• Title/Summary/Keyword: Infertile

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A Study on the Induction of Infertility of Largemouth Bass (Micropterus salmoides) by CRISPR/Cas9 System (CRISPR/Cas9 System을 활용한 배스의 불임 유도에 대한 연구)

  • Park, Seung-Chul;Kim, Jong Hyun;Lee, Yoon Jeong
    • Korean Journal of Environment and Ecology
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    • v.35 no.5
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    • pp.503-524
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    • 2021
  • A largemouth bass (Micropterus salmoides) is an ecosystem disturbance fish species at the highest rank in the aquatic ecosystem, causing a serious imbalance in freshwater ecosystems. Although various attempts have been made to eradicate and control largemouth bass, no effective measures were found. Therefore, it is necessary to find an approach to maximize the effective population reduction based on the unique characteristics of largemouth bass. This study used the transcriptome analysis to derive 182,887 unigene contigs and select 12 types of final target sequences for applying the CRISPR/Cas9 system in the genes of IZUMO1 and Zona pellucida sperm-binding protein, which are proteins involved in sperm-egg recognition. After synthesizing 12 types of sgRNA capable of recognizing each target sequence, 12 types of Cas9-sgRNA ribonucleoprotein (RNP) complexes to be used in subsequent studies were prepared. This study searched the protein-coding gene of sperm-egg through the Next Generation Sequencing (NGS) and edited genes through the CRISPR/Cas9 system to induce infertile individuals that produced reproductive cells but could not form fertilized eggs. Through such a series of processes, it successfully established a composition development process for largemouth bass. It is judged that this study contributed to securing the valuable basic data for follow-up studies to verify its effect for the management of ecological disturbances without affecting the habitat of other endemic species in the same water system with the largemouth bass.

Production of Transgenic Murine Embryos using Haploid Spermatids Transfected with EGFP Gene (EGFP 유전자가 도입된 반수체 정자세포에 의한 형질전환 설치류 난자의 생산)

  • Kang, K.Y.;Song, S.J.;Lee, H.T.;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.305-315
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    • 2001
  • In this study, the production of transgenic embryo was attempted by microinjection or round spermatid cultured with foreign DNA. At first, the expression of haploid spermatids specific gene, mTP1 in mouse and hPrm2 in hamster spermatids were investigated by RT-PCR method in testes of young mice and hamster testis. The specific gene expression first appeared at 18 days post partum (dpp) in mice spermatid and 20 dpp in hamster spermatid. Therefore, the round spermatids isolated from 17 dpp mice and 19 dpp hamster were used for the introduction of foreign EGFP gene into haploid round spermatids. For the introduction of EGFP gene haploid round spermatids suspended in medium including EGFP gene were treated with a different electric field strength at 0.11, 0.18 and 0.44 ㎸/cm. After electrical stimulation, viability of testicular sperm cells and 67.6%, 66.4% and 49.9%, in mice and 62.6%, 57.9% and 27% in hamster, respectively. These values were significantly lower than those of non-treated control groups 80.5% in mouse and 69.1% in hamster After 72 hrs culture, the highest expression rate of EGFP gene, 28.5% in mice and 32.1% in hamster were obtained from tile spermatogenic cells electroporated by the field strength or 0.18 ㎸/cm. Then, the ability of fertilization and embryonic development of haploid spermatids transfected with foreign EGFP gene were estimated by the microinjection of spermatids into hamster oocytes. The Irate pronuclear formation rate (77.5%) was lower than non-treated control (80%), and the cleavage rate of the treated group (58.8%) was lower than control (65%). To prove the foreign EGFP integration in hamster embryos, 2-cell stage hamster embryos were subjected to the observation under the fluorescence microscope, and the PCR analysis. As a result, about 44% of 2-cell embryos were showed the integration of EFGP gene into their genome. Therefore, These results suggest the possibility to produce transgenic hamsters by microinjection of haploid spermatid transfected with foreign DNA.

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The effect of potash on the growth and yields of soybean at different level of soil fertility and application of fertilizer (토양(土壤)과 시비(施肥)를 달리할때 대두생육(大豆生育)에 미치는 가리(加里)의 영향(影響))

  • Cho, C.Y.;Maeng, D.W.
    • Applied Biological Chemistry
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    • v.10
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    • pp.107-112
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    • 1968
  • In order to study the effect of potash on the growth and yields of Soybean at different level of soil fertility and application of fertilizer (nitrogen, phosphate and calcium), $2^3$factorical experiment was carried out by pat culture with variety 'Chang-dan-baec-muc' which is most spreaded variety in Korea. The experiment consisted of five replications in a randomized block experiment with three factors (soil, fertilization and potash). Treatment were at two levels; infertile and fertile soil, none and some of fertilization and potash. Thus, the experiment comprised eight treatment combinations which consisted of all combinations. The results of this experiment are as follows: 1. No effect of each of three factors on flowering date was found. 2. Leaf-yellowing and maturing date was quickened on the fertile soil but no effect of fertilization and potash was found. 3. More premature leaf-yellowing was found on the fertile soil. 4. Deeper leaf colour cuss showed on the fertile soil and in the case of fertilization but no effect of potash was found. 5. Increasing tendency of following character: length and width of leaf, height and dia of stem, number of branches and pods; was most remarkable on the fertile soil. Application of fertilizer showed also remarkable tendency of increasing, while increasing tendency of potash was the least. 6. Same tendency was found with following charactors; weight of total plant. stem and shell, and commercial grains, weight of 100 grain and number of commercial grains. 7. As the results of analysis of variance for weight of commercial grain it, was found the teach of the three factors increased soybean yields significantly (weight of commercial grain) but the effect of potash was less than the other two factors. No significant interaction was found among three factors. 8. Greater effect of potash on increasing soybean yields was found on the fertile soil, and in the case of fertilization.

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A Study of the Value of Contemporary Urban Agriculture as Represented by the Saekgyeong(穡經) by Seogye Park Se-dang(西溪 朴世堂) (서계 박세당(西溪 朴世堂) 색경(穡經)에 표방된 현대 도시농업적 가치에 관한 연구)

  • Lim, Jung-Eon;Sung, Jong-Sang
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.33 no.1
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    • pp.76-87
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    • 2015
  • The present study examines the Saekgyeong (Classic of Husbandry; 1676), an agricultural manual dating from the Joseon Dynasty (1392-1910), and the agricultural thought of its author by Park Se-dang (pen name: Seogye; 1629-1703), a scholar. Its purpose lies in exploring the value of contemporary urban agriculture based on an examination of the attitudes toward agriculture, the values pursued through agriculture, and the ways of dealing with and using land as evinced by the classic and its author. Confirmed through an examination of Park's agricultural philosophy and the Saekgyeong, the results of the present study are as follow. First, there is the socioeconomic value of pursuing the stability of and promoting the economic independence of indigent petty peasants through productivity improvement. Second, there is the experiential value of exploration through experience and agricultural field practice for study. Third, there is the environmental value of endeavoring to overcome an infertile natural environment through agricultural methods that sought to accommodate the land by reading the flow and phenomena of nature. Fourth, there is the practical value of compiling the Saekgyeong and seeking to broaden its use as a guidebook containing agricultural methods appropriate to the land and the wisdom for life.8) When examined in terms of contemporary urban agriculture, the significance of the four values above is as follows: the socioeconomic effect of encouraging urban agricultural activities as a means of welfare for socially alienated classes and promoting the creation of jobs; the enhancement of the significance of study through hands-on activities from an educational perspective; the recycling and recovery of resources and the enhancement of environmental consciousness for the recovery of urban ecology; and a practical spirit that seeks to contribute directly to society through academic research that contributes to practical life and approaches familiar to the populace. The present study sought to find the value of urban agriculture, under discussion in diverse ways in recent years, in the thoughts of our ancestors, who pondered on agriculture. Despite differences in the periodic background, the significance of the present study lies in its in-depth reexamination of the fundamental significance of diverse agricultural values that are being pursued today.

Effect and Optimum Quantities of N and K for Chinese Cabbage (배추에 대(對)한 N-K의 효과 및 적량(適量)에 관(關)한 연구(硏究))

  • Oh, Wang-Kun;Han, Sang-Kyung;Kim, Seong-Bae
    • Korean Journal of Soil Science and Fertilizer
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    • v.14 no.4
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    • pp.219-223
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    • 1982
  • The results of study on the yield and absorption of N and K applied to the chinese cabbage (Brassica pekinensis) which was grown at comparatively infertile red-yellow soil in the fall of 1980 are summerized as below. 1. The optimum quantities of potash to be applied for the autumn chinese cabbage is suggested in the range of 15kg-20kg/10a, $K_2O$. 2. Combined treatments of N and K showed a positive interaction by promoting the effect of the other element. The increase of Nitrogen absorption from the additionally applied 10kgs of nitrogen on top of 15kg N/10a showed only 16 per cent at potash minus plot, while that of potash applied plot showed 60 per cent. The cabbage yield increase therefor, at high rate of nitrogen only was 1,700kgs per 10 are over low rate of nitrogen, while high nitrogen plus potash treatment produced additional 2,200kgs over the low rates of N and K. 3. Additional 10kgs of nitrogen applied on top of 15kgs N/10a resulted in increasing soil potassium uptake; 4kgs of soil potassium was additionally absorbed at potash applied plot and 6.5kgs at potash minus plot. 4. Utilization of applied potash was greater at low rates of N, and K application (each 15kgs of N and $K_2O$ per 10a) where 77 per cent of applied potash was utilized, while high rates of N and K application (each 25kgs of N and $K_2O$ per 10a) showed 44 per cent of potash utilization rate. Lower utilization of applied potash at high rate of nitrogen application is due probably to greater uptake of soil potassium. 5. N and K contents in the chinese cabbage are 0.17-0.20% and 0.35-0.43% respectively and these were not statistically correlated with the yields.

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The Role of the Endometrium and Embryo in Human Implantation (인간 착상 과정에 자궁내막과 배아의 역할)

  • Jee, Byung-Chul
    • Development and Reproduction
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    • v.13 no.1
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    • pp.1-11
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    • 2009
  • Implantation itself is governed by an array of endocrine, paracrine and autocrine modulators, of embryonic and maternal origin. Window of implantation is the unique temporal and spatial expression of factors allows the embryo to implant via signaling, appositioning, attachment, and invasion in a specific time frame of $2{\sim}4$ days. When the embryo has arrived in the uterine cavity, a preprogrammed sequence of events occurs, which involves the production and secretion of a multitude of biochemical factors such as cytokines, growth factors, and adhesion molecules by the endometrium and the embryo, thus leading to the formation of a receptive endometrium. Cytokines such as LIF, CSF-1, and IL-1 have all been shown to play important roles in the cascade of events that leads to implantation. Integrin, L-selectin ligands, glycodelin, mucin-1, HB-EGF and pinopodes are involved in appositioning and attachment. The embryo also produces cytokines and growth factors (ILs, VEGF) and receptors for endometrial signals such as LIF, CSF-1, IGF and HB-EGF. The immune system and angiogenesis play an important role. The usefulness of these factors to assess endometrial receptivity and to estimate the prognosis for pregnancy in natural and artificial cycles remains to be proven. Integrins, pinopodes, glycodelin and LIF (from biopsies) are promising candidates; from uterine flushings, glycodelin and LIF are also candidates. The ideal serum marker is not available, but VEGF, glycodelin and CSF have some clinical implications. Further evaluation that includes larger groups of infertile women and fertile controls are needed to elucidate whether their presence in plasma, flushing fluid, or endometrial samples can be used as some kind of a screening tool to assess endometrial function and prognosis for pregnancy before and after artificial reproductive therapy. A better understanding of their function in human implantation may lead to therapeutic intervention, thereby improving the success rate in reproduction treatment. New molecular techniques are becoming available for measuring both embryonic and endometrial changes prior to and during implantation. The use of predictive sets of markers may prove to be more reliable than a single marker. Ultimately, the aim is to use these tools to increase implantation in artificial cycles and consequently improve live-birth rates.

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Clomiphene Citrate on Male Infertility (남성 불임증 환자에 대한 Clomiphene의 효과)

  • Lee, Kang-Hyun;Lee, Hee-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.8 no.2
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    • pp.45-55
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    • 1981
  • Clomiphene citrate. antiestrogen, was given to 39 infertile males whose spermatogenesis were disturbed and the efficacy of the drug was evaluated at the Department of Urology in 1980. (Table 1). Patients were divided into 3 clinical observation groups such as group I composed of 19 cases of idiopathic azoospermia, group II consisted of 15 cases of oligospermia following the vasovasostomy, and group III comprised 5 cases of testicular azoospermia. (Table 2). Clinical characteristics of these patients were as follows: Age of the patients ranged from 26 to 43 years old with mean of 34, and that of their wives ranged from 24 to 41 years old with mean of 31. Duration of marital life ranged from 1 to 21 years with mean of 5 years. Sizes of testis ranged from 6 to 25 ml with mean of 16 ml. Coital frequency ranged from 0.5 to 6 per week with mean of 2.4 per week. Levels of plasma FSH ranged from 3.15 to 23.06 lU/1 with mean of 8.15 lU/1, those of LH ranged from 2.98 to 19.89 lU/1 with mean of 8.18 lU/1 and those of testosterone ranged from 3.09 to 9.97 ng/ml with mean of 6.48 ng/ml. (Table 3). Clomiphene citrate was given in dosage of 50 mg per day (in d.) orally to 31 patients for 3 to 9 months and in dosage of 100 mg per day (b.i.d.) orally to 8 patients for 3 to 9 months. (Table 8). Semen samples were analysed monthly on each patient by routine analysis techniques. For the assessment of the efficacy of Clomiphene citrate on faulty spermatogenesis following empirical criteria were used: For semen quality: Improvement (I) represents that semen parameter increased more than 25% from basal level after the treatment, Unchange (U) expresses that semen parameter increased less than 25% of basal level or not changed after the treatment and Deterioration (D) means that semen parameter decreased from basal level after the treatment. For fertility unit (total counts ${\times}$ motility ${\times}$ morphology ${\div}10^6$): Improvement (I) represents that fertility unit increased more than 10 units after the treatment, Unchange (U) expresses that fertility unit increased less than 10 units or not changed after the treatment, and Deterioration (D) means that fertility unit decreased after the treatment. (Table 4). Results obtained from the Clomiphene therapy were as follows: Changes of spermiograme before and after the Oomiphene therapy shown in the Table 5. Sperm counts increased from 23 to 31 ${\times}10^6$/ml in group I, from 17 to 29 ${\times}10^6$/ml in group II. Other parameters of spermiogramme were not changed significantly after the treatment. Fertility units increased from 14 to 18 units after the treatment in group I, and from 16 to 18 units after the treatment in group II. Effectiveness of Clomiphene citrate on spermatogenesis was summarised in the Tables 6 and 7. After the treatment, sperm count increased in 11 patients, motility increased in 6 patients, morphology increased in 4 patients and fertility units increased in 9 patients. No sperm could be produced by Clomiphene citrate in group III of testicular azoospermia. Dosage of 50 mg of Clomiphene citrate per day for 3 to 6 months was proved to be the most effective in the present series. (Table 8). Pregnancy occurred in 2 patients after the treatment. No particular side effects were noted by the treatment. Pharmacologic compounds used for male infertility were shown in the Table 9. Reported results of Clomiphene citrate were shown in the Table 10.

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Studies on the Development of Acid Tolerant and Superior Nitrogen Fixation Symbionts for Pasture on Hilly Land -II. Selection of Acid Tolerant R. meliloti in virto and Inoculation Effect in Soils (야산(野山) 목초지용(牧草地用) 내산성(耐酸性) 우수(優秀) 질소고정균주(窒素固定菌株) 개발(開發) -II. 내산성(耐酸性) R. meliloti 의 기내선발(器內選拔) 및 토양(土壤) 접종효과(接種效果))

  • Kang, Ui-Gum;Choi, Ju-Hyeon;Cho, Kang-Jin;Jung, Yeun-Tae;Cho, Moo-Je
    • Korean Journal of Soil Science and Fertilizer
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    • v.22 no.1
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    • pp.72-77
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    • 1989
  • A study was conducted to obtain acid tolerant and superior Rhizobium meliloti strain for alfalfa on hilly acid soils. With host plant, eight isolates of R, meliloti selected in the vicinity of Milyang were evaluated for their ability to establish symbiotic effectiveness in acidified tube culture medium and vermiculite pot with different urea levels. Among isolates "YA03" was characterized for the ability to manifest to acid tolerance in three different soils of which pH were 5.0, 6.0, and 7.5. The results obtained were summarized as follows: 1. Of eight isolates "YA03" and "YA09" performed nodule in the tube medium of pH5.0, and in the symbiotic effectiveness YA03 was superior to others. 2. Alfalfa growth and $N_2-fixing$ activity by the inoculation of "YA03" isolate were better than others at the level of urea 1.25mM as well as nonurea. 3. Application of urea with inoculation of "YA03" islate to alfalfa was one of the effective factors for symbiotic effectiveness. 4. In infertile soil of pH5.0 inoculation of R. meliloti "YA03" to alfalfa caused the increase of shoot dry matter of 320% as compared to the control, and the total amount of yield was 131% as much yield in moderate fertile soil of pH7.5. Finally R. meliloti "YA03" isolate was selected as an acid tolerant strain.

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Comparison of Anti-thrombosis, Antioxidant and Anti-diabetes Activities amongDifferent Cultivars of Hibiscus cannabinus L. Seeds (케나프 품종별 종자 추출물의 항혈전, 항산화, 항당뇨 활성의 비교)

  • Lee, Yun-Jin;Kang, Deok-Gyeong;Kim, Ji-Min;Kim, Young-Min;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.32 no.8
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    • pp.633-640
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    • 2022
  • Recently, kenaf (Hibiscus cannabinus L.) has been highlighted due to its rapid growth in dry, infertile land and its versatile applications in textiles, paper, feed, food, medicine, and phyto-remediation. In Korea, a new cultivar, 'Jangdae', which has the merit of feasible seed-gathering, was established in 2013. However, most of the kenaf seeds used are still imported, and a comparison study of useful bio-activities between Jangdae and imported seeds has not been conducted. In this study, anti-thrombosis, antioxidant and anti-diabetic activities of the ethanol extracts of the Jangdae seed (E-JS) and 8 imported cultivar seeds (E-ICS) were investigated. The E-JS has 14.4 mg/g of total polyphenol and 5.3 mg/g of total flavonoids, whereas the E-ICS has 5.9~8.7 mg/g and 0.7~1.5 mg/g, respectively. Among the 9 kenaf seed extracts, IT 181215 and IT 207877 seed extracts showed 1.92 and 1.88-folds extended thrombin time (TT), and IT 207886 seed extracts showed 3.04-folds extended activated partial thromboplastin time (aPTT). All extracts, except Hong-ma seed, showed 1.36~1.56-folds extended prothrombin time (PT). The E-JS showed strong anti-thrombosis activities with 1.46-folds (TT), 1.38-folds (PT) and 2.37-folds (aPTT) compared to those of solvent controls. The E-JS has the strongest antioxidant activities among the seed extracts, with 0.48 mg/ml of RC50 for ABTS cation and 0.27 mg/ml of RC50 for nitrite scavenging. Meanwhile, the E-JS and IT 181215 extracts showed strong inhibition against α-glucosidase with 10.3~10.6% inhibition at 0.5 mg/ml. Our results suggest the E-JS has potential as an anti-thrombosis, antioxidant and anti-diabetic agent.

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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