• Title/Summary/Keyword: Plant cell

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High Frequency Bulblet Formation in Scale and Stem Thin Cell Layer Explant Cultures of Lilium Oriental Hybrids (오리엔탈 백합의 인편 및 줄기의 박판 세포층 절편으로부터 고빈도 자구형성)

  • Oh, Seung-Cheol;Chung, Myung-Hee;Kim, Suk-Won;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.251-255
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    • 2003
  • An efficient system for in vitro bulblet formation of Lilium oriental hybrids(cvs). Casa Blanca and Siberia is described. Transverse thin cell layer(tTCL)(1mm thick) explants of 'Casa Blanca' formed bulblets at a frequency of 97.7% when cultured on Murashige and Skoog (MS) medium supplemented with 1mg/L 2,4 dichlorophenoxyacetic acid(2,4-D) (On average 15.6 bulblets were formed per explant). The frequency of bulblet formation was drastically reduced when the explant ghickness was thinner than 1 mm. Explants from the outermost layer of bulb scale produced greater frequency of bulblet formation than middle or innermost layer. Among auxins supplemented to culture medium at 1 mg/L, 2,4-D led to greater frequency of bulblet formation on explants than dicamba, picamba, or phenylacetic acid(PAA). tTCL explants from the middle region of the outermost layer bulb scale yielded greater frequency of bulblet formation than the upper or lower region. tTCL stem explants of 'Siberia' formed bulblets at a frequency of 95.3% when cultured on MS medium with 1 mg/L 2,4-D(On average 9.1 bulblets were formed per explant). The system estabilished in this study will be useful for in vitro rapid propagation and genetic transformation of Lilium Oriental hybrids.

Effects of Serial Passage on the Characteristics and Chondrogenic Differentiation of Canine Umbilical Cord Matrix Derived Mesenchymal Stem Cells

  • Lee, K.S.;Cha, S.H.;Kang, H.W.;Song, J.Y.;Lee, K.W.;Ko, K.B.;Lee, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.4
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    • pp.588-595
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    • 2013
  • Mesenchymal stem cells (MSCs) are often known to have a therapeutic potential in the cell-mediated repair for fatal or incurable diseases. In this study, canine umbilical cord MSCs (cUC-MSCs) were isolated from umbilical cord matrix (n = 3) and subjected to proliferative culture for 5 consecutive passages. The cells at each passage were characterized for multipotent MSC properties such as proliferation kinetics, expression patterns of MSC surface markers and self-renewal associated markers, and chondrogenic differentiation. In results, the proliferation of the cells as determined by the cumulative population doubling level was observed at its peak on passage 3 and stopped after passage 5, whereas cell doubling time dramatically increased after passage 4. Expression of MSC surface markers (CD44, CD54, CD61, CD80, CD90 and Flk-1), molecule (HMGA2) and pluripotent markers (sox2, nanog) associated with self-renewal was negatively correlated with the number of passages. However, MSC surface marker (CD105) and pluripotent marker (Oct3/4) decreased with increasing the number of subpassage. cUC-MSCs at passage 1 to 5 underwent chondrogenesis under specific culture conditions, but percentage of chondrogenic differentiation decreased with increasing the number of subpassage. Collectively, the present study suggested that sequential subpassage could affect multipotent properties of cUC-MSCs and needs to be addressed before clinical applications.

Rpi-blb2-Mediated Hypersensitive Cell Death Caused by Phytophthora infestans AVRblb2 Requires SGT1, but not EDS1, NDR1, Salicylic Acid-, Jasmonic Acid-, or Ethylene-Mediated Signaling

  • Oh, Sang-Keun;Kwon, Suk-Yoon;Choi, Doil
    • The Plant Pathology Journal
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    • v.30 no.3
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    • pp.254-260
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    • 2014
  • Potato Rpi-blb2 encodes a protein with a coiled-coil-nucleotide binding site and leucine-rich repeat (CC-NBSLRR) motif that recognizes the Phytophthora infestans AVRblb2 effector and triggers hypersensitive cell death (HCD). To better understand the components required for Rpi-blb2-mediated HCD in plants, we used virus-induced gene silencing to repress candidate genes in Rpi-blb2-transgenic Nicotiana benthamiana plants and assayed the plants for AVRblb2 effector. Rpi-blb2 triggers HCD through NbSGT1-mediated pathways, but not NbEDS1- or NbNDR1-mediated pathways. In addition, the role of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) in Rpi-blb2-mediated HCD were analyzed by monitoring of the responses of NbICS1-, NbCOI1-, or NbEIN2-silenced or Rpi-blb2::NahG-transgenic plants. Rpi-blb2-mediated HCD in response to AVRblb2 was not associated with SA accumulation. Thus, SA affects Rpi-blb2-mediated resistance against P. infestans, but not Rpi-blb2-mediated HCD in response to AVRblb2. Additionally, JA and ET signaling were not required for Rpi-blb2-mediated HCD in N. benthamiana. Taken together, these findings suggest that NbSGT1 is a unique positive regulator of Rpi-blb2-mediated HCD in response to AVRblb2, but EDS1, NDR1, SA, JA, and ET are not required.

Somatic Embryogenesis and Plant Regeneration in Mature Seed Cultures and Seed-Derived Embryogenic Suspension Cultures of Yuzu (유자의 성숙종자 배양 및 종자유래 배발생 현탁배양으로부터 체세포배발생을 통한 유자의 식물체 재생)

  • Min, Sung-Ran;Choi, Myung-Suk;Jeong, Won-Joong;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.185-188
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    • 2002
  • Off-white, friable embryogenic calluses were formed on the internal integument of mature seeds of yuzu (Citrus junos) cultured on Murashige and Skoog's basal medium at a frequency of 1.2%. Embryogenic calluses were proliferated when cultured on medium with 1 mg/L 2,4-D. Upon transfer to medium with 0.1 mg/L kinetin, embryogenic calluses produced numerous somatic embryos. Embryogenic suspension cultures were established by placing embryogenic calluses into liquid medium with 1 mg/L 2,4-D. When plated onto medium with 0.5 mg/L ABA, embryogenic cells developed into somatic embryos at a high frequency, and then regenerated into plantlets. Plantlets were successfully transplanted to potting soil and grown in a greenhouse.

Comparison on prevalence and antimicrobial resistance of Pseudomonas spp. isolated from bovine mastitis milk in South Korea (젖소 유방염에서 분리한 Pseudomonas spp.의 분포 및 항생제 내성 비교)

  • Kang, Hye Jeong;Kim, Ha-Young;Hong, Serim;Park, Dasom;Yoon, Soon-Seek;Moon, Jin-San
    • Korean Journal of Veterinary Service
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    • v.44 no.3
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    • pp.133-140
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    • 2021
  • This study was aimed to investigate the prevalence and antimicrobial resistance of Pseudomonas spp. isolated from bovine mastitis milk samples. A total of 50 (4.9%) Pseudomonas spp. was isolated from 1,023 samples, those collected between 2018 and 2021, derived from 110 dairy farms. The prevalence of the identified species of Pseudomonas isolates was as follows; P. aeruginosa (70.0%), P. fluorescens (14.0%), P. putida (10.0%), P. fragi (4.0%), and P. chlororaphis (2.0%). Most of somatic cell counts in the quarter milk carrying Pseudomonas spp. were less than 3,000,000 cell/ml (90.0%). The isolates of Pseudomonas spp. showed high susceptibility to cefepime (98.0%), ciprofloxacin (98.0%), ceftazidime (96.0%), and colistin (96.0%). The rate of antibiotic resistance in the isolates was highest to ceftiofur (92.0%), followed by the resistance rate to chloramphenicol (86.0%) and trimethoprim/sulphamethoxazole (80.0%). In addition, there is a remarkable difference in antimicrobial resistance pattern among Pseudomonas species. P. aeruginosa and P. putida showed a similar resistance pattern, whereas P. fluorescens showed exceptionally lower resistance to trimethoprim/sulphamethoxazole and chloramphenicol than that of the other species. This study showed that prevalence of Pseudomonas spp. other than P. aeruginosa were 30.0% in bovine mastitis milk, and the occurrence rate of antibiotic resistance were similar or higher level, compared with the previous reports on the mastitisderived Pseudomonas spp. isolated in Korea.

Tumorsphere formation and cancer stem cell characterization of REM134 canine mammary carcinoma cells (개 REM134 유선종양세포의 sphere 형성을 통한 암 줄기세포 특성 분석)

  • Byeon, Jeong Su;Lee, Jienny;Jeong, Da-Un;Gu, Na-Yeon;Cho, In-Soo;Cha, Sang-Ho
    • Korean Journal of Veterinary Research
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    • v.58 no.4
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    • pp.201-209
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    • 2018
  • Canine mammary tumors are among the most frequently observed cutaneous tumors in female dogs. Cancer stem cells (CSCs), referred to as tumor-initiating cells, are thought to have properties similar to normal stem cells such as the ability to self-renewal and to differentiate into various cell types. Biological understanding of CSCs and the critical pathways involved in their maintenance are important in research and therapy for mammary tumors. We conducted the present study on sphere formation from REM134 cells by using methylcellulose to produce tumorspheres on a large scale and compared the specific markers of the spheres-formed and plating-cultured REM134 cells. The results revealed that the tumorspheres cultured in methylcellulose had higher seeding density and improved morphology compared to those produced in normal sphere formation medium. Expression levels of stemness markers and CSC-related markers were higher in tumorsphere-forming cells than in plating-cultured cells. Subsequently, we transplanted the tumorsphere-forming and plating-cultured cells into female nude mice to examine their tumorigenic potential. Tumor volume increased rapidly in mice transplanted with tumorsphere-derived cells compared to plating-cultured cells. We observed a novel sphere-forming condition for REM134 cells and showed that REM134 cell tumorspheres can exhibit improved CSC properties.

Disease-resistant Transgenic Arabidopsis Carrying the expI Gene from Pectobacterium carotovorum subsp. carotovorum SL940

  • Lee, Joo-Hee;Hong, Ja-Bin;Hong, Sang-Bin;Choi, Min-Seon;Jeong, Ki-Yong;Park, Hyoung-Joon;Hwang, Duk-Ju;Lee, Seung-Don;Ra, Dong-Soo;Heu, Sung-Gi
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.183-190
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    • 2008
  • Plant-cell-wall-degrading enzymes (PCWDEs) of Pectobacterium carotovorum subsp. carotovorum are the key virulence factor in pathogenesis of soft rot disease of vegetables. The production of PCWDEs is controlled in a cell density dependent manner to avoid the premature production of PCWDEs and subsequent activation of plant defense. N-oxoacyl-homoserine lactone (OHL) is essential for quorum sensing in the soft rot pathogen and the expI gene is responsible for OHL production. The ExpI homolog isolated from P. carotovorum subsp. carotovorum SL940 had 94% identity with ExpI of E. carotovora subsp. carotovora scc3193 and 74% identity with Carl of E. carotovora subsp. atroseptica. The transgenic plants that express exp I uner the control of CaMV35S promoter were able to produce diffusible OHL. Transgenic plants producing OHL were very resistant to the infection of P. carotovorum subsp. carotovorum. Since the PR1 gene was strongly induced and NPR1 and NPR4 were induced weakly in transgenic plants compared to the wild type, salicylic acid-dependent pathways is likely involved in the resistance to the soft rot pathogen P. carotovorum subsp. carotovorum in ExpI transgenic plants.

Distinct Spatio-temporal Expression Patterns of Patatin Promoter-GUS Gene Fusion in Transgenic Potato Microtubers (형질전환 감자 소괴경의 발달단계에 따른 Patatin Promoter-GUS 유전자의 발현 분석)

  • Youm, Jung-Won;Kim, Mi-Sun;Lee, Byoung-Chan;Kang, Won-Jin;Jeon, Jae-Heung;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.13-18
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    • 2003
  • This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

The Effects of Korean Cucurbitaceous Plants on the Alkaline Phosphatase Activity Associated with Sonic Hedgehog Pathway

  • Lee, Hwa Jin
    • Korean Journal of Plant Resources
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    • v.26 no.6
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    • pp.673-677
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    • 2013
  • In order to examine the effects of Korean cucurbitaceous plants on sonic hedgehog pathway and growth of cancer cells with over-activated hedgehog pathway, we measured the sonic hedgehog conditioned medium (shh-CM) induced alkaline phosphatase (ALP) activity and cell viability of pancreatic cancer cell lines by treatment of cucurbitaceous plants. Among the tested cucurbitaceous plants, Actinostemma lobatum Maxim, Cucumis sativus L., Momordica charantia L., Schizopepon bryoniaefolius Maxim and Trichosanthes kirilowii Max, var. japonica Kitam showed the potent inhibitory effects (> 50 % at $20{\mu}g/mL$) on shh-CM induced ALP activity. We also evaluated the cell viability of pancreatic cancer cells treated with the cucurbitaceous plants. The tested cucurbitaceous plants showed the very weak effects on cancer cell proliferation but, T. kirilowii Max, var. japonica Kitam presented the inhibitory effect of 72.7 % on the proliferation of pancreatic cancer cells at $20{\mu}g/mL$. Taken together, we screened the effects of Korean cucurbitaceous plants on shh-CM induced ALP activity and cell viability of pancreatic cancers to search for the modulators of the hedgehog pathway leading to the inhibition of cancer cell proliferation. T. kirilowii Max, var. japonica Kitam, among the tested cucurbitaceous plants, showed the inhibitory effects on the shh-CM induced ALP activity and the proliferation of pancreatic cancer cells.