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Studies on Genetics and Breeding in Rainbow Trout(Oncorhynchus mykiss) VII. Fertilization of Fresh Egg with Co-Preserved Sperm and Ultrastructural Changes (무지개 송어의 유전 육종학적 연구 VII. 동결보존시킨 정자와 신선한 난모세포의 수정 및 미세구조적 변화)

  • PARK Hong-Yang;YOON Jong-Man
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.2
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    • pp.79-92
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    • 1992
  • This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

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Herbicidal Phytotoxicity under Adverse Environments and Countermeasures (불량환경하(不良環境下)에서의 제초제(除草劑) 약해(藥害)와 경감기술(輕減技術))

  • Kwon, Y.W.;Hwang, H.S.;Kang, B.H.
    • Korean Journal of Weed Science
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    • v.13 no.4
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    • pp.210-233
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    • 1993
  • The herbicide has become indispensable as much as nitrogen fertilizer in Korean agriculture from 1970 onwards. It is estimated that in 1991 more than 40 herbicides were registered for rice crop and treated to an area 1.41 times the rice acreage ; more than 30 herbicides were registered for field crops and treated to 89% of the crop area ; the treatment acreage of 3 non-selective foliar-applied herbicides reached 2,555 thousand hectares. During the last 25 years herbicides have benefited the Korean farmers substantially in labor, cost and time of farming. Any herbicide which causes crop injury in ordinary uses is not allowed to register in most country. Herbicides, however, can cause crop injury more or less when they are misused, abused or used under adverse environments. The herbicide use more than 100% of crop acreage means an increased probability of which herbicides are used wrong or under adverse situation. This is true as evidenced by that about 25% of farmers have experienced the herbicide caused crop injury more than once during last 10 years on authors' nationwide surveys in 1992 and 1993 ; one-half of the injury incidences were with crop yield loss greater than 10%. Crop injury caused by herbicide had not occurred to a serious extent in the 1960s when the herbicides fewer than 5 were used by farmers to the field less than 12% of total acreage. Farmers ascribed about 53% of the herbicidal injury incidences at their fields to their misuses such as overdose, careless or improper application, off-time application or wrong choice of the herbicide, etc. While 47% of the incidences were mainly due to adverse natural conditions. Such misuses can be reduced to a minimum through enhanced education/extension services for right uses and, although undesirable, increased farmers' experiences of phytotoxicity. The most difficult primary problem arises from lack of countermeasures for farmers to cope with various adverse environmental conditions. At present almost all the herbicides have"Do not use!" instructions on label to avoid crop injury under adverse environments. These "Do not use!" situations Include sandy, highly percolating, or infertile soils, cool water gushing paddy, poorly draining paddy, terraced paddy, too wet or dry soils, days of abnormally cool or high air temperature, etc. Meanwhile, the cultivated lands are under poor conditions : the average organic matter content ranges 2.5 to 2.8% in paddy soil and 2.0 to 2.6% in upland soil ; the canon exchange capacity ranges 8 to 12 m.e. ; approximately 43% of paddy and 56% of upland are of sandy to sandy gravel soil ; only 42% of paddy and 16% of upland fields are on flat land. The present situation would mean that about 40 to 50% of soil applied herbicides are used on the field where the label instructs "Do not use!". Yet no positive effort has been made for 25 years long by government or companies to develop countermeasures. It is a really sophisticated social problem. In the 1960s and 1970s a subside program to incoporate hillside red clayish soil into sandy paddy as well as campaign for increased application of compost to the field had been operating. Yet majority of the sandy soils remains sandy and the program and campaign had been stopped. With regard to this sandy soil problem the authors have developed a method of "split application of a herbicide onto sandy soil field". A model case study has been carried out with success and is introduced with key procedure in this paper. Climate is variable in its nature. Among the climatic components sudden fall or rise in temperature is hardly avoidable for a crop plant. Our spring air temperature fluctuates so much ; for example, the daily mean air temperature of Inchon city varied from 6.31 to $16.81^{\circ}C$ on April 20, early seeding time of crops, within${\times}$2Sd range of 30 year records. Seeding early in season means an increased liability to phytotoxicity, and this will be more evident in direct water-seeding of rice. About 20% of farmers depend on the cold underground-water pumped for rice irrigation. If the well is deep over 70m, the fresh water may be about $10^{\circ}C$ cold. The water should be warmed to about $20^{\circ}C$ before irrigation. This is not so practiced well by farmers. In addition to the forementioned adverse conditions there exist many other aspects to be amended. Among them the worst for liquid spray type herbicides is almost total lacking in proper knowledge of nozzle types and concern with even spray by the administrative, rural extension officers, company and farmers. Even not available in the market are the nozzles and sprayers appropriate for herbicides spray. Most people perceive all the pesticide sprayers same and concern much with the speed and easiness of spray, not with correct spray. There exist many points to be improved to minimize herbicidal phytotoxicity in Korea and many ways to achieve the goal. First of all it is suggested that 1) the present evaluation of a new herbicide at standard and double doses in registration trials is to be an evaluation for standard, double and triple doses to exploit the response slope in making decision for approval and recommendation of different dose for different situation on label, 2) the government is to recognize the facts and nature of the present problem to correct the present misperceptions and to develop an appropriate national program for improvement of soil conditions, spray equipment, extention manpower and services, 3) the researchers are to enhance researches on the countermeasures and 4) the herbicide makers/dealers are to correct their misperceptions and policy for sales, to develop database on the detailed use conditions of consumer one by one and to serve the consumers with direct counsel based on the database.

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A study of compaction ratio and permeability of soil with different water content (축제용흙의 함수비 변화에 의한 다짐율 및 수용계수 변화에 관한 연구)

  • 윤충섭
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.13 no.4
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    • pp.2456-2470
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    • 1971
  • Compaction of soil is very important for construction of soil structures such as highway fills, embankment of reservoir and seadike. With increasing compaction effort, the strength of soil, interor friction and Cohesion increas greatly while the reduction of permerbilityis evident. Factors which may influence compaction effort are moisture content, grain size, grain distribution and other physical properties as well as the variable method of compaction. The moisture content among these parameter is the most important thing. For making the maximum density to a given soil, the comparable optimum water content is required. If there is a slight change in water content when compared with optimum water content, the compaction ratio will decrease and the corresponding mechanical properties will change evidently. The results in this study of soil compaction with different water content are summarized as follows. 1) The maximum dry density increased and corresponding optimum moisture content decreased with increasing of coarse grain size and the compaction curve is steeper than increasing of fine grain size. 2) The maximum dry density is decreased with increasing of the optimum water content and a relationship both parameter becomes rdam-max=2.232-0.02785 $W_0$ But this relstionship will be change to $r_d=ae^{-bw}$ when comparable water content changes. 3) In case of most soils, a dry condition is better than wet condition to give a compactive effort, but the latter condition is only preferable when the liquid limit of soil exceeds 50 percent. 4) The compaction ratio of cohesive soil is greeter than cohesionless soil even the amount of coarse grain sizes are same. 5) The relationship between the maximum dry density and porosity is as rdmax=2,186-0.872e, but it changes to $r_d=ae^{be}$ when water content vary from optimum water content. 6) The void ratio is increased with increasing of optimum water content as n=15.85+1.075 w, but therelation becames $n=ae^{bw}$ if there is a variation in water content. 7) The increament of permeabilty is high when the soil is a high plasticity or coarse. 8) The coefficient of permeability of soil compacted in wet condition is lower than the soil compacted in dry condition. 9) Cohesive soil has higher permeability than cohesionless soil even the amount of coarse particles are same. 10) In generall, the soil which has high optimum water content has lower coefficient of permeability than low optimum water content. 11) The coefficient of permeability has a certain relations with density, gradation and void ratio and it increase with increasing of saturation degree.

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A Study on the Effect of Water Soluble Extractive upon Physical Properties of Wood (수용성(水溶性) 추출물(抽出物)이 목재(木材)의 물리적(物理的) 성질(性質)에 미치는 영향(影響))

  • Shim, Chong-Supp
    • Journal of the Korean Wood Science and Technology
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    • v.10 no.3
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    • pp.13-44
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    • 1982
  • 1. Since long time ago, it has been talked about that soaking wood into water for a long time would be profitable for the decreasing of defects such as checking, cupping and bow due to the undue-shrinking and swelling. There are, however, no any actual data providing this fact definitly, although there are some guesses that water soluble extractives might effect on this problem. On the other hand, this is a few work which has been done about the effect of water soluble extractives upon the some physical properties of wood and that it might be related to the above mentioned problem. If man does account for that whether soaking wood into water for a long time would be profitable for the decreasing of defects due to the undue-shrinking and swelling in comparison with unsoaking wood or not, it may bring a great contribution on the reasonable uses of wood. To account for the effect of water soluble extractives upon physical properties of wood, this study has been made at the wood technology laboratory, School of Forestry, Yale university, under competent guidance of Dr. F. F. Wangaard, with the following three different species which had been provided at the same laboratory. 1. Pinus strobus 2. Quercus borealis 3. Hymenaea courbaril 2. The physical properties investigated in this study are as follows. a. Equilibrium moisture content at different relative humidity conditions. b. Shrinkage value from gre condition to different relative humidity conditions and oven dry condition. c. Swelling value from oven dry condition to different relative humidity conditions. d. Specific gravity 3. In order to investigate the effect of water soluble extractives upon physical properties of wood, the experiment has been carried out with two differently treated specimens, that is, one has been treated into water and the other into sugar solution, and with controlled specimens. 4. The quantity of water soluble extractives of each species and the group of chemical compounds in the extracted liquid from each species have shown in Table 36. Between species, there is some difference in quantity of extractives and group of chemical compounds. 5. In the case of equilibrium moisture contents at different relative humidity condition, (a) Except the desorption case at 80% R. H. C. (Relative Humidity Condition), there is a definite line between untreated specimens and treated specimens that is, untreated specimens hold water more than treated specimens at the same R.H.C. (b) The specimens treated into sugar solution have shown almost the same tendency in results compared with the untreated specimens. (c) Between species, there is no any definite relation in equilibrium moisture content each other, however E. M. C. in heartwood of pine is lesser than in sapwood. This might cause from the difference of wood anatomical structure. 6. In the case of shrinkage, (a) The shrinkage value of the treated specimen into water is more than that of the untreated specimens, except anyone case of heartwood of pine at 80% R. H. C. (b) The shrinkage value of treated specimens in the sugar solution is less than that of the others and has almost the same tendency to the untreated specimens. It would mean that the penetration of some sugar into the wood can decrease the shrinkage value of wood. (c) Between species, the shrinkage value of heartwood of pine is less than sapwood of the same, shrinkage value of oak is the largest, Hymenaea is lesser than oak and more than pine. (d) Directional difference of shrinkage value through all species can also see as other all kind of species previously tested. (e) There is a definite relation in between the difference of shrinkage value of treated and untreated specimens and amount of extractives, that is, increasing extractives gives increasing the difference of shrinkage value between treated and untreated specimens. 7. In the case of swelling, (a) The swelling value of treated specimens is greater than that of the untreated specimens through all cases. (b) In comparison with the tangential direction and radial direction, the swelling value of tangential direction is larger than that of radial direction in the same species. (c) Between species, the largest one in swelling values is oak and the smallest pine heartwood, there are also a tendency that species which shrink more swell also more and, on the contrary, species which shrink lesser swell also lesser than the others. 8. In the case of specific gravity, (a) The specific gravity of the treated specimens is larger than that of untreated specimens. This reversed value between treated and untreated specimens has been resulted from the volume of specimen of oven dry condition. (b) Between species, there are differences, that is, the specific gravity of Hymenaea is the largest one and the sapwood of pine is the smallest. 9. Through this investigation, it has been concluded that soaking wood into plain water before use without any special consideration may bring more hastful results than unsoaking for use of wood. However soaking wood into the some specially provided solutions such as salt water or inorganic matter may be dissolved in it, can be profitable for the decreasing shrinkage and swelling, checking, shaking and bow etc. if soaking wood into plain water might bring the decreasing defects, it might come from even shrinking and swelling through all dimension.

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The Variation of Natural Population of Pinus densiflora S. et Z. in Korea (III) -Genetic Variation of the Progeny Originated from Mt. Chu-wang, An-Myon Island and Mt. O-Dae Populations- (소나무 천연집단(天然集團)의 변이(變異)에 관(關)한 연구(硏究)(III) -주왕산(周王山), 안면도(安眠島), 오대산(五臺山) 소나무집단(集團)의 차대(次代)의 유전변이(遺傳變異)-)

  • Yim, Kyong Bin;Kwon, Ki Won
    • Journal of Korean Society of Forest Science
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    • v.32 no.1
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    • pp.36-63
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    • 1976
  • The purpose of this study is to elucidate the genetic variation of the natural forest of Pinus densiflora. Three natural populations of the species, which are considered to be superior quality phenotypically, were selected. The locations and conditions of the populations are shown in table 1 and 2. The morphological traits of tree and needle and some other characteristics were presented already in our first report of this series in which population and family differences according to observed characteristics were statistically analyzed. Twenty trees were sampled from each populations, i.e., 60 trees in total. During the autumn of 1974, matured cones were collected from each tree and open-pollinated seeds were extracted in laboratory. Immediately after cone collection, in closed condition, the morphological characteristics were measured. Seed and seed-wing dimensions were also studied. In the spring of 1975, the seeds were sown in the experimental tree nursery located in Suweon. And in the April of 1976, the 1-0 seedlings were transplanted according to the predetermined experimental design, randomized block design with three replications. Because of cone setting condition. the number of family from which progenies were raised by populations were not equal. The numbers of family were 20 in population 1. 18 in population 2 and 15 in population 3. Then, each randomized block contained seedlings of 53 families from 3 populations. The present paper is mainly concerned with the variation of some characteristics of cone, seed, needle, growth performance of seedlings, and chlorophyll and monoterpene compositions of needles. The results obtained are summerized as follows. 1. The meteorological data obtained by averaging the records of 30 year period, observed from the nearest station to each location of populations, are shown in Fig. 3, 4, and 5. The distributional pattern of monthly precipitation are quite similar among locations. However, the precipitation density on population 2, Seosan area, during growing season is lower as compared to the other two populations. Population 1. Cheong-song area, and population 3, Pyong-chang area, are located in inland, but population 2 in the western seacoast. The differences on the average monthly air temperatures and the average monthly lowest temperatures among populations can hardly be found. 2. Available information on the each mother trees (families) studied, such as age, stem height, diameter at breast height, clear-bole-length, crown conditions and others are shown in table 6,7, and 8. 3. The measurements of fresh cone weight, length and the widest diameter of cone are given in Tab]e 9. All these traits arc concerned with the highly significant population differences and family differences within population. And the population difference was also found in the cone-index, that is, length-diameter ratio. 4. Seed-wing length and seed-wing width showed the population differences, and the family differences were also found in both characteristics. Not discussed in this paper, however, seed-wing colours and their shapes indicate the specificity which is inherent to individual trees as shown in photo 3 on page 50. The colour and shape are fully the expression of genetic make up of mother tree. The little variations on these traits are resulted from this reason. The significant differences among populations and among families were found in those characteristics, such as 1000-seed weight, seed length, seed width, and seed thickness as shown in table 11. As to all these dimensions, the values arc always larger in population 1 which is younger in age than that of the other two. The population differences evaluated by cone, seed and seed-wing sizes could partly be attributed to the growth vigorousity. 5. The values of correlation between the characteristics of cone and seed are presented in table 12. As shown, the positive correlations between cone diameter and seed-wing width were calculated in all populations studied. The correlation between seed-wing length and seed length was significantly positive in population 1 and 3 but not in population 2, that is, the r-value is so small as 0.002. in the latter. The correlation between cone length and seed-wing length was highly significant in population 1, but not in population 2. 6. Differences among progenies in growth performances, such as 1-0 and 1-1 seedling height and root collar diameter were highly singificant among populations as well as families within population(Table 13.) 7. The heritability values in narrow sense of population characteristics were estimated on the basis of variance components. The values based on seedling height at each age stage of 1-1 and 1-0 ranged from 0.146 to 0.288 and the values of root collar diameter from 0.060 to 0.130. (Table 14). These heritability values varied according to characteristics and seedling ages. Here what must be stated is that, for calculation of heritability values, the variance values of population was divided by the variance value of environment (error) and family and population. The present authors want to add the heritability values based on family level in the coming report. It might be considered that if the tree age is increased in furture, the heritability value is supposed to be altered or lowered. Examining the heritability values studied previously by many authors, in pine group at age of 7 to 15, the values of height growth ranged from 0.2 to 0.4 in general. The values we obtained are further below than these. 8. The correlation between seedling growth and seed characteristics were examined and the values resulted are shown in table 16. Contrary to our hypothetical premise of positive correlation between 1-0 seedling height and seed weight, non-significance on it was found. However, 1-0 seedling height correlated positively with seed length. And significant correlations between 1-0 and 1-1 seedling height are calculated. 9. The numbers of stomata row calculated separately by abaxial and adaxial side showed highly significant differences among populations, but not in serration density. On serration density, the differences among families within population were highly significant. (Table 17) A fact must be noted is that the correlation between stomata row on abaxial side and adaxial side was highly significant in all populations. Non-significances of correlation coefficient between progenies and parents regarding to stomata row on abaxial side were shown in all populations studied.(Table 18). 10. The contents of chhlorophyll b of the needle were a little more than that of chlorophyll a irrespective of the populations examined. The differences of chlorophyll a, b and a plus b contents were highly significant but not among families within populations as shown in table 20. The contents of chlorophyll a and b are presented by individual trees of each populations in table 21. 11. The occurrence of monoterpene components was examined by gas liquid chromatography (Shimazu, GC-1C type) to evaluate the population difference. There are some papers reporting the chemical geography of pines basing upon monoterpene composition. The number of populations studied here is not enough to state this problem. The kinds of monoterpene observed in needle were ${\alpha}$-pinene, camphene, ${\beta}$-pinene, myrcene, limonene, ${\beta}$-phellandrene and terpinolene plus two unknowns. In analysis of monoterpene composition, the number of sample trees varied with population, I.e., 18 families for population 1, 15 for population 2 and 11 for population3. (Table 22, 23 and 24). The histograms(Fig. 6) of 7 components of monoterpene by population show noticeably higher percentages of ${\alpha}$-pinene irrespective of population and ${\beta}$-phellandrene in the next order. The minor Pinus densiflora monoterpene composition of camphene, myrcene, limonene and terpinolene made up less than 10 percent of the portion in general. The average coefficients of variation of ${\alpha}$-pinene and ${\beta}$-phellandrene were 11 percent. On the contrary to this, the average coefficients of variation of camphene, limonene and terpinolene varied from 20 to 30 percent. And the significant differences between populaiton were observed only in myrcene and ${\beta}$-phellandrene. (Table 25).

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