• Title/Summary/Keyword: Tetraploid

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Suspension Culture-Mediated Tetraploid Formation in Mouse Embryonic Stem Cells

  • Lee, Jae-Hee;Gong, Seung-Pyo;Lim, Jeong-Mook;Lee, Seung-Tae
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.21-26
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    • 2012
  • Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained $in$ $vitro$ for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

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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    • 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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Production of Tetraploid Embryos with Induction of Diploid Gametes in Chicken (닭에서 2배수성 배우자 생성에 의한 4배수성 생명체의 생산)

  • 여정수;정경진;정익정;정선부
    • Korean Journal of Poultry Science
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    • v.17 no.1
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    • pp.1-6
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    • 1990
  • For the production of tetraploid embryos in chicken through induction of diploid gametes. The experiment was found results as follows; 1. Induction of diploid sperm was observed 11% from embryos of normal females and males at 12 days after injection of 0.37mg per 2kg body weight for three days. 2. 3 among 109 embryos from crosses of diploid gametes induced by TEM were found as tetraploid. 3. Genetic structures of tetraploid embryos were indentified as normal 4 pairs chromosomes without certain variation.

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Induction of Tetraploid Cyprinid Loach, Misgurnus anguillicaudatus (4 배체 미꾸리(Misgurnus anguillicaudatus)의 유도)

  • KIM Dong Soo;NOH Choong Hwan;NAM Yoon Kwan
    • Journal of Aquaculture
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    • v.6 no.1
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    • pp.55-62
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    • 1993
  • A tetraploid cyprinid loach, Misgurnus anguillicaudatus was induced by applying heat shock. The efficiency of inductions changed with the time of initiation of treatment and the duration of treatment. The highest incidence of tetraploid was obtained at the treatment of $41^{\circ}C$ for 3 min, 28 min after fertilization. Ploidy level was estimated by kariological examination and flow cytometric analysis. The tetraploid cyprinid loach had 4n = 100 chromosomes, while the diploid had 2n = 50. The erythrocytic volume of the tetraploid was 1.65 times larger than that of the diploid. The DNA content of the tetraploid and diploid showed 6.547 pg/cell and 3.067 pg/cell, respectively.

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Comparison of Yield and Growth Characteristics of Platycodon grandiflorum According to the Ploidy Levels and Growing Conditions

  • Lee, Hee-Doo;Seo, Dong-Yeon;Kwon, Soo-Jeong;Kim, Hyun-Jeong;Cho, Gab-Yeon;Boo, Hee-Ock;Roy, Swapan Kumar;Kim, Hag-Hyun
    • Korean Journal of Plant Resources
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    • v.29 no.3
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    • pp.331-338
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    • 2016
  • The present study aimed to compare and investigate the morphological characteristics and yield components according to ploidy level of diploid and tetraploid Platycodon grandiflorum under vinyl-greenhouse and open field conditions. Plant height of diploid and tetraploid P. grandiflorum was 51.3 ㎝, 54.0 ㎝, respectively. The results revealed that the plants grown in the vinyl-greenhouse showed significantly higher growth compared to those grown in the open field. Regardless of the growing place, diploid and tetraploid of P. grandiflorum showed the rapid elongation of internodes after 4 and 3 internodes respectively and elongation tends to be decreased as entering the flower-bud differentiation period. The starting day of flowering in vinyl-greenhouse cultivation was found to be faster than that of the open field cultivation by 2∼3 days and tended to be delayed by about 5∼6 days in tetraploid P. grandiflorum compared to diploid. Fresh weight of roots from the vinyl-greenhouse cultivation showed a high quantity as 34.2g and 49.4g in diploid and tetraploid P. grandiflorum, respectively and especially tetraploid P. grandiflorum was found to be increased by approximately 44.4% compared to other plots.

Incidence of Severe Crown Gall Disease on Tetraploid Cultivars of Grape in Korea

  • Park, Kwang-Hoon;Jeong, Kyu-Sik;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.290-293
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    • 2000
  • The main grape producing regions in Korea were surveyed for the occurrence and relative incidence of crown gall disease on grapevine. The results of the survey showed that the disease tended to affect tetraploid cultivars, which produce a large and sweet fruit but are very weak to cold weather. Incidence of crown gall disease was high on the tetraploid cultivars, Kyoho (Gerbong), Daebong, and Black Olympia while it was low on the diploid cultivars, Cambell Early and Sheridan. The disease incidence was very high on Anseong, Cheonan, and Chincheon, where the major growing areas of tetraploid cultivars and grapevines were burried in the winter to protect from freeze injury, whereas it was low in Yungdong, Kimcheon, and Nonsan. Crown gall disease did not increased with grapevine age on both Cambell Early and Kyoho. It remained low in Cambell Early, but high for all ages on Kyoho, Galls were found on the crown region and mid part of the trunk, but more galls were on small branches and canes on Kyoho grapevines. More than one third of ZKyoho grapevines inspected had galls on multiple locations on grapevines. On Kyoho, 56.3% of the galls covered more than 50% of the crown gall disease affects severely on the cultivar.

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A Comparison between Diploid and Tetraploid Cultivars of Lolium multiflorum Lam, italicum (이탈리안 라이그라스의 2배체와 4배체 품종간 비교)

  • 박병훈;박병식;강정훈
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.7 no.3
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    • pp.135-139
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    • 1987
  • The influence of ploidy level on agronomic characteristics in italian ryegrass, Lolium multiflorum Lam, italicum, was studied using diploids (2x) and tetraploids (4x) on the upland of Livestock Experiment Station, Suweon. The results are summarized as follow: 1. Tetraploid cultivars are in general dark green in leaf color and strong glittered. 2. Diploid cultivars are more resistant to cold than tetraploids but degree of resistance to cold depends on cultivars. The di-and tetraploids did not differ in summer survival and coldand heat resistance of di-Itetraploid cultivars were not related. 3. Tetraploid cultivars have larger leaf-blades, more rapid growth and higher leaf ratio to stem by heading than diploids. 4. Tetraploids possess significantly lower dry matter content than diploids. 5 . Fresh- and dry weight of tetraploids are similar with that of diploids but tetraploids are more leafy than diploids. 6. Seasonal yield depends more on the type of the cultivar than on its tetraploid nature.

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Studies on a Tetraploid of Robinia pseudoacacia L. Selected in the Planted Forest (조림지(造林地)에서 선발(選拔)한 자연(自然) 4배체(倍體) 아까시나무(Robinia pseudoacacia L.)에 관(關)한 연구(硏究))

  • Kim, Chung Suk;Lee, Suk Koo
    • Journal of Korean Society of Forest Science
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    • v.14 no.1
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    • pp.39-44
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    • 1972
  • Cytological investigation was done on a tetraploid tree of Robinia pseudoacacia L. selected in the planted forest in Chilbo Mountain. From the results of observation of chromosomes on mitotic metaphase in leaves, it was confirmed that the mutant is tetraploid plant (2n=40). The external form of leaves of the spontaneous tetraploid was gigantic and dark green one in comparison with diploid. There was no difference in the size of guard cell and the number of stomata per unit area between the mutant and the diploid plant, however, length and thickness of wood fiber were increased as compared with that of diploid. The spontaneous gigas tetraploid showed growth performance as much as the colchicine induced gigas tetraploid.

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Generation of Embryonic Stem Cell-derived Transgenic Mice by using Tetraploid Complementation

  • Park, Sun-Mi;Song, Sang-Jin;Choi, Ho-Jun;Uhm, Sang-Jun;Cho, Ssang-Goo;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • pp.121-121
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    • 2003
  • The standard protocol for the production of transgenic mouse from ES-injected embryo has to process via chimera producing and several times breeding steps, In contrast, tetraploid-ES cell complementation method allows the immediate generation of targeted murine mutants from genetically modified ES cell clones. The advantage of this advanced technique is a simple and efficient without chimeric intermediates. Recently, this method has been significantly improved through the discovery that ES cells derived from hybrid strains support the development of viable ES mice more efficiently than inbred ES cells do. Therefore, the objective of this study was to generate transgenic mice overexpressing human resistin gene by using tetrapioid-ES cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR and cloned into pCR 2.1 TOPO T-vector and constructed in pCMV-Tag4C vector. Human resistin mammalian expression plasmid was transfected into D3-GL ES cells by lipofectamine 2000, and then after 8~10 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 $\mu$sec. (fusion rate : 93.5%) and cultured upto the blastocyst stage (development rate : 94.6%). The 15~20 previously G418-selected ES cells were injected into tetraploid blastocysts, and then transferred into the uterus of E2.5d pseudopregnant recipient mice. To investigate the gestation progress, two El9.5d fetus were recovered by Casarean section and one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, this finding demonstrates that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mouse for the rapid analysis of gene function in vivo.

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Cold-hardiness Tetraploid Induced by Colchicine Treatment in Mulberry Seedings(Morus alba L. Yongchonppong/Kaeryanppong) (지배실생(용천뽕/개량뽕)의 생장점에 Colchicine 처리로 창성한 내동성계 4배체뽕)

  • 박광준
    • Journal of Sericultural and Entomological Science
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    • v.36 no.1
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    • pp.1-7
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    • 1994
  • Four tetraploid mulberry lines, Sawonppon No.11, No.13 and No.14, were induced by the colchicine treatment on the shoot tips of seedlings originated from the cross between Yongchonppong and Kaeryanppong. The major characteristics of the tetraploid lines were as follows: Green tip sprouting stage was similar to Kaeryanppong, a medium budding variety. Leaves were medium- to large-sized cordate type. Leaf surface was rougher and stronger than that of the diploid parents. Leaf thickness, leaf area weight and leaf water content were higher than those of the diploid parents. Average branch length was shorter than that of the diploid parents. Internode length and number of lateral branches were similar to the average values of the two parents. Death atop rate of branch was 1.6%~2.5% indicating strong cold-hardiness of the tetraploid lines. Therefore, these lines could be used as sources of cold-hardiness in developing triploid lines.

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