• Title/Summary/Keyword: 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.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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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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    • v.5 no.3
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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.

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

  • Jung, Hyo Sun;Kim, Youn Kyoung;Kim, Hyun Chul;Noh, Jae-Koo;Lee, Jeong-Ho;Kim, Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.5
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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.

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

  • 정윤화;금완수
    • Journal of the Korean Society of Tobacco Science
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    • v.16 no.2
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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
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.1
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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.

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

  • 금완수;이승철;김달웅
    • Journal of the Korean Society of Tobacco Science
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    • v.10 no.1
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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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Enzyme Activity in Plant Regeneration from Diploid and Haploid Calli of Nicotiana tabacum cv BY4 (연초(Nicotiana tabacum cv BY4) 이배체 및 반수체 식물의 캘러스로부터 식물체 재생 관련 효소의 변화)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.6
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    • pp.333-339
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    • 1994
  • Enzyme activities and phenolic compound were compared to investigate the physiological characteristics during shoot formation from diploid and haploid of Nicotiana tabacum w BY4. The Nakata medium with 1.0 mg/L IAA, 0.5 mg/L Kinetin and 3 g/L activie carbon was excellent to induce the haploid plants from the middle size anther within 30 days after culture. The MS medium with 0.5 mg/L 2,4-D was good for callus induction from leaf explants of diploid and haploid, and a lot of plane were regenerated from calli on the MS medium supplemented with 2.0 mg/L BAE Activities of peroxidase for both of diploid and haploid plane were the highest at 2.0mg/L BAP Activities of IAA oxidase and catalase of haploid Plants were high or than those of diploid plants. On the other hand, activity of peroxidase of haploid plants were lower than those of diploid plants.

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

  • Kim, Kyung-Min;Nam, Wu-Il;Kwon, Yong-Sham;Sohn, Jae-Keun
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.179-184
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    • 2004
  • A doubled-haploid (DH) population was developed through anther culture of F$_1$ plants obtained from a cross between a japonica cultivar, 'Nagdongbyeo', as male parent and a indica cultivar, 'Samgangbyeo', as female parent. Segregation modes for plant length, culm length, panicle length, third internode length, and days to heading in the DH lines showed nearly normal distribution with wide range of variation. A molecular map with 136 simple sequence repeat (SSR) markers was constructed using the DH population. The total map distance was 1,909 cM and the average interval of marker distance was 14 cM.

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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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
    • Korean Journal of Breeding Science
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    • v.42 no.5
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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.