• 제목/요약/키워드: Long-term subculture

검색결과 16건 처리시간 0.02초

흰제비꽃 배양세포에 있어서 분화세포와 미분화세포 조직의 비교 관찰 (Histological Observation of Embryogenic and Non-embryogenic Callus in Long-term Subculture of Wild Viola (Viola patrinii DC.))

  • 정용모;손병구;이재헌;서정해;정정한;권오창
    • 식물조직배양학회지
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    • 제27권3호
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    • pp.233-238
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    • 2000
  • 야생 흰제비꽃의 엽병 유래 callus를 장기 계대배양하는 과정 중에 발생하는 순화된 friable callus와 분화능이 높은 compact callus를 비교 관찰한 바, friable callus는 연초록색으로 부서지기 쉬운 부드러운 callus이고, compact callus는 진녹색으로 단단한 callus였다. 동결처리 한 시료를 주사전자 현미경에서 동일하게 200배로 관찰하여 보면, friable callus 는 작은 세포집단으로 이루어진 세포군의 주변부에 고도로 액포화된 세포가 광범위하게 분포되어 있는 반면, compact callus는 거의 균일한 세포들로 구성되어 세포구성이 치밀한 것으로 관찰되었다. 또한 friable callus와 compact callus로 부터의 체세포배형성은 배양세포에서 배가 발생하여 배양기간이 지남에 따라 식물체로 분화하였다. 이와 같은 과정은 배양세포의 세포질이 보다 충만한 부위에서 배유사체 (embryo like body)의 발생이 이루어지는 것으로 관찰되었다.

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과잉치 치수유래 줄기세포의 장기 배양 특성 (Characteristics of Supernumerary Tooth-derived Pulp Cells during Long-term Culture)

  • 맹현수;김종빈;김종수
    • 대한소아치과학회지
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    • 제45권4호
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    • pp.492-498
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    • 2018
  • 이 연구는 과잉치 치수로부터 얻은 세포를 더 이상 증식하지 못한다고 판단될 때까지 계대배양을 시행하면서 초기, 중기, 후기 계대의 특성을 비교 분석하였다. 2명의 건강한 6세 남아로부터 3개의 과잉치를 발거하여 치수조직에서 줄기세포를 얻었다. 이를 SNT1(supernumerary tooth 1), SNT2라고 하고, 다른 아이로부터 얻은 과잉치는 SNT3로 명명하였다. SNT1과 SNT2는 똑 같은 시간으로 계대배양 되었고 SNT3은 조금 빠르게 계대배양 되었다. 전체 계대배양의 평균 기간은 $3.6{\pm}1.1$일이었다. 총 $23.3{\pm}0.6$계대까지 배양되었으며, 83일이 소요되었다. 이를 계대를 기준으로 3등분하여 세 군으로 나누었다. I군에서 II군까지 계대배양에 소요된 시간 증가율은 11.90%였으나, II군에서 III군 사이의 증가율은 28.62%로 2.4배가 증가하였다. 22계대까지의 배양기간에 대한 소요 시간을 회귀분석한 결과 y = 0.1169x + 2.25 ($r^2=0.4778$)과 y = 0.1169x + 2.0 ($r^2=0.6444$)으로 추론되었다. 과잉치 치수유래 줄기세포의 정상 배양은 20계대까지 가능할 것으로 판단되었으며, 후기로 넘어가면 계대배양 시간이 2.4배 증가였다. 후기 계대에서는 세포내 부유물이 증가하였고, 형태도 선형으로 변형되는 양상이 관찰되었다.

장기간 계대배양 된 장미 배발생 캘러스로부터 식물체 재분화 및 비정형체로부터 새로운 배발생캘러스 재생 (New embryogenesis from atypical bodies and plant regeneration from long-term subcultured embryogenic callus in rose)

  • 이수영;도경란;천경성;김원희;권오현;이혜진
    • Journal of Plant Biotechnology
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    • 제41권2호
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    • pp.89-93
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    • 2014
  • 장미 형질전환체를 개발하기 위한 유전자 도입 재료로써 5 ~ 6년 이상 장기간 유지 증식된 장미 배발생캘러스의 이용 가능성이 확인되었다. 2007년 및 2009년에 각각 처음으로 기내 뿌리유래 캘러스로부터 유도된 후 유지 증식배지에서 계대배양 해 온 장미 '스위트 옐로우' 품종 및 KR056002와 KR056006 2계통 유래 배발생캘러스로부터 식물체의 재분화 능력을 확인하였다. 장기간 계대배양 된 배발생캘러스로부터 신초의 모습을 갖춘 식물체로 재분화 되기까지 소요기간 및 재분화율은 '스위트옐로우' 품종은 3 ~ 4개월, KR056002 및 KR056006은 4 ~ 5개월로, 이들 배발생캘러스가 기내뿌리로부터 처음으로 배발생 된 후 최초 신초 재분화 때와 동일한 양상이었다. 또한 체세포배발생캘러스의 유지 증식을 위한 계대배양과정에서 발생되는 비정형체 위에 새로운 배 및 배발생캘러스가 유도되었다. 이 비정형체는 새로운 배발생캘러스를 유도할 수 있는 재료로서 이용될 수 있을 것이다.

Laboratory Culture Media-Dependent Biocontrol Ability of Burkholderia gladioli strain B543

  • Bae, Yeoung-Seuk;Park, Kyung-Seok;Choi, Ok-Hee
    • The Plant Pathology Journal
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    • 제23권3호
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    • pp.161-165
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    • 2007
  • Cultivation of a biocontrol agent on a certain medium often results in reduced biocontrol efficacy and alters physiological state. In our previous study, Burkholderia gladioli strain B543 with long-term subculture on tryptic soy agar resulted in significantly reduced biocontrol ability against cucumber damping-off caused by P. ultimum. Therefore, we investigated the influence of laboratory culturing media on biocontrol activity and physiological state of Burkholderia gladioli strain B543 by using long-term repeated culture on a certain medium. When isolate B543 were successionally cultured on King's B agar (KBA), tryptic soy agar, nutrient agar (NA), or soil extract agar more than 20 times, the isolate cultured on KBA or NA showed a significantly enhanced biocontrol efficacy and higher population density in the rhizosphere of cucumber compared to that of the others. However, the isolates cultured on KBA more than 20 times showed the lowest production of protease, siderophore, or antifungal substance(s), measured by skim milk agar, Chrome-Azurol-S agar, and potato dextrose agar amended with 10% of the culture filtrate, respectively. Our results suggest that adaptation to proper culturing medium can alter biocontrol ability and physiological state, and we must consider laboratory media in optimizing the use of biocontrol agents.

Instability of Anthocyanin Accumulation in Vitis vinifera L. var. Gamay Freaux Suspension Cultures

  • Qu Junge;Zhang Wei;Yu Xingju;Jin Meifang
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권2호
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    • pp.155-161
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    • 2005
  • The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, in Vitis vinifera cell cultures. Therefore, four cell line suspensions of Vitis vinifera L. var. Gamay Freaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of $2.73\;\pm\;0.15,\;1.45\;\pm\;0.04,\;0.7\;\pm\;0.024\;and\;0.27\;\pm\;0.04$CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and $84\%$ for V. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be $9.7\%$, ranging from 4 to $17\%$. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities to L-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.

Enhancing in vitro Growth of Bulbs for Mass Propagation of Lily Germplasm

  • Song, Jae-young;Lee, Young-yi;Yi, Jung-yoon;Lee, Jung-ro;Yoon, Mun-sup
    • 한국자원식물학회지
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    • 제34권1호
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    • pp.17-22
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    • 2021
  • Plants regenerated from in vitro cultures carry chromosomal variations, especially in long-term culture. Reducing the duration of plant tissue culture is one of the ways to reduce genetic and epigenetic changes. In this study, we reduced the duration of long-term culture and repeat subculture using small bulblets derived from bulb scales in two lily cultivars. The adventitious bulblets derived from bulb-scale tissue were cultured on three different media containing Murashige and Skoog (MS) basal medium supplemented with 1 g/L Charcoal, MS medium containing 0.3 mg/L IAA and 0.4 mg/L BA hormone with or without Charcoal, respectively. About seven weeks later, the number of newly propagated multiple shoots in the two media, A and B media, showed little differentiation. Compared to both media, the number of propagated multiple shoots increased 5-fold in MS medium containing 0.3 mg/L IAA and 0.4 mg/L BA hormone without Charcoal (C medium). The number of propagated multiple shoots ranged from 5 to 6 and 4 to 6 with an average of 5 in TropicalPink and GreenStar cultivars, respectively. The flow cytometric measurements indicated no variation in the ploidy level between control and in vitro propagated plants.

Brevibacterium ketoglutamicum을 이용한 L-Ornithine 생산 연구 PART I : L-Ornithine 생산 고역가 안정화 균주 선발 (High Production of L-Crnithine by L-Citrulline Auxotroph of Brevibacterium ketoglutamicum : PART I : Selection of Stable Mutant Strains with Less Revertant Formation)

  • 류욱상;장형욱;이홍원;정준기;장순재;유연우;박영훈
    • 한국미생물·생명공학회지
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    • 제27권4호
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    • pp.292-297
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    • 1999
  • Overproduction of L-ornithine by mutant strains isolated from Brevibacterium detoglutamicum BK1046 was investigated. The strain was a L-ornithine auxotroph and exhibited culture instability during fermentation. Through a sequential screening effort, a highly stable strain with lless revertant formation was finally selected and designated B. ketoglutamicum BK52 (KCTC0141BP). It prouduced L-ornithine at a high concentration (above 9 g/L) independent of subculture or cultivation time, and also had a very low tendency of revertant formation. In a long-term storage, this strain maintained its cell stability and productivity of L-ornithine to a reasonable range.

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스팀펑크(steampunk) 패션의 표현 특성 연구 (A Study of Formative Characteristics of Steampunk Fashion)

  • 최유진
    • 복식
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    • 제63권8호
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    • pp.58-75
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    • 2013
  • Steampunk is a term generally used for one of the S. F. subgenre in literature, arts, fashion styles, and lifestyles. This study clarifies formative characteristics as well as the meanings of steampunk fashion style based on its attributes in movies and animations. Attributes of the steampunk are blending time, blending technology, and introspection against science development. In order to have an objective research, this study researched frequency of the fashion item, color, accessory of the collected fashion image, and after interpreted that consequence. Men's steampunk fashion was composed of men's basic outfit such as shirts, vest, jacket, pants, and coats of the 19th century. Black color took up the largest proportion for color. In accessory, top hat, goggle, and steam-powered weapons, in order, were most commonly used. Women's steampunk fashion was composed of shirts and long one-piece dresses, long skirt simply applied 19th victorian style fashion. Yellow, brown, and black color took up the largest proportion of fashion color. In accessory, top hat, belt trimming, corset, and boots, in order, were most commonly used. It is the characteristics of the steampunk fashion that represents retro-future fashion, as it features steam-powered mechanic esthetics, a kind of DIY subculture and transcending division of nationality, class, and gender.

Human Organic Anion Transporting Polypeptide 1B3 Applied as an MRI-Based Reporter Gene

  • Song-Ee Baek;Asad Ul-Haq;Dae Hee Kim;Hyoung Wook Choi;Myeong-Jin Kim;Hye Jin Choi;Honsoul Kim
    • Korean Journal of Radiology
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    • 제21권6호
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    • pp.726-735
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    • 2020
  • Objective: Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and Methods: CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 µL/g). Results: Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion: The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.

Effects of Long-Term Subcultured Arbuscular Mycorrhizal Fungi on Red Pepper Plant Growth and Soil Glomalin Content

  • Selvakumar, Gopal;Yi, Pyoung Ho;Lee, Seong Eun;Shagol, Charlotte C.;Han, Seung Gab;Sa, Tongmin;Chung, Bong Nam
    • Mycobiology
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    • 제46권2호
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    • pp.122-128
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    • 2018
  • Arbuscular mycorrhizal fungi (AMF) are well-known for their ability to improve plant growth and help plants withstand abiotic stress conditions. Unlike other fungi and bacteria, AMF cannot be stored, as they are obligate biotrophs. Long-term preservation of AMF spores is challenging and may lead to the loss of viability and efficiency. This study aimed to understand the effect of prolonged subculture of AMF species on the growth and glomalin-related soil protein (GRSP) from red pepper (Capsicum annuum L.). AMF spores were mass-produced using different techniques and subcultured in pots with sorghum sudangrass as the host plant for 3 years. Experimental soil samples were collected from natural grassland. Five different AMF inocula were used in triplicate as treatments. After 70 days of growth, red pepper plants were harvested and plant dry weight, plant nutrient content, mycorrhizal colonization, AMF spore count, and soil glomalin content were determined. AMF-treated plants displayed higher dry weight than controls, with only fruit dry weight being significantly different. Similarly, significant differences in phosphorous and potassium contents of the above-ground plant parts were observed between mycorrhizal and control treatments. In addition, soil GRSP content was significantly higher in plants inoculated with Rhizophagus sp. and Gigaspora margarita. The increased plant growth and GRSP content suggest that AMF can be maintained for 3 years without losing their efficiency if subcultured regularly with different symbiotic host plants.