -)

  • Yim, Kyong Bin;Lee, Kyong Jae
    • Journal of Korean Society of Forest Science
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    • v.43 no.1
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    • pp.20-30
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    • 1979
  • The purpose of present study is to analyze the genetic variation of natural stand of Pinus densiflora. In 1976 following after the seletion of 1974 and 1975, twenty trees from each of three natural populations of the species were selected and seeds were collected, and the locations and conditions of populations are presented in table 1, 2 and figure 1. Some morphological traits of the populations were already detailed in our fifth report of this series. The morphological traits of cone, seed and seed-wing, and also the growth performances and needle characters of the seedling were observed in the present study according to the previous methods. The results obtained are summarized as follows; 1. The meteorological data obtained by averaging the records of 30 year period(1931~1960) measured from the nearest meteorological station to each population are shown in fig. 2, 3, 4. The distributional patterns of investigated climate factors are generally considered to be similar among the locations. However, the precipitation density during growing season and the air temperature during dormant season on Samchuk area (Pop. 9) were quite different from those of the other areas. 2. The measurements of fresh cone weight, length, diameter and cone index (i.e.: length to diameter ratio) are presented in table 7. As shown in table 7, all these traits except for cone diameter seem to be not significant in population and to be highly significant in family differences within population. 3. The morohological traits of seed and seed-wing are detailed in table 8, 9, and highly significant differences are recognized among the populations and the families within population in seed weight, seed length, seed thickness but not among the populations in the other observed traits. The values of correlation between the characteristics of cone and seed are presented in table 12. As shown, the correlation between cone length and seed wing length, between seed wing width and seed width were significantly positive in population 8 and 9 but in population 7. The positive correlations between seed length and seed width were calculated in all populations studied 4. Significant statistical differences among populations and families within population are observed in the growth performances of 1-0 seedling height of these progenies. But the differences in 1-1 seedling height and root collar diameter are shown only among familes within population. As shown in table 13, the most parts of correlations are not significant statistically between the growth performances of seedling and the seed characters. 5. As shown in table 15, statistical differences are considered to be significant among the populations in stomata row on both sides of the needle but not in serration density. The correlations between progenies and parents are not generally observed in the investigated traits of needle as shown in table 16.

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The Variation of Natural Population of Pinus densiflora S. et Z. in Korea (VI) - Genetic Variation of the Progency Originated from Myong-Ju, Ul-Jin and Suweon Populations - (소나무 천연집단(天然集團)의 변이(變異)에 관(關)한 연구(硏究)(VI) - 명주(溟洲), 울진(蔚珍), 수원(水原) 소나무 집단(集團)의 차대(次代)의 유전변이(遺傳變異) -)

  • Yim, Kyong Bin;Kwon, Ki Won;Lee, Kyong Jae
    • Journal of Korean Society of Forest Science
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    • v.38 no.1
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    • pp.33-45
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    • 1978
  • The purpose of present study is to analyze the genetic variation of natural stand of Pinus densiflora. In 1975 following after the selection of 1974, twenty trees from each of three natural populations of the species were selected and their open-pollinated seeds were collected, and the locations and conditions of the populations ate presented in table 1, 2 and figure 1. Some morphological traits of the populations were already detailed in our second report of this series, in which Myong-Ju and Ul-Jin populations were regarded to be superior phenotypically to suweon population. The morphological traits of cone, seed and seed-wing, and also the growth performances and needle characters of the seedling were observed in the present study according to the previous methods. The results obtained are summarized as follows; 1. The meteorological data obtained by averaging the records of 30 year period (1931~1960) measured from the nearest meteorological stations to each population are shown in fig.2, 3, 4. The distributional patterns of investigated climate factors are generally considered to be similar among the locations. However, the precipitation density during growing season and the air temperature during dormant season on Suweon area, population 6, were quite different from those of the other areas. 2. The measurements of fresh cone weight, length, diameter and cone index, i.e., length to diameter ratio are presented in table 7. As shown in table 7, all these traits except for cone diameter seem to be highly significant in population differences and family differences within population. 3. The morphological traits of seed and seed-wing are detailed in table 8, 9, and highly significant differences are recognized among the populations and the families within population in seed-wing length, seed-wing index, seed weight, seed-length and seed index but not among the populations in the other observed traits. The values of correlation coefficient between the characters of cone and seed are given in table 10 and the positive significant correlations can be observed in the most parts of the compared traits. 4. Significant statistical differences among populations and families within population are observed in the growth performances of 1-0 and 1-1 seedling height of these progenies. But the differences in root collar diameter are shown only among families within population. As shown in table 13, the most parts of correlations are not significant statistically between the growth performances of seedling and the seed characters. 5. The number of stomata row on both sides of needle and the serration density were measured in the seedlings from each of the families of the three populations. As shown in table 15, statistical differences are considered to be significant among the populations and among the families within population in serration density but not among the populations in stomata row on both sides of the needle. The results differ from those of the third report of this series. Even if one of the reason seems to be the diversity of selected populations, it could not be confirmed definitely. The correlations between progenies and parents are not generally observed in the investigated traits of needle as shown in table 16.

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Studies on a Factor Affecting Composts Maturity During Composting of SWine Manure (돈분 퇴비화 중 부숙도에 미치는 영향인자 구명)

  • Kim, T.I.;Song, J. I.;Yang, C.B.;Kim, M.K.
    • Journal of Animal Science and Technology
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    • v.46 no.2
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    • pp.261-272
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    • 2004
  • This study was conducted to investigate indices affecting composts maturity for swine manure compost produced in a commercial composting facility with air-forced from the bottom. The composting was made of swine manure mixed with puffing rice hull(6: 4) and turned by escalating agitator twice a day. Composting samples were collected periodically during a 45-d composting cycle at that system, showing that indices of Ammonium-N to Nitrate-N ratio were sensitive indicators of composting quality. Pile temperature maintained more than 62$^{\circ}C$ and water contents decreased about 20% for 25days of composting. A great variety and high numbers of aerobic thermophilic heterotropic microbes playing critical roles in stability of composts have been examined in the final composts, sbowing that they were detected $10^8$ to $10^{10}$ $CFUg^{-1}$ in mesophilic bacteria, $10^3$ - $10^4$ in fungi and $10^6$ - $10^8$ in actinomycetes, respectively. The results of this study for detennining a factor affecting compost stability evaluations based on composting steps were as follows; 1. Ammonium-N concentrations were highest at the beginning of composting, reaching approximately 421mg/kg. However Ammonium-N concentrations were lower during curing, reaching approximately l04mg/kg just after 45 day. The ratio between $NH_4-N$ and $NO_3-N$ was above II at the beginning of composting and less than 2 at the final step(45 day). 2. Seed germination Index was dependent upon the compost phytotoxicity and its nutrition. The phytotocity caused the GI to low during the period of active composting(till 25 days of composting time) depending on the value of the undiluted. After 25 days of composting time, the GI was dependent upon compost nutrition. The Gennination index of the final step was calculated at over 80 without regard to treatments. 3. E4: E6 ratio in humic acid of composts was correlatively decreased from 8.86 to 6.76 during the period of active composting. After 25 days of composting time, the E4: E6 was consistently decreased from 6.76 to 4.67($r^2$ of total composting period was 0.95). 4. Water soluble carbon had a tendency to increase from 0.54% to 0.78%during the period of active composting. After 25 days of composting time, it was consistently decreased from 0.78% to 0.42%. Water soluble nitrogen increased from 0.22% to 0.32% during the period of 15 days after initial composting while decreased from 0.32% to 0.21% after 15days of composting. In consequence, the correlation coefficient($r^2$) between water soluble carbon and water soluble nitrogen was 0.12 during the period of active composting mule was 0.50 after 25 days of composting time

Studies on the fate of nitrogen in the paddy soil (답토양(沓土壤)에서 질소(窒素)의 동태(動態)에 관(關)한 연구(硏究))

  • Kim, Kwang Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.9 no.1
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    • pp.17-23
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    • 1976
  • In order to investigate the fate of nitrogen in the paddy soil, Suchang, Hwasoon and Susan soil which have different properties, were treated with several nitrogen fertilizers such as ammonium chloride, ammonium sulfate, urea and SCU (sulfur-coated urea), and incubated under water-logged condition in $30^{\circ}C$ incubator. $NH_4-N$, $NO_3-N$, $Fe^{++}$ and pH in soil and stagnant water, were determined at 10, 20, 30, 40 and 50 days after incubation. The obtained results were summarized as follows: 1. The effect of rising temperature was increased in order of Hwasoon>Suchang>Susan and the effect of air drying soil was risen in order of Susan>Hwasoon>Suchang, while the rate of ammonication was in order of Susan>Suchang>Hwasoon. 2. The changes of $NH_4-N$ in stagnant water was dependent upon the nitrogen concentration of $NH_4Cl$ and $(NH_4)SO_4$ plat was high and decreased after 30 days incubation, but increased after 40 days and then decreased again. In contrast with the above, $NH_4-N$ concentration of urea and SCU plot was low but the change showed slightly through the incubation period. 3. Accumulation of $NH_4-N$ in the oxidative layer of the $NH_4Cl$ and $(NH_4)_2SO_4$ plot was higher than that of urea and SCU plot and $NH_4-N$ content was decreased with the incubation period. The change of $NH_4-N$ in the reductive layer showed the same pattern. 4. The changes of $NO_3-N$ in the stagnant water were different according to soil properties and nitrogen fertilizer. $NO_3-N$ concentration in stagnant water of urea and SCU plot was higher than in the $NH_4-Cl$ $(NH_4)_2SO_4$ plot and nearly disappeared after 30 to 40 days incubation. 5. The $NO_3-N$ concentration in the oxidative layer of soil was higher than reductive layer. The pattern of change was different in accordance with soil properties and nitrogen fertilizers. In general, nitrification in urea and SCU plot was more increased than $(NH_4)_2SO_4$ plot. In reductive layer, the concentration of $NO_3-N$ was very low until 30 days incubation and thereafter increased slightly. 6. Upon the concentration of $NH_4-N$ and $NO_3-N$ in stagnant water and soil, it was assumed that denitification of urea and SCU plot was higher than $NH_4Cl$ and $(NH_4)_2SO_4$ plot and denitrified nitrogen in incubation period was above 50%.

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Studies on the Consumptine Use of Irrigated Water in Paddy Fields During the Growing of Rice Plants(III) (벼생유기간중의 논에서의 분석소비에 관한 연구(II))

  • 민병섭
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.11 no.4
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    • pp.1775-1782
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    • 1969
  • The results of the study on the consumptine use of irrigated water in paddy fields during the growing season of rice plants are summarized as follows. 1. Transpiration and evaporation from water surface. 1) Amount of transpiration of rice plant increases gradually after transplantation and suddenly increases in the head swelling period and reaches the peak between the end of the head swelling poriod and early period of heading and flowering. (the sixth period for early maturing variety, the seventh period for medium or late maturing varieties), then it decreases gradually after that, for early, medium and late maturing varieties. 2) In the transpiration of rice plants there is hardly any difference among varieties up to the fifth period, but the early maturing variety is the most vigorous in the sixth period, and the late maturing variety is more vigorous than others continuously after the seventh period. 3) The amount of transpiration of the sixth period for early maturing variety of the seventh period for medium and late maturing variety in which transpiration is the most vigorous, is 15% or 16% of the total amount of transpiration through all periods. 4) Transpiration of rice plants must be determined by using transpiration intensity as the standard coefficient of computation of amount of transpiration, because it originates in the physiological action.(Table 7) 5) Transpiration ratio of rice plants is approximately 450 to 480 6) Equations which are able to compute amount of transpiration of each variety up th the heading-flowering peried, in which the amount of transpiration of rice plants is the maximum in this study are as follows: Early maturing variety ; Y=0.658+1.088X Medium maturing variety ; Y=0.780+1.050X Late maturing variety ; Y=0.646+1.091X Y=amount of transpiration ; X=number of period. 7) As we know from figure 1 and 2, correlation between the amount evaporation from water surface in paddy fields and amount of transpiration shows high negative. 8) It is possible to calculate the amount of evaporation from the water surface in the paddy field for varieties used in this study on the base of ratio of it to amount of evaporation by atmometer(Table 11) and Table 10. Also the amount of evaporation from the water surface in the paddy field is to be computed by the following equations until the period in which it is the minimum quantity the sixth period for early maturing variety and the seventh period for medium or late maturing varieties. Early maturing variety ; Y=4.67-0.58X Medium maturing variety ; Y=4.70-0.59X Late maturing variety ; Y=4.71-0.59X Y=amount of evaporation from water surface in the paddy field X=number of period. 9) Changes in the amount of evapo-transpiration of each growing period have the same tendency as transpiration, and the maximum quantity of early maturing variety is in the sixth period and medium or late maturing varieties are in the seventh period. 10) The amount of evapo-transpiration can be calculated on the base of the evapo-transpiration intensity (Table 14) and Tablet 12, for varieties used in this study. Also, it is possible to compute it according to the following equations with in the period of maximum quantity. Early maturing variety ; Y=5.36+0.503X Medium maturing variety ; Y=5.41+0.456X Late maturing variety ; Y=5.80+0.494X Y=amount of evapo-transpiration. X=number of period. 11) Ratios of the total amount of evapo-transpiration to the total amount of evaporation by atmometer through all growing periods, are 1.23 for early maturing variety, 1.25 for medium maturing variety, 1.27 for late maturing variety, respectively. 12) Only air temperature shows high correlation in relation between amount of evapo-transpiration and climatic conditions from the viewpoint of Korean climatic conditions through all growing periods of rice plants. 2. Amount of percolation 1) The amount of percolation for computation of planning water requirment ought to depend on water holding dates. 3. Available rainfall 1) The available rainfall and its coefficient of each period during the growing season of paddy fields are shown in Table 8. 2) The ratio (available coefficient) of available rainfall to the amount of rainfall during the growing season of paddy fields seems to be from 65% to 75% as the standard in Korea. 3) Available rainfall during the growing season of paddy fields in the common year is estimated to be about 550 millimeters. 4. Effects to be influenced upon percolation by transpiration of rice plants. 1) The stronger absorbtive action is, the more the amount of percolation decreases, because absorbtive action of rice plant roots influence upon percolation(Table 21, Table 22) 2) In case of planting of rice plants, there are several entirely different changes in the amount of percolation in the forenoon, at night and in the afternoon during the growing season, that is, is the morning and at night, the amount of percolation increases gradually after transplantation to the peak in the end of July or the early part of August (wast or soil temperature is the highest), and it decreases gradually after that, neverthless, in the afternoon, it decreases gradually after transplantation to be at the minimum in the middle of August, and it increases gradually after that. 3) In spite of the increasing amount of transpiration, the amount of daytime percolation decreases gadually after transplantation and appears to suddenly decrease about head swelling dates or heading-flowering period, but it begins to increase suddenly at the end of August again. 4) Changs of amount of percolation during all growing periods show some variable phenomena, that is, amount of percolation decreases after the end of July, and it increases in end August again, also it decreases after that once more. This phenomena may be influenced complexly from water or soil temperature(night time and forenoon) as absorbtive action of rice plant roots. 5) Correlation between the amount of daytime percolation and the amount of transpiration shows high negative, amount of night percolation is influenced by water or soil temperature, but there is little no influence by transpiration. It is estimated that the amount of a daily percolation is more influenced by of other causes than transpiration. 6) Correlation between the amount of night percoe, lation and water or soil temp tureshows high positive, but there is not any correlation between the amount of forenoon percolation or afternoon percolation and water of soil temperature. 7) There is high positive correlation which is r=+0.8382 between the amount of daily percolation of planting pot of rice plant and amount and amount of daily percolation of non-planting pot. 8) The total amount of percolation through all growin. periods of rice plants may be influenced more from specific permeability of soil, water of soil temperature, and otheres than transpiration of rice plants.

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Studies on Neck Blast Infection of Rice Plant (벼 이삭목도열병(病)의 감염(感染)에 관(關)한 연구(硏究))

  • Kim, Hong Gi;Park, Jong Seong
    • Korean Journal of Agricultural Science
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    • v.12 no.2
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    • pp.206-241
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    • 1985
  • Attempts to search infection period, infection speed in the tissue of neck blast of rice plant, location of inoculum source and effects of several conditions about the leaf sheath of rice plants for neck blast incidence have been made. 1. The most infectious period for neck blast incidence was the booting stage just before heading date, and most of necks have been infected during the booting stage and on heading date. But $Indica{\times}Japonica$ hybrid varieties had shown always high possibility for infection after booting stage. 2. Incubation period for neck blast of rice plants under natural conditions had rather a long period ranging from 10 to 22 days. Under artificial inoculation condition incubation period in the young panicle was shorter than in the old panicle. Panicles that emerged from the sheath of flag leaf had long incubation period, with a low infection rate and they also shown slow infection speed in the tissue. 3. Considering the incubation period of neck blast of rice plant, we assumed that the most effective application periods of chemicals are 5-10 days for immediate effective chemicals and 10-15 days for slow effective chemicals before heading. 4. Infiltration of conidia into the leaf sheath of rice plant carried out by saturation effect with water through the suture of the upper three leaves. The number of conidia observed in the leaf sheath during the booting stage were higher than those in the leaf sheath during other stages. Ligule had protected to infiltrate of conidia into the leaf sheath. 5. When conidia were infiltrated into the leaf sheath, the highest number of attached conidia was observed on the panicle base and panicle axis with hairs and degenerated panicle, which seemed to promote the infection of neck blast. 6. The lowest spore concentration for neck blast incidence was variable with rice varietal groups. $Indica{\times}Japonica$ hybrid varieties were infected easily compared to the Japonica type varieties, especially. The number of spores for neck blast incidence in $Indica{\times}Japonica$ hybrid varieties was less than 100 and disease index was higher also in $Indica{\times}Japonica$ hybrid than in Japonica type varieties. 7. Nitrogen content and silicate content were related with blast incidence in necks of rice plants in the different growing stage changed during growing period. Nitrogen content increased from booting stage to heading date and then decreased gradually as time passes. Silicate content increased from booting stage after heading with time. Change of these content promoted to increase neck blast infection. 8. Conidia moved to rice plant by ascending and desending dispersal and then attached on the rice plant. Conidia transfered horizontally was found very negligible. So we presumed that infection rate of neck blast was very low after emergence of panicle base from the leaf sheath. Also ascending air current by temperature difference between upper and lower side of rice plant seemed to increase the liberation of spores. 9. Conidial number of the blast fungus collected just before and after heading date was closely related with neck blast incidence. Lesions on three leaves from the top were closely related with neck blast incidence, because they had high potential for conidia formation of rice blast fungus and they were direct inoculum sources for neck blast. 10. The condition inside the leaf sheath was very favorable for the incidence of neck blast and the neck blast incidence in the leaf sheath increased as the level of fertilizer applied increased. Therefore, the infection rate of neck blast on the all panicle parts such as panicle base, panicle branches, spikelets, nodes, and internodes inside the leaf sheath didn't show differences due to varietal resistance or fertilizers applied. 11. Except for others among dominant species of fungi in the leaf sheath, only Gerlachia oryzae appeared to promote incidence of neck blast. It was assumed that days for heading of varieties were related with neck blast incidence.

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Agronomical studies on the major environmental factors of rice culture in Korea (수도재배의 주요환경요인에 관한 해석적 조사연구)

  • Yung-Sup Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.3
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    • pp.49-82
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    • 1965
  • For the stable and high yields of low-land rice in Korea, the characteristics of rice plant for the vegetative and physiological responses, plant type formation, and yield components have been studied in order to obtain the fundamental data for the improvement of cultural practices, especially for the ideal fertilizer application. Furthermore the environmental conditions in Korea including temperatures, light, precipitation, and soil conditions have been compared in the broad sense with those in Japan, and the application of nitrogen, phosphorus, potassium, silicate and other micro-nutrients were described in relation to the characteristics of environmental conditions for the improvement of fertilizer application. 1. The average yield of polished-rice per 10 are in Korea is about 204 kg and this values are much less than those in Japan and Taiwan where they produce 77% to 13% more than in Korea. The rate of yield increase a year in Korea is 4.2 kg, but in Japan and Taiwan the rates of yield increase a year are 81 % and 62%, respectively. It was also found that the coefficient of variation of yield is 7.7% in Korea, 6.7% in Japan and 2.5% in Taiwan. This means that the stability of producing rice in Korea is very low when compared with those in Japan and Taiwan. 2. It was learned from the results obtained from the 'annual yield estimation experiment' that there are big differences in the respect of plant type formations between rice crops grown in Japan and Korea. The important differences found were as follows: (1) The numbers of spikelets per 3.3 square meters are 891 in Korea and 1, 007 in Japan(13% more than in Korea). (2) The numbers of tillers per 3.3 square meters at the stage of maximum tillering are 1, 150 in Korea, but in Japan they showed 19% more than in Korea. (3) The ratio of effective tillers to total tillers is 77.5% in Korea and 74.7% in Japan, which seems to be higher in Korea than in Japan. But the ratio in Korea is very low when considered the numbers of total tillers in both countries. (4) The ratio of grain to straw is 85.4% in Korea and 96.3% in Japan. 3. The average temperatures during the growing season at the area of Suwon, Kwangjoo and Taegu are almost same as those in the district of Jookokoo(Fookoo yama) in Japan, i.e., the temperatures during the rice-growing season in Korea are similar to those in the southern-warm regions of Japan. 4. Considering the minimum temperatures at the stage of limiting transplanting, 13$^{\circ}C$, the time of transplanting might be 30 to 40 days earlier than presently practicing transplanting time, which comes around June 10. 5. The temperatures during the vegetative growth in Korea were higher than those temperatures that needed in the protein synthesis which ate the main metabolism during this stage. However, the temperatures at the time of reproductive growth was lower than the temperatures that needed in the sugar assimilation which is main metabolism in this stage. In this point of view, it might be considered that the proper time of growing rice plant in Korea would be rather earlier. 6. The temperatures and the day light conditions at the time of first tillering stage of rice plant, when planted as presenting transplanting practices, are very satisfactory, but the poor day light length, high temperatures and too wet conditions in the time of last-tillering stage(mid or last July) might cause the occurrence of disease such as blast. 7. The heading stage of rice plants at each region through nations when planted as presently practicing method comes when the day light length is short. 8. It was shown that the accumulated average air-temperature at the time of maturing stage was not enough and the heading time was too late, when considered the annual deviations of mean temperatures and low minimum temperatures. 9. The nitrogen content of each plant part at the each growing stage was very high at the stage of vegetative growth when compared with the nitrogen content at the stage of reproductive growth after heading. In this respect it was believed to be important to prevent the nutrient shortages at the reproductive stages, especially after the heading. 10. The area of unsatisfactory irrigation paddy fields and natural rain-fed paddy fields are getting reduced in Korea. The correlation between the rate of reducing unsatisfactory irrigation and natural rain-fed paddy fields and the rate of yield increase were computed. The correlation coefficients(r) between the area of unsatisfactory irrigation paddy fields and yield increase were +0.525, and between the natural rain-fed paddy fields and yield increase, +0.832 and between the unsatisfactory irrigation plus natural rain-fed paddy fields and yield increase, +0.84. And there were. highly significant positive correlations between natural rain-fed paddy fields and yield increases indicating that the less the area of natural rain-fed paddy fields, the greater the yields per unit area. 11. The results obtained from the fertilizer experiments (yield performance trials) conducted in both Korea and Japan showed that the yield of non-fertilized plots per 10 are was 231 kg in Korea and 360 kg in Japan. On the basis of this it might be concluded that the fertility of soil in Korea is lower than that in Japan. Furthermore it was. also found that the yields of non-nitrogen applied plots per 10 are were 236 kg in Korea and 383 kg in Japan. This also indicates that the yields of rice in Korea are largely depending on the nitrogen content in the soil. 12. The followings were obtained when the chemical natures of soils in both Korea and Japan were compared. (1) The content of organic matter, total nitrogen, exchangeable calcium, and magnesium in Korea were no more than the half those in Japan. (2) The content of N/2 chloride and soluble silicate in low-land soil were on the average lower in Korea. (3) The exchange capacity of bases in Korea was no more than half that in Japan. 13. It was also observed by comparing the soil nature of the soil with high yielding capacity with the soil with low yielding capacity that the exchange capacity of bases, exchangeable calcium and magnesium, potassium, phosphorus, manganese, silicate and iron were low in the soil with low yielding capacity. 14. The depth of furrow slice was always deeper in the soil with high yielding capacity, and the depth of furrow slice in Korea was also shallower than that in Japan. 15. Summarizing the various conditions mentioned previously and considering the effects of silicate and trace elements such as manganese and iron besides three elements on the physiological and plant type formation of rice crops, more realistic and more ideal fertilizing practices were proposed. proposed.

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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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