• Title/Summary/Keyword: polyploid induction

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A Cytogenetic Study on Induction of Diploid Spermatozoa in Poultry (가금류 정자 세포의 배수성 유기를 위한 세포 유전학적 연구)

  • 김철욱;손시환;전익수
    • Korean Journal of Poultry Science
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    • v.23 no.1
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    • pp.1-7
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    • 1996
  • In order to produce polyploid quail, the patterns of spermatogenesis and induction of diploid spermatozoa were analyzed by administration of spindle fiber inhibitor agent. Colcemid at the dose level of 37 $\mu\textrm{g}$ /100 g BW was Injected intraperitoneally to 50 Japanese quail males for 3 consecutive days. Five to 20 days after the first colcemid injection, the metaphase spreads from mitotic spermatogonia, primary spermatocyte and secondary spermatocyte were observed. By cytogenetic analysis, 9.4% of spermatogonia and spermatocyte cells in germ cells from the treated males was found to be polyploid cells. As compared with colcemld treated, the males with non-treated colcemid had only 2.3% polyploid cells in germ cells. The induction of diploid germ cells was highest in 10 days after the first colcemid injection and was lowest in 5 days after the first colcemid injection. These results suggested that between 10 to 15 days before maturation of the spermatozoa, the male germ cells were most sensitive to colcemid treatment. Spindle fiber inhibitor agent was also more sensitive to mitotic division of spermatogonia than meiotic division of primary and secondary spermatocyte.

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Effect of Colchicine on the Induction of Prunella vulgaris for. albiflora Nakai

  • Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Kab-Yeon;Moon, Young-Ja;Woo, Sun-Hee;Kim, Hag-Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.107-113
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    • 2015
  • This study was conducted to find out the effective induction method of tetraploid plants to obtain potential data for cultivating superior varieties by colchicine treatment. The seed germination were decreased by the higher concentration of colchicine treatment and longer soaking time. A total of 907 individuals were germinated in 16 treated plots except control (untreated plot) and 28 tetraploids were induced which was about 3.1% of the number of seed germinated. The plant regeneration rate by colchicine treatment on explant of Prunella vulgaris for. albiflora Nakai under in vitro culture was decreased with the higher concentration of colchicine. While a total of 312 individuals were regenerated in all treatments, the explant was soaked in more than 0.05% for over 1 hour, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 6 hours and 12 hours, 37 tetraploids were induced, which was about 57.8% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploid, the peak DNA content of G1 phase was 101.3 for diploid and 197.2 for tetraploid. The result confirmed the doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 19.3 in tetraploid, which was around 1.9 times as much as diploid.

Effect of Antimitotic Agent Colchicine on In Vitro Regeneration of Watermelon

  • Jaskani Muhammad J.;Raza H.;Khan M. M.;Kwon Sung W.
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.247-252
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    • 2004
  • In vitro cultures of watermelon were treated with antimitotic agent colchicine to induce ploidy alterations, particularly the induction of tetraploids. Explants cotyledon, embryonic end of seed, transverse sections of epicotyl and hypocotyl were cultured on MS media supplemented with BA ($1{\mu}M$) and colchicine ($0.01\%,\;0.05\%\;and\;0.1\%$). Explants were subcultured on colchicine free media after 4 and 7 days. Colchicine had negative effect on in vitro regeneration but this exhibited explants related response. However, hypocotyl section of seedlings induced maximum callus on $0.01\%$ colchicine. Shoot proliferation was more in cotyledon explants cultured on colchicine ($0.01\%$) for four days. Maximum root induction and root number were recorded in embryonic end explants. Overall, cotyledon and embryonic end explants, and low colchicine concentration ($0.01\%$) was found optimal in watermelon regeneration.

Induction of Mutation of Streptomyces and Yeast by Simultaneous Treatment of Electric Shock and Chemical Mutagen N-Methyl-N'-Nitrosoguanidine (전기충격과 화학적 돌연변이원 N-Methyl-N'-Nitro-N-Nitrosoguanidine의 병행처리에 의한 방선균과 효모의 돌연변이 유발)

  • Sun, Jong-Ho;Kim, Jung-Hee;Park, Uhn-Mee;Kim, Keun
    • Korean Journal of Microbiology
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    • v.33 no.3
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    • pp.199-202
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    • 1997
  • The effect of electric shock (AC 38 Vll.3 cm) on the lethal effect and induction rate of mutation of chemical mutagen N-methyl-N'-nitro-N-nitrosoguanidine (NTG) was examined by using the spores of Streptomyces and the cells of haploid and polyploid yeast strains of Saccharomyccrs. Spores of Sireptomyces were all alive after 180 min of electric shock, hut all dead after 960 min-treatment. When the spores of Streptomyces or the cells of haploid and polyploid yeast were treated with electric shock and NTG, the electric shock increased the lethal effect of NTG; the survival rate of Streptomyces dropped from 72 to 48% after 180 min-treatment and those of haploidand polyploid-yeast decreased from 8 to 3% and 25 to lo%, respectively, after 40 min-treatment. The electric shock also increased mutation rates of Streptomyces and haploid yeast. from 1.8 to 13.6%' after 120 min-treatment and from 2.4 to 4.8% after 40 min-treatment, respectively.

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Induction of the Diploid Ovum in Chicken (닭에서 2배수성 난자의 생성에 관한 연구)

  • 여정수
    • Korean Journal of Poultry Science
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    • v.15 no.2
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    • pp.67-71
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    • 1988
  • In order to induce the diploid gamete(ovum) under suppression of meiosis in oogenesis for production of polyploid chicken. this experiment checked meiosis time. through regular ovulation and response of inhibitor (Tri-ethylen Melamine) to meiosis. The results obtained was follows; *Meiosis of oogenesis was 2-4 hours before ovulation. *Response of inhibitor to meiosis was effective at 0.3mg triethylen melamine per kg body weight. *Fertility was highly decreased by influence from inhibitor. *66% of fertilized eggs was triploid(3n) through fertilization induced diploid ovum (2n) with normal sperm(n).

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Microtubule-damaging Chemotherapeutic Agent-mediated Mitotic Arrest and Apoptosis Induction in Tumor Cells (미세소관-손상 항암제 처리에 의한 세포주기의 정지 및 에폽토시스 유도)

  • Jun, Do Youn;Kim, Young Ho
    • Journal of Life Science
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    • v.26 no.3
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    • pp.376-386
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    • 2016
  • Apoptosis induction has been proposed as an efficient mechanism by which malignant tumor cells can be removed following chemotherapy. The intrinsic mitochondria-dependent apoptotic pathway is frequently implicated in chemotherapy-induced tumor cell apoptosis. Since DNA-damaging agent (DDA)-induced apoptosis is mainly regulated by the tumor suppressor protein p53, and since more than half of clinical cancers possess inactive p53 mutants, microtubule-damaging agents (MDAs), of which apoptotic effect is mainly exerted via p53-independent routes, can be promising choice for cancer chemotherapy. Recently, we found that the apoptotic signaling pathway induced by MDAs (nocodazole, 17α-estradiol, or 2-methoxyestradiol) commonly proceeded through mitotic spindle defect-mediated prometaphase arrest, prolonged Cdk1 activation, and subsequent phosphorylation of Bcl-2, Mcl-1, and Bim in human acute leukemia Jurkat T cells. These microtubule damage-mediated alterations could render the cellular context susceptible to the onset of mitochondria-dependent apoptosis by triggering Bak activation, Δψm loss, and resultant caspase cascade activation. In contrast, when the MDA-induced Bak activation was inhibited by overexpression of anti-apoptotic Bcl-2 family proteins (Bcl-2 or Bcl-xL), the cells in prometaphase arrest failed to induce apoptosis, and instead underwent mitotic slippage and endoreduplication cycle, leading to formation of populations with 8N and 16N DNA content. These data indicate that cellular apoptogenic mechanism is critical for preventing polyploid formation following MDA treatment. Since the formation of polyploid cells, which are genetically unstable, may cause acquisition of therapy resistance and disease relapse, there is a growing interest in developing new combination chemotherapies to prevent polyploidization in tumors after MDA treatment.

Improvement of Chromosome Doubling Efficiency in Cymbidium Hybrids by Colchicine and Oryzalin Treatment (Colchicine과 oryzalin 처리에 의한 Cymbidium hybrids의 기내 배수화 효율 증진)

  • Hwang, Sook-Hyun;Kim, Mi-Seon;Park, So-Young
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.900-910
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    • 2015
  • This study aimed to investigate the efficiency of colchicine and oryzalin in inducing polyploidy in two Cymbidium hybrids [Showgirl 'Silky' and Mystery Island 'Silk Road' (Silk Road-4)]. Colchicine was used at concentrations ranging from 50 to $500mg{\cdot}L^{-1}$, with treatments lasting 1 to 3 weeks. Oryzalin was used at concentrations ranging from 3 to $20mg{\cdot}L^{-1}$, with treatments lasting 3 to 6 days or 1 to 3 weeks. The survival rate of PLBs was better in colchicine than in oryzalin solutions. The ploidy levels were screened using flow cytometry. In C. Showgirl 'Silky', the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (60%) and the 2-week treatment in $5mg{\cdot}L^{-1}$ oryzalin (46.7%). In C. Mystery Island 'Silk Road' (Silk Road-4), the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (16.7%) and the 3-day treatment in $10mg{\cdot}L^{-1}$ oryzalin (6.7%). Colchicine was more efficient than oryzalin in terms of polyploidy induction. Furthermore, pre-treatment, which entailed poking 10 times with forceps, improved the efficiency of chromosome doubling.

A Tetraploid Induction in Hypericum patulum Thunberg by Colchicine Soaking Treatment (콜히친 침지처리에 의한 '망종화'의 4배체 식물유도)

  • Kwon, Soo Jeong;Cho, Kab Yeon;Kim, Hag Hyun
    • Korean Journal of Plant Resources
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    • v.26 no.2
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    • pp.284-288
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    • 2013
  • This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. The optimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploid plant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, the higher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. While individuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and no tetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulum Thunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05% for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.

Establishment of an Allo-Transplantable Hamster Cholangiocarcinoma Cell Line and Its Application for In Vivo Screening of Anti-cancer Drugs

  • Puthdee, Nattapong;Vaeteewoottacharn, Kulthida;Seubwai, Wunchana;Wonkchalee, Orasa;Keawkong, Worasak;Juasook, Amornrat;Pinloar, Somchai;Pairojkul, Chawalit;Wongkham, Chaisiri;Okada, Seiji;Boonmars, Thidarut;Wongkham, Sopit
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.711-717
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    • 2013
  • Opisthorchis viverrini (O. viverrini) is a well-known causative agent of cholangiocarcinoma (CCA) in humans. CCA is very resistant to chemotherapy and is frequently fatal. To understand the pathogenesis of CCA in humans, a rodent model was developed. However, the development of CCA in rodents is time-consuming and the xenograft-transplantation model of human CCA in immunodeficient mice is costly. Therefore, the establishment of an in vivo screening model for O. viverrini-associated CCA treatment was of interest. We developed a hamster CCA cell line, Ham-1, derived from the CCA tissue of O. viverrini-infected and N-nitrosodimethylamine-treated Syrian golden hamsters. Ham-1 has been maintained in Dulbecco's Modified Essential Medium supplemented with 10% fetal bovine serum for more than 30 subcultures. These cells are mostly diploid (2n=44) with some being polyploid. Tumorigenic properties of Ham-1 were demonstrated by allograft transplantation in hamsters. The transplanted tissues were highly proliferative and exhibited a glandular-like structure retaining a bile duct marker, cytokeratin 19. The usefulness of this for in vivo model was demonstrated by berberine treatment, a traditional medicine that is active against various cancers. Growth inhibitory effects of berberine, mainly by an induction of G1 cell cycle arrest, were observed in vitro and in vivo. In summary, we developed the allo-transplantable hamster CCA cell line, which can be used for chemotherapeutic drug testing in vitro and in vivo.