• 제목/요약/키워드: haploid

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In Vitro Development and Apoptosis in Haploid, Diploid Parthenotes and Fertilized Embryos

  • Y. J Chung;Lee, H. Y.;S. H. Jun;X. S. Cui;Kim, N. H.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.28-28
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    • 2003
  • Haploid parthenotes have been shown to be developmentally delayed compared with diploid parthenogenetic embryos in the mouse and pig. These developmental defects have been hypothesized to rusult from insufficient parthenogenetic activation, suboptimal in vitro culture conditions, or genemic imprinting. In the present study we compared the incidence of apoptosis and apoptosis related gene expression in pig haploid, diploid parthenotes and fertilized embryos. In vitro matured porcine oocytes were activated by electrical stimulation. Haploid activated oocytes with two polar bodies under stereomicroscopy were defined haploid parthenotes, oocytes with one polar body were defined as diploid parthenotes after 3h cycloheximide teatment. The morphological analysis of apoptosis in embryos was carried out using propidium iodide staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. The expression of Bcl-xL, Bak and P53 in haploid, diploid and in vivo fertilized blastocysts was determined using RT-PCR. Lower number of the haploid pig parthenotes developed to the morulae and blastocysts compared to the diploid parthnotes. Number of cells significantly lower in the haploid-derived blastocysts than diploid-derived it. Developmentally retarded haploid parthenotes exibited apoptosis at a significantly higher frequency than did diploid parthenotes and fertilized embryos. Level of Bcl-xL expression, diploid parthenotes similar to in vivo-derived it was higher than haploid parthenotes. However, Bak and P53 mRNA expression were not different among haploid, diploid, and fertilized embryos. This result suggested that parthenogenetic activation and parthenogenesis themselves do not cause apoptosis, but haploid increases the incidence of apoptosis in preimplantation embryos. Apoptosis may be due to decrease expression of Bcl-xL in haploid parthenotes developing in vitro.

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Inhibitors Targeting ABA Biosynthesis and Catabolism Can Be Used to Accurately Discriminate between Haploid and Diploid Maize Kernels during Germination

  • Kwak, Jun Soo;Kim, Sung-Il;Song, Jong Tae;Ryu, Si Wan;Seo, Hak Soo
    • Plant Breeding and Biotechnology
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    • 제5권3호
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    • pp.204-212
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    • 2017
  • There is a growing preference for using doubled haploids (DHs) in maize breeding programs because they reduce the time required to generate and evaluate new lines to 2 years or less. However, there is an urgent need for efficient techniques that accurately discriminate between haploid and diploid maize kernels. Here, we investigate the effects of several hormones and chemicals on the germination of haploid and diploid maize kernels, including auxin, cytokinin, ethylene, abscisic acid (ABA) biosynthesis inhibitor (fluridone), ABA catabolism inhibitor (diniconazole), methyl jasmonate (MeJA), and NaCl. Ethylene effectively stimulated the germination of both haploid and diploid maize kernels. The ABA biosynthesis inhibitor fluridone, the ABA catabolism inhibitor diniconazole, and MeJA selectively stimulated the germination of haploid maize kernels. By contrast, gibberellin, 1-naphthaleneacetic acid (NAA), kinetin, and NaCl inhibited the germination of both haploid and diploid maize kernels. These results indicate that the germination of haploid maize kernels is selectively stimulated by fluridone and diniconazole, and suggest that ABA-mediated germination of haploid maize kernels differs from that of diploid maize kernels and other plant seeds.

반수체 넙치(Paralichthys olivaceus)의 발생학적 특성 분석 (Embryonic Development of Haploid Olive Flounder Paralichthys olivaceus)

  • 정효선;김연경;김현철;노재구;이정호;김동수
    • 한국수산과학회지
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    • 제48권5호
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    • pp.696-703
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    • 2015
  • We investigated the characteristics of embryonic and abnormal organ development in haploid olive flounder, Paralichthys olivaceus, by comparing egg development and histological sections in haploid and diploid individuals. After the mid-blastula transition, abnormal development was observed in haploid fish, including delayed epiboly and malformation of the germ ring and embryonic body. In haploid flounder, Kupffer’s vesicles are irregularly shaped and of variable size compared to diploids. The embryonic body of haploids was shorter and broader than that of diploids and the tail length and size were variable. Most haploid embryos failed to hatch and the few larvae that did, did not survive for more than 6 h. The histological analysis of haploid larvae revealed deformed development in diverse organs, including the eye, otic vesicles, notochord, and neural tube. These results may be related to an abnormality in the axial system of haploid larvae. This study confirmed that the abnormalities of haploid olive flounder were similar to the reported characteristics of haploid syndrome. The abnormalities are caused by delayed epiboly and involution and deformity of Kupffer’s vesicle during egg development.

연초의 이배체와 반수체의 저온단일 감응성 비교 (Comparison of Diploid and Haploid Plants for Cool Temperature and Short - day in Nicotiana tabacum L.)

  • 정윤화;금완수
    • 한국연초학회지
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    • 제16권2호
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    • pp.108-112
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    • 1994
  • Response of two ploidy levels to cool temperature and short day treatment were investigated under controlled conditione of Phytotron. The haploid and diploid of seven genotypes were started and grown to the 8- leaf stage in the greenhouse. They were treated during 15 and 20 days to 8- hour photoperiods at 18$^{\circ}C$ in controlled - environmental room to induce premature flowering, respectively. Diploid plants of seven genotypes flower later than their haploid plants at 20 days treatment. But under 15 days treatment, diploid plants of NC82, Hicks, BY4, NC2326 and Coker86 were not different from their haploid plants for days to flower. Diploid plants of seven genotypes developed the same number of leaves as their haploid plants at 20 days treatment. Under 15 days treatment, diploid plants of Coker347 and NC95 developed more leaves per plant than their haploid plants. Correlation coefficient between the ranks of leaves per plant of seven genotypes at two ploidy levels was 0.964 and 0.929 at 15 and 20 days treatment, respectively.

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Efficient method for generating homozygous embryonic stem cells in mice

  • Kim, Bitnara;So, Seongjun;Choi, Jiwan;Kang, Eunju;Lee, Yeonmi
    • 한국동물생명공학회지
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    • 제37권1호
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    • pp.48-54
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    • 2022
  • Parthenogenesis is maternally uniparental reproduction through the embryonic development of oocytes without fertilization. Artificial activation of mature oocytes could generate homozygous haploid embryos with the extrusion of the second polar body. However, the haploid embryos showed low embryo development in preimplantation embryos. In this study, we investigated whether the electronic fusion of the haploid embryos could enhance embryo development and ESC establishment in mice. Haploid embryos showed the developmental delay from 4-cell to the blastocyst stage. The haploid blastomeres of the 2-cell stage were fused electronically, resulting in that the fused embryos showed a significantly higher rate of blastocysts compared to non-fused haploid embryos (55% vs. 37%). Further, the embryonic stem cells (ESCs) derived from the fused embryos were confirmed to be diploid. The rate of ESC establishment in fused embryos was significantly higher compared to non-fused ones. Based on the results, we concluded that the electronic fusion of haploid embryos could be efficient to generate homozygous ESCs.

연초의 약배양 및 종간교배에 의한 반수체 배가계통의 특성 (Characterization of Androgenic and Gynogenic Doubled Haploids from Inbred Cultivars)

  • 금완수;이승철;김달웅
    • 한국연초학회지
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    • 제10권1호
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    • pp.21-26
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    • 1988
  • Ten anther-derived doubled haploid (ADH) and 10 maternally-derived doubled haploid fines (MDH) from NC 2326 were evaluated in the field to compare their performance. Both anther-derived doubled haploid and maternally-derived doubled haploid lines were agronomically inferior to selfed parental variety for many parameters, but the degree of inferiority of the MDH lines were not significantly different from the selfed parental variety for any character investigated. The inferiority of the ADH lines were, however, significantly different from the selfed parental variety for yield, leaf length and width , and days to flower.

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연초(Nicotiana tabacum cv BY4) 이배체 및 반수체 식물의 캘러스로부터 식물체 재생 관련 효소의 변화 (Enzyme Activity in Plant Regeneration from Diploid and Haploid Calli of Nicotiana tabacum cv BY4)

  • 오승철;소웅영;조덕이;양덕춘
    • 식물조직배양학회지
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    • 제21권6호
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    • pp.333-339
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    • 1994
  • 연초의 약배양에 의해서 반수체를 유기하고 형성된 반수체와 이배체의 캘러스로부터 식물체 분화과정에서 일어나는 생리적 차이점을 비교하고자 효소의 활성도와 페놀화합물의 함량을 비교 조사하였다. 반수체의 형성은 화아가 중간정도된 크기를 이용하여 IAA가 1.0 mg/L kietin이 0.5mg/L 조합 처리되고 활성탄이 3 mg/L 함유된 배지에서 가장 양호하였다. 캘러스의 유도는 이배체 및 반수체 공히 2,4-D의 농도가 0.5 mg/L 첨가된 배지에서 양호하였으며, 캘러스로부터 재분화는 BAP 농도 2.0 mg/L에서 효과적이었다. 재분화시 변화되는 효소의 활성도와 페놀화합물의 함량은 처리된 BAP의 농도와 사용한 식물체에 따라 차이가 있었다. Peroxidase의 활성도는 이배체 및 반수체 공히 BAP 농도가 2.0 mg/L일때 가장 높았으며, catalase의 활성도는 BAP의 농도가 1 mg/L일때 가장 높은 경향을 보였다. IAA oxidase 및 catalase의 활성도는 이배체에 비해 반수체에서 더 높은 경향을 보였으나 peroxidase의 활성도는 이배체가 더 높은 경향을 나타내었다.

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벼의 Doubled-haploid 집단육성과 SSR 마커를 이용한 유전자 지도작성 (Development of Doubled-haploid Population and Construction of Genetic Map Using SSR Markers in Rice)

  • 김경민;남우일;권용삼;손재근
    • Journal of Plant Biotechnology
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    • 제31권3호
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    • pp.179-184
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    • 2004
  • 본 연구에서는 1998년부터 2003년 하계까지 약배양 기법 및 콜히친 처리를 이용하여 개발한 '삼강벼/낙동벼' DH(doubled-haploid) 183계통의 주요 농업 형질을 조사$.$분석하였다. DH 집단의 주요 농업형질을 조사한바 초장, 간장, 수장, 삼절간장, 수수 및 출수일수는 양적형질의 특징인 넓은 범위의 변이폭, 연속적인 빈도 분포양상 및 양친을 초월하는 초월분리 현상을 보였다. SSR 마커를 이용한 유전자지도의 작성에는 양친에 다형성을 나타내는 136개의 마커를 사용하였다. 작성된 유전자 지도는 전체 길이가 1,909cM이었으며, 마커간 평균길이는 14 cM을 나타내었다.

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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    • 제31권2호
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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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Cytological and Morphological Characterization of Anther Derived Plants from Sweet Pepper (Capsicum annuum L.) cv. 'Special'

  • Shrestha, Surendra Lal;Luitel, Binod Prasad;Lee, Taek Jong;Kang, Won Hee
    • 한국육종학회지
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    • 제42권5호
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    • pp.431-438
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    • 2010
  • Anthers of sweet pepper $F_1$ cultivar 'Special' were cultured on Dumas De Vaulx (C medium), supplemented with $0.1mgL^{-1}$ 2, 4-D and $0.1mg{\cdot}L^{-1}$ kinetin with 3% sucrose, and 0.32% phytagel. The calluses obtained were further sub-cultured on Murashige and Skoog (MS) medium without growth regulators for regeneration. Regenerated plantlets were grown in plastic pots under plastic house and characterized their cytological and morphological characters in spring, 2008. Twenty percent plantlets were identified as haploid plants after chromosome and ploidy analysis. Haploid plants contained 12 chromosomes, high stomatal density with small stomatal length as compared to diploid plants. Stomatal length in haploids was 23.3% smaller than diploids. Haploid plants were characterized as small leaf and petiole size, poor vigor, thin stem and short plant height, short internodes and small flower buds, fruit size and fruit weight as compared to diploid plants and most of the haploid fruits were seedless. SP55, SP62, SP68, SP72 and SP77 are found high yielding double haploids with high total soluble content (8.6, 8.7, 9.2, 9.1 and $9.8^{\circ}Brix$, respectively) and desirable fruit shape, and recommended them to exploit as inbred lines for heterosis breeding.