• Title/Summary/Keyword: Metaphase chromosome

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A Cytological Study on Two Species of Genus Semisulcospira ( Gastropoda: Pleuroceridae ) in Korea (한국산 다슬기류 ( Semisulcospira spp. ) 두 종의 세포학적 연구)

  • 김성순;김동찬;정평림;정용재
    • The Korean Journal of Malacology
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    • v.3 no.1
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    • pp.24-34
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    • 1987
  • The melaniid snails belonging to genus Semisulcospira were collected in the Kangwha and Yonchon areas of Korea in 1986 through 1987 in order to carry out a cytotaxonomic study, The snails were first narcotized with menthol and fixed with 70% ethyl alcohol for morphological identification. The gonads of adult snails were used for chromosome analyses by the technique of Imai et al. (1977) with minor modification. Slide preparations were observed under high power fields using a Leitz light miscroscope. The results obtained in the present stuedy are summarized as follows: 1)The sanils collected from Kangwha and Yonchon areas were identified as Semisulcospira forticosta(Martens, 1886)and S. gottschei (Martens, 1886) respectively.2)No specific differences were obwerved in details of the chromosome cycle between S. forticosta and S. gottschei.3) Diploid chromosome numbers observed at mitotic metaphase were 36. There was no difference in chromosome numbers between S. forticosta and S. gottschei.4) There were morphological differences in the karyotypes of the two species. The spermatogonial metaphase karyotype of S. forticosta consists of six pairs of metacentric, eleven pairs of submetacentric, and one pair of acrocentric chromosomes. The spermatogonial metaphase karyotype of S. gottschei consists of five pairs of metacentric, tselve pairs of submetacentric, and one pair of acrocentric chromosomes. Summarizing the aboxe results, the two species of Semisulcospira employed in this study have same chromosome numbers(2n=36)with different karyotypes.

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Karyotype Analysis of Tobacco Moth, Ephestia elutezza H. (Lepidoptera : Pyralidae) and Cigarette Beetle, Lasioderma serricorne F. (Coleoptera : Anobiidae) (다색알락명나방(Ephestia elutella H.)과 궐련벌레(Lasioderma serricorne F.)의 핵형 분석)

  • Chae, Soon-Yong;Kim, Sang-Seock;Chang, Young-Dock
    • Journal of the Korean Society of Tobacco Science
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    • v.18 no.1
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    • pp.30-38
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    • 1996
  • Meiotic metaphase chromosomes of the testis of two storage insects, tobacco moth (Ephestia elutella H.) and cigarette beetle (Lasioderma serricorne F.) were examined to study their karyotypes. The number of haploid chromosomes of the tobacco moth was 31 and the karyotype was characterized by 29 metacentric and 2 submetacentric chromosomes. The metaphase chromosome length of tobacco moth ranged ca. 1.44-2.11${\mu}{\textrm}{m}$ and the average length was 1, 78${\mu}{\textrm}{m}$. The number of haploid chromosomes of the cigarette beetle at metaphase was 10 and the karyotype was characterized by 8 metacentric and 2 submetacentric chromosomes. The range of metaphase chromosome length of the cigarette beetle was ca. 1.79- 2.39${\mu}{\textrm}{m}$ and the average length 2.09${\mu}{\textrm}{m}$.

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Plant genome analysis using flow cytometry

  • Lee Jai-Heon;Kim Kee-Young;Chung Dae-Soo;Chung Won Bok;Kwon Oh-Chang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 1999.05a
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    • pp.162-163
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    • 1999
  • The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.

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Development of an Efficient Method for Obtaining Metaphase Chromosomes in Individual Blastomeres of Mouse and Human Preimplantaion Embryos: Effect of Microtubule Depolymerizing Agents (착상전 배아의 분리된 할구에서 중기염색체 상을 획득하기 위한 효율적인 방법의 개발에 대한 연구: 미세소관 형성 저해제의 효과)

  • Lim, Chun-Kyu;Min, Dong-Mi;Lee, Hyoung-Song;Kim, Jin-Young;Koong, Mi-Kyoung;Kang, Inn-Soo;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.2
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    • pp.111-118
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    • 2003
  • Objectives: The development of an useful method for obtaining metaphase chromosomes from a biopsied blastomere would allow differentiation between embryos with balanced and normal chromosome complements in the preimplantation genetic diagnosis for chromosomal translocations. This study was performed to evaluate the effects of microtubule depolymerizing agents (MTDAs) on the blastomeres of mouse and human preimplantation embryos, and to establish an effective method for obtaining metaphase chromosomes of biopsied blastomeres in human early embryos. Materials and Methods: Early embryos (2-4 cell stage) from superovulated mice (ICR strain) were collected and treated with single or mixture MTDAs, such as vinblastine, nocodazole and colcemid. After the treatment of MTDAs for 16 hours, the metaphase aquisition (MA) rates were evaluated by the observation of chromosome status with bis-benzimide or DAPI staining. The optimal condition from the above experiment was applied to human embryos, which were developed from abnormal fertilization (3-pronuclei). Fluorescence in-situ hybridization (FISH) with whole chromosome probes was conducted on the human metaphase chromosomes by the MTDAs. Results: In mouse embryos, the effective concentrations of each MTDAs for obtaining metaphase chromosomes were $1.0{\mu}M$ of vinblastine (20.3%), $5.0{\mu}M$ of nocodazole (28.1%) and $1.0{\mu}M$ colcemid (55.6%), respectively. The highest MA rate (91.2%) in the mouse embryos was obtained by a mixture of vinblastine ($1.0{\mu}M$) and nocodazole ($1.0{\mu}M$). In the human embryos, the metaphase chromosomes of blastomeres were obtained in 44 of 113 blastomeres (38.9%) by treatment of the mixture of vinblastine and nocodazole. FISH signals of the metaphase chromosomes were successfully observed in human individual blastomeres. Conclusions: The treatment of a mixture MTDAs for obtaining metaphase chromosomes was an efficient method, and the MA rate was above 90% in the mouse embryos. However, only a relatively small proportions of the blastomeres yielded metaphase chromosomes by the MTDAs in the human embryos. The inconsistent effects of MTDAs may be related to the variation of different species and the poor developmental potency of abnormally fertilized human embryos. We should develop more reliable and efficient methods for obtaining the metaphase chromosomes in the biopsied blastomeres of human preimplantation embryos.

Analysis of Chromosome Composition of Gastrodia elata Blume by Fluorescent in situ Hybridization using rDNA and Telomeric Repeat Probes (rDNA와 말단소체 반복서열 탐침을 이용한 천마의 FISH 염색체 조성 분석)

  • Zhou, Hui Chao;Park, Eung Jun;Kim, Hyun Hee
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.113-118
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    • 2018
  • Background: Gastrodia elata Blume is a saprophytic perennial plant in the Orchidaceae family, because of its agricultural and medicinal effectiveness, researchers focus on its genome and chemical components. However, cytogenetic information based on the chromosome structure and composition to construct chromosomal backbone for genome sequencing research and for the development and breeding of plants is very limited. Methods and Results: We determined the metaphase chromosome composition of the G. elata genome by fluorescence in situ hybridization (FISH) using 5S and 45S rDNAs and telomeric repeat probes. The nuclear genome of G. elata was organized into 2 n = 36, with relatively small ($2.71-5.50{\mu}m$)chromosomes that showed gradual decrease in size. Conglutination phenomenon was observed among the metaphase chromosomes, and it was distinguished from that in other plant metaphase chromosome spreads. One pair of signal was detected for each 5S and 45S rDNA in the pericentromeric region and interstitial region on the short arm of chromosomes 10 and 4, respectively, and telomeric DNA signals were detected in the terminal region of most chromosomes. Conclusions: To our knowledge, this is the first FISH chromosome composition result in G. elata and could be useful in more comprehensive molecular cytogenetic and genomic analyses as well as breeding programs of the medicinal plant G. elata.

Effect of Mitogens, Supplement of β-mercaptoethanol and Fetal Bovine Serum Supplementation in Whole Blood Culture Medium for Bovine Chromosome Analysis

  • Takagi, M.;Tsuno, M.;Ohtani, M.;Acosta, T.J.;Miyamoto, A.;Miyazawa, K.;Sato, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.5
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    • pp.615-621
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    • 2002
  • We investigated the effects of the mitogen supplements of 3 types, pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (ConA), to a whole blood culture system on the number of metaphase spreads obtained in perinatal bovine chromosome analysis. In addition, the supplementation of ${\beta}$-mercaptoethanol (${\beta}$-ME) and FBS was examined in such system. Significant differences (p<0.05) were seen in the number of metaphase spreads with PHA stimulation compared to both PWM and ConA stimulation. When examined the effects of ${\beta}$-ME supplementation, the number of metaphase spreads was significantly (p<0.05) increased at $30{\mu}M$ ${\beta}$-ME compared to control. When evaluated FBS supplementation during PWM stimulation, no significant effect of the supplementation was found. Finally, the effects of the cortisol concentration (10-20, 20-30 and >30 ng/ml) of the blood samples were examined. There was no significant effect of cortisol concentration (p>0.05) among these 3 cortisol concentration groups. The mean percentages of normal metaphase plates (2n=60) from each calf 1) with ${\beta}$-ME, 2) without ${\beta}$-ME and 3) with FBS stimulated with PWM were not significantly different (p>0.05). In conclusion, these findings may be useful in cytogenetic screening programs for not only perinatal calves but also for mature cattle.

Cytogenetic Analysis Using Mitosis, Meiosis Chromosomes and bicolor Fluorescence in situ Hybridization of Bupleurum latissimum Nakai (체세포분열과 감수분열 및 bicolor FISH를 이용한 섬시호의 세포유전학적 분석)

  • Kim, Soo-Young;Bang, Jae-Wook;Lee, Joong-Ku
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.6
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    • pp.354-359
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    • 2006
  • Chromosome analysis using mitosis, meiosis and bicolor FISH were carried out in Bupleurum latissimum Nakai, which is one of the endemic plants in Ulleung island of korea. The somatic methaphase chromosomes number of this plant was 2n = 2x = 16 and the chromosome complements consisted of six pairs of metacentrics and two pairs of submetacentrics. The size of chromosomes ranged 2.40${\sim}$4.20 ${\mu}$m and NOR (nucleolus organizer region) chromosome did not observed using conventional staining. In meiosis chromosomes, metaphase-I and anaphase-I were observed. Metaphase-I anaphase-I showed 8 bivalents and chromosomes migration to make two daughter cells. Using bicolor FISH, one pair of 5S and 45S rDNA signals were detected on the centromeric region of chromosome 3 and the end of short of chromosome 2,respectively. We also observed the NOR using 45S rDNA probe.

Chromosome Study on the Oxyloma hirasei ( Gastropoda : Succineidae ) in Korea (한국산 뾰족쨈물우렁이 ( Oxyloma hirasei ) 의 염색체 연구)

  • 박갑만;김재진
    • The Korean Journal of Malacology
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    • v.13 no.2
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    • pp.125-130
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    • 1997
  • The mitotic and meiotic chromosomes of Succineidae snail one species, Oxyloma hirasei(Pilsbry), were investigated by mians of air-drying mithod. The diploid chromosome numbers were n=18, 2n=36. Chromosome complements of this species consist of seven pairs of metacentrics and 11 pairs of submetacentric chromosomes. Spermatogonial metaphase chromosomes range in length from 4.80${\mu}{\textrm}{m}$ for the largest pair to 1.44${\mu}{\textrm}{m}$ for the smallest pair.

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Studies on Meiosis of PMC's in P. alba × glandulosa and Their Parents (P. alba × glandulosa와 그 양친(両親)의 Pollen Mother Cell의 Meiosis에 관(關)한 연구(硏究))

  • Cheung, Hyon Pae;Chon, Sang Kun;Kim, Mal Sook;Kim, Chung Suk
    • Journal of Korean Society of Forest Science
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    • v.45 no.1
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    • pp.51-61
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    • 1979
  • The chromosome behavior and it's synapsis in the meiosis of pollen mother cell were studied on Populus alba L. as a female parent tree, Populus glandulosa Uyeki as a male parent tree and their hybrid, Populus alba x glandulosa. 1. At metaphase I, the number of nuclear plates with early separation chromosome were observed with the lowest proportion of 11.0% in Populus glandulosa and with the highest proportion of 13.0% in Populus alba${\times}$glandulosa. 2. At metaphase II, early separation chromosomes appeared with the frequency of 11.0% in Populus alba${\times}$glandulosa. But the frequency was not different with those of the parental trees. 3. At anaphase I, lagging chromosomes appeared with some high rate of 11.6% in Populus alba${\times}$glandulosa and yet the number of chromosome bridges in populus alba x glandulosa almost were not different with the partental trees. 4. At anaphase II, lagging chromosomes appeared with some high frequency of 10.2% in Populus alba${\times}$glandulosa and the chromosome bridges in Populus glandulosa appeared with the highest frequency in all studied trees. 5. The frequency of abnormal pollen sporad was the highest value of 8.2% in Populus alba${\times}$glandulosa among the studied trees. With the results, it might be assured that the chromosome segregation and it's synapsis behaved normally in Populus alba, Populus glandulosa and Populus alba x glandulosa, and so all the studied trees could produced normal pollens.

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Chromosomal Analysis of Meiosis and Mitosis in New Zealane White Rabbit (New Zealand White 토끼의 생식세포 및 체세포 분열에 의한 염색체 분석)

  • 신선희;김희수;최영현;이원호
    • Journal of Life Science
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    • v.11 no.4
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    • pp.354-361
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    • 2001
  • Chromosomal characteristics of New Zealane White rabbit was studied at meiosis and mitosis. The meiotic chromosomal preparations were mad with the modified air-drying method and karyotype analysis was performed with the G-banding technique, using isolated mitotic metapase chromosomes of the New Zealand White rabbit. Chromosomes, sex vesicles and centromeres could be classified in the zygotene and the pachytene of the meiosis I. The hair-like processes projecting laterally from the axes of bivalent chromosomes at the mid-to-late pachytene were observed and made the appearance of the lampbrush chromosome structure. Chromosomes could be classified onthe basis of the numbers and locations of chiasma in the diakinesis. Twenty-one autosomal bivalents and a single unequal terminally associated X-Y bivalent were observe during the late prophase and the metaphase of the meiosis I. Most of the bivalent types observed in the New Zealand White rabbit spermatrocytes were 1CH, 1TAl, and 2TA bivalents. The mean chiasma frequency(CF) of the male New Zealand White rabbit was 30.2 and it was found that the CF value tended to decrease through diakinesis and the metaphase I. The karyotype of the New Zealand White rabbit was a male chromosome number of 44(2n=44) comprising 8 pairs of metacentric, 9 pairs of submetacentric, 4 pairs o acrocentric autosomes, metacentric X chromosome and acrocentric Y chromosome.

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