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Mass Production of Gain-of-Function Mutants of Hair Roots in Ginseng (기능획득 돌연변이 인삼 모상근의 대량생산)

  • Ko, Suk-Min;In, Dong-Soo;Chung, Hwa-Jee;Choi, Dong-Woog;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.285-291
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    • 2007
  • This study describes conditions for the mass production of activation-tagged mutant hairy root lines of ginseng by cocultivation with Agrobacterium rhizogenes. Because it is not currently possible to produce progeny from transgenic ginseng, a loss-of-function approach for functional genomics cannot be appliable to this species. A gain-of-function approach is alternatively the choice and hairy root production by cocultivation of A. rhizogenes would be most practical to obtain a large number of mutants. Various sources of explants were subjected to genetic transformation with various strains of A. rhizogenes harboring the activation-tagging vector pKH01 to determine optimum conditions for the highest frequency of hairy root formation on explants. Petiole explants cocultivated with A. rhizogenes R1000 produced hairy roots at a frequency of 85.9% after 4 weeks of culture. Conditions for maximum growth or branching rate of hairy roots were also investigated by using various culture media. Petiole explants cultured on half strength Schenk and Hildebrandt medium produced vigorously growing branched roots at a rate of 2.6 after 4 weeks of culture. A total of 1,989 lines of hairy root mutants were established in this study. These hairy root lines will be useful to determine functions of genes for biosynthesis of ginsenosides.

Influence of medium addition and agitation on the production of embryos in isolated microspore culture of hot pepper (Capsicum annuum L.) (고추의 소포자 배양 시 배지 첨가와 진탕이 배의 생산에 미치는 영향)

  • An, Dong-Ju;Park, Eun-Joon;Kim, Moon-Za
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.30-41
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    • 2011
  • The influences of the agitation as well as the addition of medium during culture on the production of embryos were invested in isolated microspore culture of hot pepper (Capsicum annuum L.). When the culture medium was added during initial liquid culture step of liquid-double layer culture, the embryo yield and quality greatly increased. The most effective time point for medium addition was 5 days after the culture commenced. On the other hand, the effect of medium addition at later double layer culture step in liquid-double layer culture on the embryo production was less compared to that of medium addition during the initial liquid culture step. Agitating the culture for 1 week during later double layer culture step in liquid-double layer culture effectively increased the production of normal cotyledonary embryos. In the case of liquid culture, agitating the culture for 1 week from 7 days after the culture commenced was also effective for embryo development. However, when the total agitation time was longer (2 to 3 weeks) during liquid-double layer culture or liquid culture, the embryos developed abnormally in both cases. The normal cotyledonary embryos obtained in this study successfully developed to plants when transferred to regeneration media. These regenerated plants were either diploid or haploid, and there was a difference in the number of chloroplasts between guard cells of diploid and haploid. These results can be used as an important data for developing an efficient microspore culture system with high quality embryo production in hot pepper.

In vitro shoot propagation of Ranunculus kazusensis Makino, an endangered aquatic plant (멸종위기 수생식물인 매화마름(Ranunculus kazusensis Makino)의 기내 신초 증식)

  • Park, Min Wan;Ryu, Shi Hyun;Nam, Su Hwan;Bae, Kee Hwa
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.325-329
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    • 2017
  • A micropropagation method via callus for Ranunculus kazusensis Makino, an endangered species, was established. When stem segments were cultured on MS media supplemented with 1.0 mg/L IAA, NAA, IBA and 2,4-D, the highest frequency of callus induction was achieved on MS medium supplemented with 1.0 mg/L NAA. Multiple shoot per explant was obtained, the MS medium containing 1.0 mg/L BA and 0.5 mg/L NAA. Additionally, effect of activated charcoal (AC) and sucrose on shoot growth in in vitro culture were examined. The most suitable conditions for shoot growth after 4 weeks of culture were the MS medium with AC and sucrose. This in vitro propagation protocol will be valuable for conservation and mass propagation of this endangered plant.

The change of somatic cell embryogenesis in Kalanchoe pinnata because of agar concentration in stimulating root stress (뿌리 스트레스를 유발하는 agar농도에 따른 Kalanchoe pinnata의 체세포 배 형성 변화)

  • Park, Jongbum;Kim, Jin-Seok;Kim, Donggiun
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.320-324
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    • 2017
  • Development of modern agricultural machinery and accompanying agricultural development cause soil compaction and reduce growth by stressing roots. Kalanchoe pinnata was used to investigate the impact of stress on rooting and changes in plant growth and reproduction. K. pinnata forms somatic embryos capable of asexual reproduction at the edge of leaves. Impact of root pressurization of K. pinnata on somatic embryogenesis and organ differentiation according to external stress factors was investigated by using a high concentration of agar and this phenomenon was studied histologically. Agar concentration in culture media ranged from 0.5%-1.5% to induce a compression effect on roots. The stem and leaf of K. pinnata were subjected to a microtechnique process to study changes in tissue. In vivo, K. pinnata produced 2nd and 3rd plantlets at edges of leaves from lack of water and excessive lighting conditions. In in vitro culture studies, the lower the concentration of agar, the higher the population and the higher the biomass, but plantlet did not occur in leaf bends. Conversely, as concentration of agar increased, increase in the number of individuals was low. Plantlet development occurred only in agar 1.5% medium. The difference in agar concentration was a stressor in the root of K. pinnata, and thus the pattern of asexual reproduction changed from the division method in root to a plantlet generation in leaf. This suggests root pressurization may act as stress and change in the plant reproduction pattern.

Studies on Cultural Characteristics Pestalotiopsis theae causing Leaf Blight on Oriental Persimmon Tree (단감나무 둥근갈색무뉘병원균 Pestalotiopsis theae의 배양적 특성)

  • Chang, Tae-Hyun;Lim, Tae-Heon;Chung, Bong-Koo;Kim, Byung-Sup
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.232-238
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    • 1997
  • Culture conditions affecting mycelial growth and sporulation of P. theae, SP2, SP3 and P. longiseta which causing leaf blight on oriental persimmon leaf blight have been investigated. The optimum temperature for the mycelial growth and sporulation on potato dextrose agar was $25{\sim}30^{\circ}C$ in all the fungi, but was inhibited and finally arrested at 10 and $30^{\circ}C$. The optimum pH for mycelial growth and sporulation were ranged at pH 4.5~5.0 and 5.0~6.0 in all the fungi. Lenonian agar, potato sucrose agar and oatmeal medium were good culture media for the mycelial growth and sporulation of all the fungi. The effective sources of nitrogen and carbon for the mycelial growth were tryptone, glycine, starch, dextrose, galactose and lactose. Glutamic acid, peptone and tryptone were good nitrogen sources for sporulation of the fungi. Sucrose, starch and galactose were also good carbon sources for sporulation. Generally, vitamins had no effect on mycelial growth and sporulation. The pH of the potato dextrose broth inoculated with SP2, SP3 and P. theae and P. longiseta was changed from 7.0 to 4.5~4.7 and 5.0~5.4 after incubating for 10 days, respectively. But, the initial pH of the medium adjusted to 5.0 was lowered to 4.5~4.7 after incubating for 10 days.

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Spore Germination of Some Plant Pathogenic Fungi under Different Soil Conditions in Relation to Soil Fungistasis (토양조건에 따른 몇가지 식물병원균의 포자발아와 토양정균 현상)

  • Lee Min Woong;Choi Hae Jung;Shim Jae Ouk
    • Korean Journal Plant Pathology
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    • v.1 no.3
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    • pp.157-164
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    • 1985
  • Some interactions in various soil conditions, numbers of microbial populations, root rot disease development and rates of spore germiation in three different location of soils were investigated. The calcium and magnesium contents were higher in replanted fields of ginseng (Panax ginseng) at Goesan. Potassium contents were high in replanted field at Poonggi and textural class of the soils was silt loam except for silt clay loam in first cultured field of ginseng at Goesan. For the germination process of Fusarium solani, F. moniliforme, F. oxysporum, and Alternaria panax, the percentage germination of fungal spores was high in double distilled water and Pfeffer's solution as media, whereas the lower rate of germination of spores was observed in soil extracts. Numbers of bacteria were high in replanted field soil at Gumsan, and propagules of fungi in replanted fields at Gumsan and Poonggi were higher than other soils, but higher numbers of actinomycetes were found in the first cultured field of ginseng at Goesan and Poonggi. Fungistasis was induced by higher microbial populations present in soil that was initiated when amended with garlic stalk, crushed bean and ginseng leaves. On the other hand, there was no fungistasis in soil amended with wheat and barley straw, and this tendency was a little difference on the soil sample.

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Application of professor·learning model customized for flipped learning for enhancing basic ability of work - Focused on freshman students in radiology department of specialized colleges (직업기초능력함양을 위한 맞춤식 플립드 러닝 교수·학습모형 적용-전문대학 방사선과 1학년 재학생을 중심으로)

  • Park, Jeongkyu
    • Journal of the Korean Society of Radiology
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    • v.12 no.2
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    • pp.225-231
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    • 2018
  • Recently, new teaching methods for communicating with teachers and students have been emerged according to the trends of decreasing the school-age population and the development of the mass media. We have applied teaching-learning model based on the flip learning to the college students in this work. As a result of the test for the customized flipped learning teaching-learning model in pre-class, the attendance rate of the major subject was 92.3% whereas that in liberal arts courses other than majors revealed 87.6%. This result for attendance rate shows that first year students in the radiology department have been actively participated in pre-class of the major subject than that of the liberal arts curriculum. From comparing the differences between the study group that was applied flipped learning in class and the non-applied group, the research group showed higher scores in knowledge, skills, and attitudes than the comparative group. In addition, more than 90% of the learners improved their responsibility, problem solving ability, creative thinking, cooperative ability, and communication ability through this learning program. From the test for the difference in the role of radiologists in the post class, the mean score was 4.40 for the group applied the teaching-learning model while that for non-applied group was 2.10. Hence, from such results, we see that this teaching-learning model is appropriate and needs to be extended to cultivate basic skills in radiology and relevant vocational education.

Acute Type A Aortic Dissection Mimicking Penetrating Atherosclerotic Ulcer (침투성 동맥경화성 궤양과 흡사한 급성 A형 대동맥 박리증 -1l례 보고 -)

  • Choi, jae-Sung;Kwak, Jae-Gun;Ahn, Hyuk
    • Journal of Chest Surgery
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    • v.36 no.1
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    • pp.30-34
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    • 2003
  • As classical acute aortic dissection, atherosclerotic penetrating ulcers and intramural hematoma have different pathophysiology and natural history, treatment strategy should be different and, therefore, accurate differential diagnosis is necessary. However, these three aortic diseases may be indistinguishable by clinical observation and even by various diagnostic modalities such as cardiac echocardiography, CT and MRI. The patients was a 71-year-old female with chief complaints of anterior chest pain, nausea and vomiting which occurred suddenly 3 days before admission. CT angiographic with 3 dimensional reconstruction shows intramural hematoma in ascending aorta, aortic arch, descending thoracic aorta and right brachiocephalic trunk, heompericardium, and blood in mediastinum and both pleural cavities. The CT angiographic finding of focal out-bulging in the ascending thoracic aorta was diagnosed as penetrating atherosclerotic ulcer. The patient underwent emergency operation under a preoperative diagnosis of penetrating atherosclerotic ulcer with a sign of aortic rupture. In the intraoperative findings, however, intimal tear was seen in the anterior portion of the ascending aorta about 1cm below the brachiocephalic trunk and falselumen appeared after hematoma was removed from the layer of tunica media. We report a case of type A aortic dissection which mimicked clinical and diagnostic features of penetrating atherosclerotic ulcer.

Effects of established cell lines on bovine embryo development during in vitro culture (계대세포를 이용한 소 수정란의 체외배양 체계에 관한 연구)

  • Joo, Suk-chun;Lee, Byeong-chun;Lee, Won-yu;Choi, Yun-seok;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.647-659
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    • 1997
  • To overcome the difficulties of collecting and culture of primary cell from genital tract on embryonic development, the present study was carried out to investigate the critical effect of cell lines, such as BRL and Vero cell and its conditioned medium on the development of early Korean native cattle embryos in vitro. Oocytes collected from slaughterhouse ovaries were matured in TCM199 containing FSH, estradiol-$17{\beta}$ and FBS with granulosa cell monolayer for 24 hours and then fertilized in vitro using frozen-thawed, heparin-treated spermatozoa in TALP for 30 hours. And then early embryos (1-2cell) were cultured in TCM199 containing 10% FBS with BOEC, Granulosa, BRL, Vera cell monolayers and conditioned medium for 2~3 days. Development to morulae and blastocysts were recorded, also examined the number of blastomeres presented a valuable parameter for the evaluation of embryonic development. The early cleavage rates of in vitro fertilized embryos co-cultured, there was no differences between primary cell and cell lines(p<0.05). The rate of development to the later stage, coculture of BRL cell was significantly higher than that of the primary cell(p<0.05). The rates of development to morula and blastocyst were significantly higher in vero cell than BRL, Granulosa, Oviduct epithelial cell conditioned medium. In the result of effect of serum on development of early bovine embryos, the use of media containing serum were significantly higher than the use of not containing one on development of early and later stage of embryos. The result of number of blastomeres in blastocysts, there is no differences between primary cell and cell lines. The blastocysts from coculture were higher than from conditioned medium in blastomere cells. In summary, these experments have proved that the culture system in TCM199 with BRL, Vero cell monolayers is effective on in vitro development of early bovine embryos, In addition, it is effective to development of bovine embryos that containing serum in conditioned medium, or in co-culture rather than in conditioned medium alone. The use of cell lines opponent to primary cells is effective in bovine embryo culture.

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Culture characteristics and genetic relationship of morel mushroom (Morchella spp.) isolates from Korea and other countries (곰보버섯 (Morchella spp.) 수집균주의 배양적특성 및 유전적 유연관계)

  • Min, Gyeong-Jin;Park, Hye-sung;Lee, Eun-ji;Lee, Chan-Jung
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.100-106
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    • 2020
  • Eight morel mushroom species were collected from Korea and other countries. The culture characteristics, genetic relationships, and beta-glucan content of the strains were analyzed. The mycelia of Morchella species exhibited optimal growth when cultured in dark at 25 ℃ in media with pH 7. The mycelia had a distinctive mycelial scent and characteristically changed color, being white initially, and then turning dark yellow to dark brown as it grew. The mycelia were classified into five types based on morphology. The isolates were identified as Morchella conica, two M. sextelata, M. importuna, M. esculenta, and three M. crassipes, based on ITS-rDNA sequences. PCR polymorphisms were variably produced within Morchella spp. using Universal Fungal Fingerprinting Primers (UFPF) and classified into four groups at the intra and inter species level. The strains, KMCC04971 and KMCC04407, showed the same banding pattern as M. conica and M. sextelata, respectively; however, these results were different from those of ITS analysis. Glucan content analysis by strain showed that the KMCC 04973 strain of M. importuna had the highest alpha- and beta-glucan content, at 16.4 g and 33.1 g per 100 g, respectively.