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Bacteriological Quality of Sea Water in Deukryang Bay, Korea (득량만 해수의 세균학적 수질)

  • CHANG Dong-Suck;JEONG Eun-Tak;YU Hong-Sik;LEE Eun-Woo;LIM Sung-Mee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.1
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    • pp.77-81
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    • 1998
  • A bacteriological study of sea water in Deukryang Bay was conducted to evaluate sanitary conditions of the bay and compliance of waters with the recommended bacteriological criteria for the designated area of shellfish cultivation. Sea water samples were collected at the established sampling stations (fig. 1) from May 1995 to November 1996. During the study period, coliform group, fecal coliform, classification of coliform group with IMViC reactions and pathogenic vibrios were analyzed. Coliform group and feral coliform MPN's were ranged from $<\;3.0\~4600/100m{\ell}\;and\;<\;30\~1,100/100m{\ell}$, respectively. The bacteriological criteria of sea water in shellfish growing area should be less than 70 per l00ml of sea water for median value of coliform MPN, and below $10\%$ of the samples which contain over than 230 for coliform MPN or over than 43 for fecal coliform MPN, Most of the waters from 26 sampling stations were complied water coliform criteria recommended for designated shellfish growing area. Then, the ratios of the samples with move than 230/10ml of coliform group MPN and more than 43/100ml of fecal coliform MPN were $7.4\%$ and $8.5\%$, respectively. The bacterial density of the sea water was deeply affected by rainfall amount. For example, coliform bacterial counts of sea watery after 48 hours from 93 mm rainfall were $6\~7$ times higher than those of without rainfall. During the study period, infectious bacteria such as Vibrio cholerae, Salmonella sp. an d Shigella sp. were not detected from the samples, but detection ratios of Vibrio parahaemolyticus and Vibrio vulnificus were $15\~20\%$ in summer months.

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Increased Production of an Alkaline Protease from Bacillus clausii I-52 by Chromosomal Integration (Bacillus clausii I-52의 Chromosomal Integration에 의한 Alkaline Protease의 생산성 향상)

  • Joo, Han-Seung;Park, Dong-Chul;Choi, Jang-Won
    • Journal of agriculture & life science
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    • v.46 no.1
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    • pp.163-176
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    • 2012
  • TTo increase productivity of a strong extracellular alkaline protease (BCAP), stable strains of Bacillus clausii I-52 carrying another copy of BCAP gene in the chromosome were developed. Integrative vector, pHPS9-fuBCAP carrying BCAP promoter, ribosome binding site, signal sequence and active protease gene was constructed and transferred into B. clausii I-52, and integration of the constructed plasmid into chromosome was identified by PCR. An investigation was carried out on BCAP production by B. clausii I-52 and transformant C5 showing the highest relative activity of alkaline protease using submerged fermentation. Maximum enzyme activity was produced when cells were grown under the submerged fermentation conditions at $37^{\circ}C$ for 48 h with an aeration rate of 1 vvm and agitation rate of 650 rpm in a optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_47H_2O$ 0.01%, $FeSO_47H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). A protease yield of approximately 134,670U/ml was achieved using an optimized media, which show an increase of approximately 1.6-fold compared to that of non-transformant (83,960 U/ml). When the stability of transformant C5 was examined, the integrated plasmid pHPS9-fuBCAP was detected in the transformant after cultivation for 8 days, suggesting that it maintained stably in the chromosomal DNA of transformant C5.

Multiple Shoot Induction on the New Cltivar, Saxifraga fortunei 'Greenstar' by Different Media and Plant Growth Rregulators (배지 및 생장조절제 종류를 이용한 바위떡풀 신품종 '그린스타' 의 다신초 유도)

  • Suh, Jong-Taek;Ryu, Seung-Yeol;Yoo, Dong-Lim;Nam, Chun-Woo;Hur, Youn-Young
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.2
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    • pp.83-86
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    • 2010
  • This study was conducted to develop in vitro propagation techniques of new cultivar, 'Greenstar', bred by Highland Agriculture Research Center. The multiple shoot induction and plant growth of in vitro plant were analyzed by MS media concentration (1/2 MS, 1 MS and 2 MS medium), plant growth regulators and its proper concentration; CPPU [Forchlorfenuron(N-(2-chloro-4-pridyl)-3-phenylurea) (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), thidiazuron [(TDZ), (0.01, 0.1, 0.5 and $1.0mg{\cdot}L^{-1}$), zeatin (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), and BA [6-benzylaminiopurine(BA), (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$)] in MS (3% sugar and 0.8% agar with pH 5.7) media. The highest number of induced shoots, leaves and roots were shown in 1/2 MS medium concentration. On the 1/2 MS medium, shoot numbers, shoot length, leaf numbers and root numbers were 11.0, 1.9 cm, 24.7, and 8.0, respectively. On the absence of CPPU in the 1/2 MS medium, shoot length and root numbers was greater than CPPU treatment, but the highest number of shoots was induced by the $2.0mg{\cdot}L^{-1}$ of CPPU concentration in 1/2MS medium. TDZ, zeatin, and BA treatments were not effective on the induction of multiple shoot in vitro culture. As a result, in vitro culture of new Saxifraga fortunei, 'Greenstar' with $2.0mg{\cdot}L^{-1}$ of CPPU in 1/2 MS medium was most effective for the rapid multiplication.

Development of rice(Oryza sativa L.) transformation system to improve callus utilization (캘러스 활용도를 향상시키기 위한 벼(Oryza sativa L.) 형질전환 시스템 구축)

  • Park, Ji-Sun;Moon, Ki-Beom;Ha, Jang-Ho;Jang, Ji-Young;Kim, Mi-Jin;Jeon, Jae-Heung;Park, Sang-Un;Kim, Hyun-Soon
    • Korean Journal of Breeding Science
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    • v.49 no.3
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    • pp.170-179
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    • 2017
  • Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.

Mass Propagation of Hypolepis punctata (Thunb.) Mett. Using In Vitro Culture Techniques (조직배양을 이용한 점고사리의 대량증식 방법)

  • Park, Kyungtae;Jang, Bo Kook;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.57-57
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    • 2019
  • 본 연구는 관상 및 조경용으로 개발이 가능한 남방계 양치식물인 점고사리[Hypolepis punctata (Thunb.) Mett.]의 전엽체 증식 및 포자체 형성에 적합한 배양조건을 구명하고자 수행되었다. 무가온 온실에서 성숙한 포자엽을 채집한 후 포자를 기내에서 발아시켜 전엽체를 획득하였으며, 8주 간격으로 계대배양 하여 실험의 재료로 사용하였다. 배지종류에 따른 전엽체의 증식 및 형태형성을 확인하고자, 배양된 전엽체 0.3g을 메스로 잘게 다진 후, 농도를 1/4, 1/2, 1, 2배로 조절한 MS배지에 8주간 배양하였다. 이후 선발된 배지를 기준으로 sucrose, 활성탄, 질소급원의 농도를 조절하여 전엽체의 증식과 형태형성을 확인하였다. 그 결과, 1MS배지에서 전엽체의 생체중이 초기 접종량인 0.3g에 비해 10.7배 증가한 3.2g으로 가장 높은 증가율을 보였다. 형태관찰에서도 장정기의 형성이 관찰되었으며, 전엽체의 쿠션조직이 비교적 잘 발달하였다. 전엽체 증식에 가장 좋은 효과를 보인 1MS배지를 기준으로 sucrose의 농도를 0-4%로 달리하여 실험한 결과, 1%의 처리구에서 6.7g으로 가장 높은 생체중을 보였다. 활성탄의 농도를 0-0.8%로 첨가한 네 처리구 중에서는 0.8%의 처리구에서 14.2g으로 무처리구에 비해 생체중이 2배 이상 증가하였다. 전엽체의 형태 또한 정상적인 발달을 보였다. 질소급원의 비율을 30-120mM로 조절한 배지에서는 60mM의 처리구에서 4.9g으로 가장 높은 생체중을보였다. 이후 포자체 형성을 위한 최적의 토양조건을 구명하고자, 원예상토, 피트모스, 펄라이트 및 마사토의 비율을 달리하여 5종류의 배양토를 조성하였다. 혼합된 토양은 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진 하였으며, 배양된 전엽체 1g을 증류수와 함께 10초간 분쇄한 다음 준비된 토양표면에 분주하여 재배하였다. 12주간의 재배 결과, 원예상토를 단용한 토양에서 포자체의 수가 포트당 250.0개로 가장 많이 형성되었으며, 포자체의 생육 또한 다른 처리구에 비해 우수한 결과를 보였다. 한편 피트모스가 혼합된 토양에서는 포자체가 형성되지 않았다. 따라서 점고사리의 전엽체 대량증식에 적합한 배지는 sucrose 1%와 질소급원의 농도를 60mM, 활성탄을 0.8% 첨가한 1MS배지로 판단되며, 포자체 대량생산을 위해서는 원예상토를 단용한 토양이 적합하다고 판단된다.

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First Report of Pseudomonas viridiflava Causing Leaf Spot of Cucumber in Korea (Pseudomonas viridiflava에 의한 오이 점무늬병의 발생 보고)

  • Seo, Yunhee;Park, Mi-Jeong;Back, Chang-Gi;Park, Jong-Han
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.328-331
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    • 2018
  • A severe disease with leaf spots and necrotic symptoms was observed in cucumber (Cucumis sativus L.) seedlings in April 2018 at a nursery in Kimjae, Korea (35o 47'09.8"N 127o 2'24.3"E). The infected plants initially showed spots on water-soaked cotyledons which, at later stages, enlarged and spread to the leaves, which the lesions becoming dry and chlorotic. The symptomatic samples were collected from cucumber and the isolates were cultured on LB agar. The representative bacterial strain selected for identification showed fluorescent on King's medium B, was potato rot-positive, levan and arginine dihydrolase-negative, oxidase-negative and tobacco hypersensitivity-positive in LOPAT group 2 as determined by LOPAT tests. A pathogenicity test was carried out on a 3-week-old cucumber. After 3 days of inoculation, leaf spots and necrotic symptoms appeared on the cucumber, similar to the originally infected plants. The infecting bacterial strain was identified as Pseudomonas viridiflava, by 16S rDNA sequence analysis. This is the first report of leaf spot diseases on cucumber caused by P. viridiflava.

Culture condition for gametophyte and sporophyte masspropagation of bamboo fern (Coniogramme japonica) using tissue culture (조직배양을 이용한 가지고비고사리의 전엽체와 포자체 대량번식을 위한 배양조건)

  • Park, Kyungtae;Jang, Bo Kook;Lee, Cheol Hee
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.119-126
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    • 2019
  • This study was conducted to investigate the optimal culture method for gametophyte and sporophyte propagation in Coniogramme japonica (Thunb.) Diels, which can be used in various fields. The propagation of prothallus were cultured in 1/4 - 1 Murashige and Skoog medium and Knop medium for 10 weeks. The results indicated that the fresh weight of prothallus was the highest (14.5 g) in 1MS medium. Subsequently, various concentrations of sucrose, activated charcoal and nitrogen source were also added to 1MS medium and cultured for 8 weeks. The results provided that the sucrose concentration was 3% and the fresh weight of prothallus was the highest 10.8 g. According to the concentration in the range of 8.8 ~ 10.8 g, in the case of activated charcoal, the four treatments showed no significant difference. The nitrogen source was added at a concentration of 30, 60 and 120 mM with the ratio of ${NH_4}^+:{NO_3}^-$ being 1 : 2. As a result, the fresh weight of all treatments increased to similar level but there was no significant difference. We investigated sporophyte formation according to soil type and the highest number of sporophytes at 228.0 was formed in soils mixed with horticultural substrate and decomposed granite at 2 : 1 (v : v). On the other hand, sporophyte was not formed in soils containing peatmoss except for the one with peatmoss and decomposed granite at 2 : 1 (v : v).

Change in the Levels of Intracellular Antioxidants during Aging of Articular Chondrocytes and Cartilage (연골세포 및 관절연골의 노화 과정에서 세포내 항산화 인자들의 변화)

  • Kim, Kang Mi;Kim, Yoon Jae;Kim, Jong Min;Sohn, Dong Hyun;Park, Young Chul
    • Journal of Life Science
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    • v.29 no.8
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    • pp.888-894
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    • 2019
  • Cartilage diseases, such as rheumatoid arthritis (RA) and osteoarthritis (OA), are associated with the loss of chondrocytes and degradation of articular cartilage. Recent studies revealed that inflammatory reactive oxygen species (ROS) and age-related oxidative stress can affect chondrocyte activity and cartilage homeostasis. We investigated changes in the levels of intracellular antioxidants during cellular senescence of primary chondrocytes from rat articular cartilages. Cellular senescence was induced by serial subculture (passages 0, 2, 4, and 8) of chondrocytes and measured using specific senescence-associated ${\beta}$-galactosidase ($SA-{\beta}-gal$) staining. ROS production increased significantly in the senescent chondrocytes. In addition, total glutathione (GSSG/GSH) and superoxide dismutase (SOD) levels and heme oxygenase-1 (HO-1) expression increased in senescent chondrocytes induced by serial subculture. Analysis of changes in intracellular antioxidant levels in articular cartilage from rats of different ages (5, 25, 40, and 72 wk) revealed that total glutathione levels were highest after 40 wk and slightly decreased after 72 wk as compared with those after 25 wk. SOD and HO-1 expression levels increased in accordance with age. Based on these results, we conclude that intracellular antioxidants may be associated with cartilage protection against excessive oxidative stress in the process of chondrocyte senescence and age-related cartilage degeneration in an animal model.

Optimal Culture Conditions for Mass Propagation of Onoclea sensibilis var. interrupta Maxim. (야산고비의 대량증식을 위한 적정 배양조건)

  • Park, Kyungtae;Jang, Bo Kook;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.56-56
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    • 2019
  • 본 연구는 식용과 약용으로 이용되는 국내 자생식물인 야산고비(Onoclea sensibilis var. interrupta Maxim.)를 기내에서 대량증식하기 위한 조건을 구명하기 위하여 수행되었다. 무가온 온실에서 수집한 포자를 기내에서 발아시켜 전엽체를 획득하였으며, 계대배양하며 실험의 재료로 사용하였다. 전엽체의 대량증식을 위해 전엽체 0.3g을 메스로 균일하게 다진 후 증류수 1ml와 함께 배지에 고루 펼쳐서 배양하는 방법을 사용하였으며, 증식에 미치는 배지의 영향을 확인하기 위하여 1/4, 1/2, 1, 2배로 조절한 MS배지를 조성하여 8주간 배양하였다. 이후, 증식이 우수한 배지를 선정하여 sucrose와 질소급원의 농도를 조절하였으며, 활성탄을 첨가하여 증식에 미치는 영향을 확인하였다. 배지종류 실험의 결과, 생체중이 1MS에서 10.2g으로 초기 접종량에 비해 가장 많이 증가하였으며, 현미경을 이용하여 관찰한 결과에서도 정상적인 전엽체의 형태인 심장형으로 발달하였다. 증식이 우수하였던 1MS배지에 sucrose의 농도를 조절하여 배양한 결과에서는 1%의 처리구에서 가장 증식률이 좋았으며, 질소급원의 경우 30mM의 농도로 조절한 처리구가 가장 좋은 결과를 보였다. 배지 내 활성탄의 첨가는 처리구당 증가된 전엽체의 생체중이 유의적인 차이를 보이지 않았다. 포자체 대량 형성을 위한 적정 배양토의 혼합조건을 확인하기 위하여 원예상토, 피트모스, 펄라이트 및 마사토의 혼합비율을 5종류로 달리하여 조성하여 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하였다. 조성한 배양토에 기내에서 배양한 전엽체 1g을 증류수와 함께 블렌더를 이용하여 10초간 분쇄하여 토양표면에 고루 분주하였다. 이후 12주간 재배한 결과, 모든 토양조합에서 포자체가 형성되었다. 그중 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 포트 당 405.0개로 가장 많은 포자체가 형성되었으며, 전체적인 생육 또한 비교적 양호한 결과를 보였다. 따라서 야산 고비의 전엽체 대량증식에 적합한 배지는 경제성과 생육수준을 고려하여 1%의 sucrose와 질소급원의 농도를 30mM로 조절한 1MS 배지가 적합하며, 포자체 대량생산을 위해서는 원예상토, 마사토를 2:1(v:v)로 혼합한 토양이 적합하다고 판단되었다.

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Anti-inflammatory effect of non-thermal atmospheric pressure plasma for periodontitis treatment: in vitro pilot study (치주염 치료를 위한 저온상압 플라즈마의 항염효과: 예비 실험)

  • Park, You li;Kim, Hyun-Joo;Lee, Ju-Youn;Jeong, Sung-Hee;Kwon, Eun-Young;Joo, Ji-Young
    • Journal of Dental Rehabilitation and Applied Science
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    • v.37 no.2
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    • pp.88-94
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    • 2021
  • Purpose: The purpose of this study was to evaluate the anti-inflammatory effects of non-thermal atmospheric pressure plasma (NTP) on human gingival fibroblasts (HGFs) for clinical application of periodontal treatment. Materials and Methods: HGFs were treated with Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS). Customized NTP device was developed for periodontal in vitro study. Cell viability was evaluated with cell counting kit-8. The levels of inflammatory cytokines, including interleukin (IL)-8 and 6, were determined by enzyme-linked immunosorbent assay. Results: When NTP was applied, the cell viability did not change significantly, and there was no difference for 6 h and 24h. When Pg LPS was treated to HGFs, the secretion of IL-8 and IL-6 was increased compared to the control group. But when the NTP was applied, the secretion of them was significantly decreased. Conclusion: NTP did not affect cell viability of HGFs. And it inhibited the LPS-induced production of IL-8 and IL-6.