• Title/Summary/Keyword: Embryo growth

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Plant Regeneration through Somatic Embryogenesis from Cotyledon of Herbaceous Peony (Paeonia lactiflora Pall.) (芍藥(Paeonia lactiflora Pall.)의 子葉組織으로부터 體細胞胚發생을 통한 식물체 獲得)

  • 신종희;손재근;김경민;박소득;김규원
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.291-294
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    • 1997
  • The experiments were carried out to determine the optimum conditions for the direct embrogenesis from the cotyledon derived zygotic embryo culture of Paeonia lactiflora Pall. Different peony tissues derived from zygotic embryos were cultured on MS medium with and without 2,4-D. Somatic embryos were formed from the cotyledons cultured on the medium without 2,4-D. The somatic embryogenesis from cotyledons was promoted in the growth regulator-free MS medium containing 1.65~3.3 g/L $NH_4NO_3$ and 30~40 g/L sucrose. The maximum frequency (80.0%) of somatic embryo formation was obtained from the cotyledons excised from zygotic embryos that cultured on MS medium containing 3.3 g/L $NH_4NO_3$. Epicotyl and roots were elongated from a somatic embryo by adding 0.3 mg/L GA$_3$ in the medium or the cold treatment at 4$^{\circ}C$ more than three weeks at 4$^{\circ}C$.

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High frequency plant regeneration from zygotic-embryo-derived embryogenic cell suspension cultures of watershield (Brasenia schreberi)

  • Oh, Myung Jin;Na, Hye Ryun;Choi, Hong-Keun;Liu, Jang Ryol;Kim, Suk Weon
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.87-92
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    • 2008
  • An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when cultured on halfstrength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly with increasing concentrations of 2,4-D up to $3mg\;l^{-1}$, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryoderived white friable callus were established using half-strength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of water-shield, which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could be useful for the mass propagation and transformation of selected elite lines.

Influence of Seed Dressing with Captan wp. on the Dehiscence of Panax ginseng Seeds (CAPTAN분의소독이 인삼종자의 개갑에 미치는 영향)

  • Lee, J.C.;Chung, Y.R.;Park, H.;Ohh, S.H.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.28 no.2
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    • pp.262-266
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    • 1983
  • Influence of fungi living on the endocarp surface of depulped seeds of Panax ginseng on the dehiscence was investigated with a fungicidal treatment of the seeds and sterilization of the sand at the beginning of stratification. 1. Isolation frequency of the fungi living on the endocarp surface of de-pulped seed was reduced and hardness of the endocarp did not change significantly in seed treated with a fungicide Captan wp. 50%. A significant negative correlation $(r=-0.984^**)$ was found between the frequency of fungi isolation and the hardness of the endocarp. 2. Increase of water content in the seed treated with the fungicide was delayed 20days compared to the untreated. 3. Growth of the embryo and dehiscence of the seed was suppressed by the fungicide treatment. The length of the embryo was inversely proportional to the hardness of the seed. It is suggested that the fungi facilitate the softening of the endocarp thereby enhancing the supply of oxygen and water necessary for the embryo development, therefore, accelerate the growing of embryo and cause the dehiscence.

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The embryological studies on the interspecific hybrid of ginseng plant (Panax ginseng x P. Quiuquefolium) with special references to the seed abortion (인삼의 종간잡종 Panax ginseng x P Quinquefoilium의 발생학적 연구 특히 결실불능의 원인에 관하여)

  • Jong-Kyu Hwang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.5 no.1
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    • pp.69-86
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    • 1969
  • On the growing of the interspecific hybrid ginseng plant, the phenomena of hybrid vigoures are observed in the root, stem, and leaf, but it can not produce seeds favorably since the ovary is abortive in most cases in interspecific hybrid plants. The present investigation was undertaken in an attempt to elucidate the embryological dses of the seed failure in the interspecific hybrid of ginseng (Panax Ginseng ${\times}$ P. Quinque folium). And the results obtained may be summarized as follows. 1). The vegetative growth of the interspecific hybrid ginseng plant is normal or rather vigorous, but the generative growth is extremely obstructed. 2). Even though the generative growth is interrupted the normal development of ovary tissue of flower can be shown until the stage prior to meiosis. 3). The division of the male gameto-genetic cell and the female gameto-genetic cell are exceedingly irregular and some of them are constricted prior to meiosis. 4). At meiosis in the microspore mother cell of the interspecific hybrid, abnormal division is observed in that the univalent chromosome and chromosome bridge occure. And in most cases, metaphasic configuration is principally presented as 23 II+2I, though rarely 22II+4I is also found. 5). Through the process of microspore and pollen formation of F1, the various developmental phases occur even in an anther loclus. 6). Macro, micro and empty pollen grains occur and the functional pollen is very rare. 7). After the megaspore mother cell stage, the rate of ovule development is, on the whole, delayed but the ovary wall enlargement is nearly normal. 8). Degenerating phenomena of ovules occur from the megaspore mother cell stage to 8-nucleate embryo sac stage, and their beginning time of constricting shape is variously different. 9). The megaspore arrangement in the parent is principally of the linear type, though rarely the intermediate type is also observed, whereas various types, viz, linear, intermediate, Tshape, and I shape can be observed in hybrid. 10). After meiosis, three or five megaspore are some times counted. 11). Charazal end megaspore is generally functional in the parents, whereas, in F1, very rarely one of the center megaspores (the second of the third megaspore) grows as an embryo sac mother cell. 12). In accordance with the extent of irregularity or abnormality in meiosis, division of embryo sac nuclei and embryo sac formation cause more nucellus tissue to remain within th, embryo sac. 13). Even if one reached the stage of embryo sac formation, the embryo sac nuclei are always precarious and they can not be disposed to theil proper, respective position. 14). Within the embryo sac, which is lacking the endospermcell, the 4-celled proembryo, linear arrangement, is observed. 15). Through the above respects, the cause of sterile or seed failure of interspecific hybrid would be presumably as follows, By interspecific crossing gene reassortments takes place and the gene system influences the metabolism by the interference of certain enzyme as media. In the F1 plant, the quantity and quality of chemicals produced by the enzyme system and reaction system are entirely different from the case of the parents. Generally, in order to grow, form, and develop naw parts it is necessary to change the materials and energy with reasonable balance, whereas in the F1 plant the metabolic process becomes abnormal or irregular because of the breakdown of the balancing. Thus the changing of the gene-reaction system causes the alteration of the environmental condition of the gameto-genetic cells in the anther and ovule; the produced chemicals cause changes of oxidatio-reduction potential, PH value, protein denaturation and the polarity, etc. Then, the abnormal tissue growing in the ovule and emdryo sac, inhibition of normal development and storage of some chemicals, especially inhibitor, finally lead to sterility or seed failure. Inconclusion, we may presume that the first cause of sterile or seed abortion in interspecific hybrids is the gene reassortment, and the second is the irregularity of the metabolic system, storage of chemicals, especially inhibitor, the growth of abnormal tissue and the change of the polarity etc, and they finally lead to sexual defect, sterility and seed failure.

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Growth Curve of the Gestational Structures by Ultrasonography Examination in Small Pet Dogs (소형 애완견에서 초음파 검사에 의한 임신 구조물의 성장 곡선)

  • Oh, Ki-Seok;Kim, Bang-Sil;Park, Sang-Guk;Park, Chul-Ho;Kim, Jae-Hong;Mun, Byeong-Gwon;Kim, Hee-Su;Lee, Ju-Hwan;Park, In-Chul;Kim, Jong-Taek;Suh, Guk-Hyun;Son, Chang-Ho
    • Journal of Embryo Transfer
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    • v.23 no.3
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    • pp.147-153
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    • 2008
  • Serial ultrasonographic examinations were daily performed from 15 days after ovulation until parturition to determine the growth curve of gestational structures in pregnant Maltese, Yorkshire terrier, Shih-tzu, and Miniature Schnauzer bitches, respectively. Gestational age was timed from the day of ovulation (day 0), which was estimated to occur when plasma progesterone concentration was first increased above 4.0 ng/ml. The inner chorionic cavity diameter were significantly and linearly relative to gestational age especially days 20 to 40, and the fetal head diameter were significantly and linearly relative to gestational age especial1y day 40 to parturition. These results indicate that inner chorionic cavity diameter were the most accurate for estimating gestational age before day 38 of gestation and the fetal head diameter were after day 38 of gestation.

Studies on the Germination of Korean Ginseng (Panax ginseng C.A.Meyer) Seed II. Influences of Temperature and Seed Treatment on Embryo Growth and Germination (인삼 종자의 발아에 관한 연구 II. 온도 및 종자처리가 배생장 및 발아에 미치는 영향)

  • 원준연;조재성;김현호
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.33 no.1
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    • pp.59-63
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    • 1988
  • Freshly harvested and depulped Korean ginseng seeds were subjected to the seed treatment of removing endocarp plus surface sterilization with sodium hypocloride, surface sterilization only, and nontreated control. These seeds were stratified at temperatures of 5$^{\circ}$, 10$^{\circ}$, 15$^{\circ}$ and 20$^{\circ}C$ for 20, 40, 60, 80 and 100 days. Embryo growth of the ginseng seeds of which endocarp was removed was most rapid in each stratification temperature and that of sterilized seeds was slower than unsterilized seeds after 80 days stratification at 15$^{\circ}$ and 20$^{\circ}C$. About 15$^{\circ}C$ was an optimal stratification temperature for embry growth in ginseng seeds. Chilling treatment at 5$^{\circ}C$ for 100 days was needed for better germination of dehisced ginseng seeds. An optimal germination temperature for the ginseng seed following chilling treatment was about 15$^{\circ}C$.

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Anti-Mullerian Hormone Serum Concentrations in Prenatal and Postnatal Period in Murine

  • Kim, Dae Young
    • Journal of Embryo Transfer
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    • v.28 no.2
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    • pp.149-155
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    • 2013
  • Mullerian inhibiting substance (MIS) is a member of the TGF-${\beta}$ (transforming growth factor-${\beta}$) family whose members play key roles in development, suppression of tumour growth, and feedback control of the pituitary-gonadal hormone axis. MIS is expressed in a highly tissue-specific manner in which it is restricted to male Sertoli cells and female granulose cells. The serum levels of MIS in prenatal and postnatal ICR mice were measured using the enzyme-linked immuno-solvent assay (ELISA) using the MIS/AMH antibody. Mice were grouped by age: the significant periods were at the onset of development. During sex organ differentiation, no remarkable difference between female and male foetus MIS serum levels (both<0.1 ng/ml) was observed. However, MIS serum levels in pregnant mice markedly changed (4.5~12.2 ng/ml). After birth, postnatal female and male mice serum MIS levels changed considerably (male: <0.1~138.5 ng/ml, female: 5.3~103.4 ng/ml), and the changing phase were diametrically opposed (male: decreasing, female: fluctuating). These findings suggest that MIS may have strong associations with not only develop-ment but also puberty. For further studies, establishing the standard MIS serum levels is of importance. Our study provides the basic information for the study of MIS interactions with reproductive organ disability, cancer, and the effect of other hormone or menopause. We hypothesise that if MIS is regularly injected into middle-age women, meno-pause will be delayed. We detected that serum MIS concentration curves change with age. The changing phase is different between males and females, and this difference is significant after birth. Moreover, MIS mRNA is expressed during the developmental period (prenatal) and also in the postnatal period. This finding indicates that MIS may play a significant role in the developmental stage and in growth after birth.

Establishment and Characterization of the Fibroblast Line from Silkie Bantam

  • Li, L.F.;Guan, W.J.;Li, H.;Bai, X.J.;Ma, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.492-499
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    • 2009
  • A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

Effect of Epidermal Growth Factor(EGF) on Early Embryonic Development in Mouse (Epidermal Growth Factor(EGF)가 생쥐 초기배아의 발생에 미치는 영향)

  • Byun, Hye-Kyung;Lee, Ho-Joon;Kim, Sung-Rye;Kim, Hae-Kwon;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.22 no.2
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    • pp.163-170
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    • 1995
  • Growth factors (GFs) produced by the embryo or by the maternal reproductive tract have been reported to regulate the embryonic development and differentiation. Among GFs, EGF as a mitogen plays a role in mitosis and functional differentiation of trophectoderm cells in mouse. The present study was carried out to investigate the effect of EGF on development of mouse embryos and to localize EGF in the mouse oocytes and embryos, which has been reported to be detected in the reproductive tract in mammals. To investigate the effect of EGF on the development of the embryo, mouse 2-cell embryos were cultured to blastocysts stage in Ham's F10 medium, treated with EGF(10-50 ng/ml) for 72 hrs. Immunocytochemistry was performed from oocyte to blastocyst stage with anti-EGF and anti-Mouse IgG, in order to determine the stage which EGF would be expressed in mouse. Exogenous EGF (more than 10 ng/ml) in the culture medium improved the developmental and hatching rates in the mouse embryos. As a result of immunocytochemistry, the embryonic EGF was expressed after the late 4-cell stage. EGF is thought to enhance preimplantation embryonic development and hatching. Exogenous EGF in the culture medium is thought to activate EGF receptor in the late 4-cell embryos and to enhance blastulation and hatching in mouse embryos. It is concluded that EGF enhances the developmental and hatching rates in the mouse embryos.

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Analysis of Coat Color Changes and Hormone Levels in Korean Brindle Cattle (칡소의 모색 발현과 호르몬 변화와의 상관관계 분석 연구)

  • Lee, Hae-Lee;Park, Jae-Hee;Kim, Yong-Su;Kim, Jong Gug
    • Journal of Embryo Transfer
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    • v.30 no.1
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    • pp.65-71
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    • 2015
  • The aim of this study was to determine the relationship between the coat color appearance of Korean brindle cattle and the changes of relevant hormone levels that may affect the hair pigmentation during different stages of growth and maturation. In mature cattle, levels of both ACTH and DHEA in Korean brindle cattle with brown color were significantly higher than those with black color (p<0.05). Levels of ${\alpha}$-MSH in Korean brindle cattle with whole brindle ($${\geq_-}50%$$) color were significantly higher than those with brown color (p<0.05). In calves of Korean brindle cattle at 2 to 6 months, the concentration of estradiol was significantly higher in calves with whole brindle color than those with part brindle color (p<0.05), when the coat color was confirmed. After 6 month of coat color confirmation, levels of testosterone and ACTH increased in calves with part brindle color and were significantly higher than those with whole brindle color (p<0.05). In calves of Korean brindle cattle at 1 or 2 months, there were no significant differences in hormone levels of estradiol, ACTH, DHEA and ${\alpha}$-MSH between the calves with brindle color and brown color, except estradiol before brindle color appearance. Changes of relevant hormone levels at different stage of growth and maturation may affect the pigmentation of coat during the development of cattle. In addition to the current study correlating the different coat colors with relevant hormone levels, investigation of the coat color associated genes expressed in Korean brindle cattle may further clarify the mechanisms of coat color changes during their development.