• Title/Summary/Keyword: in vitro chromosome doubling

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In Vitro Mitotic Chromosome Doubling by Chemical Treatments in Lilium longiflorum (철포백합의 기내 체세포 염색체 배수화를 위만 화학처리의 효과)

  • Chung, Mi-Young;Chung, Jae-Dong;J. M. VanTuyl;Lim, Ki-Byung
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.73-78
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    • 2004
  • This study was carried out to develop an in vitro chromosome doubling systems for the breeding of tetraploid lily cultivar. Different concentration (ppm) and treatment time (hour) of colchicine, oryzalin and caffeine were compared for the efficiency of bulb regeneration and tetraploidization. The occurrence of tetraploid plants by colchicine was the highest in the concentration of 1,000 ppm for three hours. In oryzalin treatment, the best combination was at 30 ppm for three hours. However, the effects of oryzalin treatments were similar between 10 ppm and 30 ppm concentrations. The survival rate was dramatically reduced in a high concentration of caffeine although some treatments had a higher tetraploid induction. As a consequence, reliable results for the tetraploid production were obtained in the treatments of 1,000 ppm colchicine, 3,000 ppm oryzalin both for three hours, and 9,000 ppm caffeine for nine hours.

Induction of A Chromosome-doubled Persimmon (Diospyros kaki Thunb.) by in vitro Colchicine Treatment (기내 콜히친 처리에 의한 염색체 배가 감 식물체 유기)

  • Ma, Kyeong-Bok;Cho, Kwang-Sik;Jung, Hae-Won;Seo, Ho-Jin;Kang, Sam-Seok
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.515-521
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    • 2018
  • This was carried out to develop a chromosome-doubled (12x) persimmon that will be used as a crossing parent to select seedless persimmon cultivars with the change of the consumption trend recently. To obtain a chromosome-doubled (12x) persimmon, colchicine was applied at the meristem of seedlings in vitro derived from cross among hexaploid persimmon (Diopyros kaki Thunb.). These were treated with 0.03%, 0.05% and 0.1% colchicine respectively for doubling chromosome, and it was most effective at the concentration of 0.05% colchicine. After colchicine treatment, we conducted tests to elucidate conditions for inducing shoot and root development. As the result, the shoots grew best when cultivated at 1/2MS media plus 10 and $30{\mu}M$ zeatin respectively, and the roots grew best when cultivated at 1/2MS media after dipping for 5 seconds at 10 mM NAA+5% DMSO. We also compared seedlings that have chromosome (6x) do not doubled and crossing parents (6x) and chromosome-doubled seedlings (12x). As the result, these chromosome-doubled seedlings (12x) showed lower stomatal density and larger stomatal size.

Study for Metabolism of Resistant Production in Anticancer drug Resistant Stomach Cancer Cell SNU-1 (항암제 내성 위암 세포주 SNU-1의 내성생성기전에 관한 연구)

  • Kim, Jung-Hye;Kang, Mi-Wha;Kim, Jae-Ryong
    • Journal of Yeungnam Medical Science
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    • v.6 no.2
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    • pp.195-205
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    • 1989
  • Development of drug resistance in tumors during treatment is a major factor limiting the clinical use of anticancer agents. When tumor cells acquire resistance to anticancer drug, they show cross-resistance to other antitumor agents. In the present study, SNU-1 cell was induced adriamycin $10^{-7}M$ drug resistance, SNU-1/ADR, in vitro culture system. We got the doubling time and number for viability test during 96 hours by MTT assay. To investigate the cross resistance of various anticancer drugs in human stomach cancer cell SNU-1 and SNU-1/ADR. We compared $IC_{50}$ (drug concentration of 50% reduction) and the relative resistance(RR). SNU-1/ADR was expressed multidrug resistant with vinblastine(RR ; 31.62), vincristine(RR ; 29.50), dactinomycin(RR ; 21.37), epirubicin(RR ; 17.78), daunorubicin(RR ; 14.12), adriamycin(RR ; 7.76), and etoposide(RR ; 4.46), and other drugs, 5-fluorouracil, cisplatin, cyclophosphamide, methotrexate, and aclarubicin, have not cross resistant with adriamycin. There was double minute chromosome in SNU-1/ADR by karyotyping although this change was not seen in SNU-1.

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Use of Androgenesis in Haploid Breeding

  • Yi, Gihwan;Kim, Kyung-Min;Sohn, Jae-Keun
    • Current Research on Agriculture and Life Sciences
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    • v.31 no.2
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    • pp.75-82
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    • 2013
  • Haploids are plants with a gametophytic number of chromosomes in their sporophytes. Androgenesis occurs from asymmetric division of pollen grains into generative cells and vegetative cells, followed by re-entry of the vegetative cell during S-phase, which causes microspores progress into G2/M transition in culture. One of the most interesting features of haploids is the possibility to produce doubled haploid (DH) individuals. Doubled haploidy is extremely useful to plant breeders because it enables shortened breeding periods and efficiency in selection of useful recessive agronomic traits. Doubled-haploid technology is not only applicable to breeding, but also to transformation programs of desired genes. In addition to practical breeding programs, DH lines provide useful materials of fundamental genetics including exploitation of QTLs and genes conferred with various agronomic traits by establishing DH populations. This paper provides historical overviews on androgenesis and describes several mechanisms associated with pollen embryogenesis, including mode of actions in pollen embryogenesis, mechanisms of chromosome doubling and factors affecting androgenesis. We also discuss recent progress in application of haploids to breeding, genes associated with in vitro response and drawbacks to anther culture for application of doubled haploids in crop breeding.

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Chromosome Doubling of Allium wakegi Araki by In Vitro Cultures (기내배양에 의한 쪽파의 체세포 염색체 배가)

  • Yim, Sun-Hee;Ahn, Chang-Soon;Jeong, Chang-Nam;Han, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.259-264
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    • 2002
  • Induction of embryogenic callus from Allium wakegi Araki explants was promoted on medium containing 2,4-D, and production of abnormal embryos from the embryogenic callus increased as 2,4-D concentration was raised. Shoot tip was found to be the best explant source for embryogenic callus formation followed in the order by bulb scale and leaf section. Medium containing 0.09 M sucrose was effective for embryogenic callus production. The regenerants from embryogenic callus on medium containing 2,4-D and BA at different concentrations was consisted of diploids, tetraploids and a few mixaploids of 2n+4n, and their chromosomal aberration rate ranged from 8.0 to 33.3%. Frequency of chromosomal aberrants was the highest (18.7%) in the regenerants obtained from bulb scale-derived embryogenic callus among others. Plant regeneration rate was high (33.5%) from the shoot tip-derived embryogenic callus and the frequency of chromosomal aberrants was very low (7.0%). The plantlets regenerated on medium containing 0.26 M sucrose resulted in high chromosomal aberrants. The regenerants on medium containing sucrose at 0.09∼0.20 M produced chromosomal aberrants at around 15.2∼16.6%.

Establishment and Characterization of Permanent Cell Lines from Oryzias dancena Embryos

  • Lee, Dongwook;Kim, Min Sung;Nam, Yoon Kwon;Kim, Dong Soo;Gong, Seung Pyo
    • Fisheries and Aquatic Sciences
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    • v.16 no.3
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    • pp.177-185
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    • 2013
  • The development of species-specific fish cell lines has become a valuable tool for biological research. In recent years, marine medaka Oryzias dancena has been recognized as a good experimental model fish but there are no reports of establishment of cell lines from this fish. In this study, two cell lines from O. dancena blastula embryos were established from 41 total trials (4.9%). The two cell lines displayed typical in vitro morphology and have been cultured for >121 passages, which corresponds to 293 days. The doubling times of the cell lines were 29.84 and 28.59 h, respectively, and both possessed the potential to expand in a clonal manner, albeit with significant differences between the two cell lines. The absence of any of the four main medium supplements; i.e., fish serum, fetal bovine serum, basic fibroblast growth factor, and medaka embryo extract, significantly inhibited growth. The proportion of cells possessing normal chromosome number was 45% and 46.7% of the cell lines, respectively. Taken together, two cell lines that proliferate continuously were established from marine medaka and these cell lines may provide a basic tool for characterizing the unique features of this fish species.

Response of colchicine for the efficient chromosome doubling in Codonopsis lanceolata

  • Kwon, Soo-Jeong;Lee, Ui Gun;Moon, Young Ja;Cho, Gab Yeon;Boo, Hee Ock;Lee, Moon Soon;Woo, Sun Hee;Kim, Hag Hyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.273-273
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    • 2017
  • Polyploidy has opened a new horizon for selection to sculpt a variety of new gene functions, traits, and lineages. The aim of this study was to investigate the efficacy of the colchicine concentration, temporal changes, and suitable material for inducing effective tetraploid plants of Codonopsis lanceolata. A total of 180 individuals from 16 treatment groups were germinated, and exposed to different concentrations of Colchicine. The plant height of the diploid (18.1 cm) was slightly shorter than that of the tetraploid (13.4 cm). The fresh weight of the main root in the diploid (0.5 g) was 4-fold higher than the tetraploid (2.2 g). The colchicine-treated plant regeneration rate in C. lanceolata was decreased at the elevated concentration of colchicine. A total of 126 individual plants were regenerated in the entire treatment group and tetraploid (2n=4x=32) plants were obtained. In particular, 5 individuals of the tetraploid plant were induced in the 0.05% colchicine for 6h, which is a higher rate (29.4%) than other regenerated plants. As in the seed treatment result, the plant height of the diploid was significantly higher (10.4 cm) than tetraploid. The root length of the tetraploid (10.1 cm) was longer than the diploid, and the root was also thicker. Taken together, the results obtained from the present study may be helpful for the efficient recovery of such polyploid plants through the in vitro application of colchicine, and may improve the productivity and breeding of C. lanceolata.

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Selection of a Triploid Poplar by Flow Cytometric Analysis and Growth Characteristics of its in vitro Grown Plants (유세포 분석을 통한 현사시나무 3배체 선발 및 계통별 기내생장 특성)

  • Bae, Eun-Kyung;Lee, Hyoshin;Lee, Jae-Soon;Noh, Eun-Woon
    • Journal of Korean Society of Forest Science
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    • v.101 no.2
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    • pp.291-296
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    • 2012
  • Triploids are a useful tool for biomass production and molecular breeding of trees with a long life span. Triploids of the poplar 'Hyunsasi' (Populus alba ${\times}$ P. glandulosa) have been developed by crossing between female diploids and a male tetraploid. The tetraploid was developed around the 1970s at Korea Forest Research Institute by colchicine-induced chromosome doubling. Seedlings of the $F_1$ generation were analyzed using flow cytometry to verify their ploidy status. The mean relative fluorescence index of 3 F1 poplars, labeled as Line- 1, Line-17, Line-18, were approximately 1.5 times higher than those of diploid poplars, and the results clearly indicated that they were triploids. The phenotype of the F1 poplars included larger leaves and thicker stem than diploids, and abnormal leaf morphology, especially in the triploid 'Line-18'. Three triploid lines developed roots more slowly and had less roots than diploid. However, 3 poplar cytotypes (2x, Line-1, Line-17) rooted within 10 days on MS medium. In contrast, compared with the 3 cytotypes, the Line-18 showed about 80% and 70% in the rooting rate and the number of roots. The triploid poplars could be directly utilized for biomass production and with their sterility, they could serve as basic material for genetic transformation. In addition, flow cytometric analysis proved to be an effective and reliable method for screening forest trees for their ploidy level.