• Title/Summary/Keyword: Embryo size

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Development of complete Culture System for Quail Embryos and Its Application for Embryo Manipulation

  • Ono, T.
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.155-163
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    • 2001
  • Gene and cell transfer technique will serve as a powerful tool for the genetic improvement of the poultry and to yield useful products. For avian transgenesis, Japanese quail may serve as an excellent animal model because of its small body size and fast growth rate. Recent progress was described on the manipulation of quail embryos such as the introduction of foreign genes and cells, and the subsequent culturing of the manipulated embryos yielding hatchlings. Intraspecific donor-derived offspring have been available in quail, however, further investigation will be required to obtain interspecific offspring with the aim of rescuing endangered species. Trans genesis will also be useful for improving the profitability and quality of poultry stocks and for developing stocks with novel uses. Considerable progress should soon be made toward the production of transgenic poultry. The key feature of the procedure described here is that embryos are initially taken out from the shell for ease of manipulation and then placed back in culture in addition to various operations midway during culture.

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Effect of Sperm Concentration on Fertility and Litter Size in Deep Frozen Boar Semen (돼지 동결정액 중의 정자농도가 수태율 및 산자수에 미치는 영향)

  • 김학규;정행기;이수헌;이광원;김인철;최진성;지병천;김경남;박창식
    • Journal of Embryo Transfer
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    • v.11 no.1
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    • pp.81-83
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    • 1996
  • This study was carried out to investigate the effect of sperm concentration of 5ml maxi-straw on farrowing rate and number of pigs born alive per litter in deep frozen boar semen. We did not find out the effect of sperm concentration on post-thaw sperm motility and NAR acrosome. However, farrowing rate and number of pigs born alive per litter of 7. 5 x 10˚ /5ml and 10.0 x 10˚ /5m1 sperm concentrations were higher than those of 5. 0 /10˚ /5ml sperm concentration.

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Description of the External Genitalia and Uterus of a 24-month-old Freemartin Hanwoo

  • Kim, Ui-Hyung;Kang, Sung-Sik;Chung, Ki-Yong;Yang, Boh-Suk;Cho, Sang-Rae
    • Journal of Embryo Transfer
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    • v.33 no.1
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    • pp.13-16
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    • 2018
  • We observed the external genitalia and uterus of a 24-month-old freemartin Hanwoo. The vulva was smaller than observed in a normal female Hanwoo, while the clitoris was larger in the freemartin. The angle between the external genitalia and the perineum also varied. Upon internal genital examination, the uterus of the freemartin was a thin tube approximately 18 cm in size and had not differentiated into a normal uterus and uterine horns.

Factors Affecting In-Vitro Maturation in Porcine Oocytes (돼지난자의 체외성숙에 영향을 미치는 요인)

  • 박춘근
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.179-191
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    • 1996
  • In-vitro culture has provided new inforrnation on mechanisms of oocytes rnaturation and results obtained in vitro have led to new questions. In porcine, follicular and oocyte size have the crucial importance for the oocytes maturation. The addition of hormones to the culture medium was found to accelerate and facilitate meiotic maturation. The presence of some factors in serum trigger the resumption of meiosis and support the maturation of oocytes in vitro. The maturation rate of porcine oocytes was also increased by supplementation of porcine follicular fluid to the culture medium. The growth factors can stimulate nuclear maturation and enhances cytoplasnic maturation of oocytes by interaction with gonadotropins. The maturation-promoting factor brings about GVBD and the subsequent maturational events in oocytes. However, cAMP can block the spontaneous meiotic maturation of oocytes in culture. The understanding of these influences is a prerequisite to enhancing in vitro maturation of porcine oocytes.

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Efficient plant regeneration from immature embryo cultures of Jatropha curcas, a biodiesel plant

  • Varshney, Alok;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.139-148
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    • 2010
  • Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.

Embryo lethality and teratogenicity of 2-Bromopropane in the Sprague-Dawley rat (Sprague-Dawley 랫드에서 2-Bromopropane의 배자치사 및 최기형성 효과)

  • Kim, Jong-Choon;Oh, Ki-Seok;Shin, Dong-Ho;Kim, Sung-Ho;Kim, Hyeon-Yeong;Yun, Hyo-In;Jiang, Cheng-Zhe;Heo, Jeong-Doo;Chung, Moon-Koo
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.657-666
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    • 2003
  • The present study was undertaken to evaluate the potential adverse effects of 2-BP on pregnant dams and embryo-fetal development after maternal exposure during the gestational days (GD) 6 through 19 in Sprague-Dawley rats. The test chemical was administered subcutaneously to pregnant rats at dose levels of 0, 375, 750 and 1250 mg/kg/day. During the test period, clinical signs, mortality, body weights and food consumption were examined. All dams were subjected to caesarean section on GD 20 and their fetuses were examined for external, visceral and skeletal abnormalities. At above 750 mg/kg, toxic effects including signs of toxicity, suppressed body weight, decreased gravid uterine weight and reduced food intake were observed in pregnant dams. An increase in the fetal deaths, a decrease in the litter size, a reduction in the fetal body weight and an increase in the incidence of fetal morphological alterations were also found. There were no adverse effects on either pregnant dams or embryo-fetal development at a dose level of 375 mg/kg. These results suggest that a 14-day subcutaneous dose of 2-BP is embryolethal and teratogenic at above 750 mg/kg/day in pregnant rats. In the present experimental condition, the no-observed-adverse-effect level of 2-BP is considered to be 375 mg/kg/day for dams and embryo-fetuses, respectively.

An Optimized Method for the Construction of a DNA Methylome from Small Quantities of Tissue or Purified DNA from Arabidopsis Embryo

  • Yoo, Hyunjin;Park, Kyunghyuk;Lee, Jaehoon;Lee, Seunga;Choi, Yeonhee
    • Molecules and Cells
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    • v.44 no.8
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    • pp.602-612
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    • 2021
  • DNA methylation is an important epigenetic mechanism affecting genome structure, gene regulation, and the silencing of transposable elements. Cell- and tissue-specific methylation patterns are critical for differentiation and development in eukaryotes. Dynamic spatiotemporal methylation data in these cells or tissues is, therefore, of great interest. However, the construction of bisulfite sequencing libraries can be challenging if the starting material is limited or the genome size is small, such as in Arabidopsis. Here, we describe detailed methods for the purification of Arabidopsis embryos at all stages, and the construction of comprehensive bisulfite libraries from small quantities of input. We constructed bisulfite libraries by releasing embryos from intact seeds, using a different approach for each developmental stage, and manually picking single-embryo with microcapillaries. From these libraries, reliable Arabidopsis methylome data were collected allowing, on average, 11-fold coverage of the genome using as few as five globular, heart, and torpedo embryos as raw input material without the need for DNA purification step. On the other hand, purified DNA from as few as eight bending torpedo embryos or a single mature embryo is sufficient for library construction when RNase A is treated before DNA extraction. This method can be broadly applied to cells from different tissues or cells from other model organisms. Methylome construction can be achieved using a minimal amount of input material using our method; thereby, it has the potential to increase our understanding of dynamic spatiotemporal methylation patterns in model organisms.

Development of a Miller Producing Clean White Embryo Rice Using a Vertical Miller (입형정미기를 이용한 청결배아미 제조기 개발)

  • 엄천일;정종훈
    • Journal of Biosystems Engineering
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    • v.29 no.2
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    • pp.121-130
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    • 2004
  • This study was carried out to develop a miller to produce white embryo rice with functional nutrients by improving the conventional vertical miller. The effects of rice moisture content and the shaft revolution speed of the miller on germ(embryo) adherence rate, whiteness, broken rice rate, and cracked rice rate were investigated. Also, the effect of the mesh size of emery stones on the germ adherence rate was investigated. The vertical prototype miller was improved with the increasement of about 42% in producing white embryo rice at proper conditions(shaft revolution speed of 900 rpm, emery stones of mesh #50, processing capacity of 2.3t/h, zero outlet resistance, rice moisture content of 16.2%). The results were as follows: 1. The germ adherence rate of white rice was significantly influenced by the moisture content of brown rice. The germ adherence rate of white rice decreased rapidly with the increase of the moisture content of brown rice. When brown rice with moisture content of 13.2%, 14.5%, 15.2%, 15.4% was milled by the prototype with emery stones of mesh #35 and shaft speed of 900(1,100) rpm, rpm adherence rate of milled rice was 76.2%(70%), 69.2%(66%), 45.9(38%), 13.0(9%), respectively. 2. The whiteness of white rice milled by the prototype with emery stones of mesh #35 and shaft speed of 1,100(900)rpm increased from 27(23) to about 40, respectively, as the moisture content of brown rice increased from 13.2% to 17.2%. 3. The rate of broken rice of white rice milled at 900rpm decreased by 0.6∼1.0% compared with that at 1,100rpm when the moisture content of brown rice was less than 15.2%. 4. The germ adherence rate was increased by 10.3% and 11.0%, respectively when brown rice with moisture content of 16.2% and 15.5% was milled by the prototype miller with shaft speed of 900rpm and emery stones of mesh #50 instead of mesh #35. 5. Considering the germ adherence rate, broken rice rate, and whiteness of milled rice, the proper milling conditions of the prototype miller for producing embryo rice were the moisture content of about 15%, the processing capacity of 2.3t/h and minimum outlet resistance of 0Nㆍm with shaft speed of 900rpm and emery stones of mesh #50.

STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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