• Title/Summary/Keyword: Artificial fertilization

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Sex Detection and In Vitro Development of Biopsied Bovine Embryo for LAMP Based Embryo Sexing (LAMP 방법에 의한 소 수정란의 성 판별과 Biopsy에 따른 수정란의 체외발달)

  • Cho S. R.;Choi S. H.;Kim H. J.;Han M. H.;Choe C. Y.;Chung Y. G.;Son D. S.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.169-176
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    • 2005
  • Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

Studies on the Method of Ground Vegetation Establishment of Denuded Forest Land in the Mudstone Region - The Characteristics of Mudstone and Speeded-up Reforestation - (니암지대황폐림지(泥岩地帶荒廢林地)의 지피식생(地被植生) 조성방법(造成方法)에 관(關)한 연구(硏究) - 니암특성((泥岩特性)과 조기녹화(早期綠化) -)

  • Chung, In Koo
    • Journal of Korean Society of Forest Science
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    • v.19 no.1
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    • pp.1-23
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    • 1973
  • The results of ground vegetation experiment conducted at completely denuded forestland in the mudstone region are summerized as follows: On the reaults of soiling quantity the effect of soiling was observed where depth of soiling over 10 cm was practiced, and a plot where treated with 15cm soiling and without fertilizer showed poor growth and it was even worser than the plot where soiling was practiced only 1 cm in thikness but applied adequate amount of fertilizers. The depth of slits between 30cm and 40cm showed no significant difference in the effect. A plot where covered with vegetation bag showed somewhat better results in seed loss and early growth but no differences observed in the fall result over the none covered plot. And then, it is recommendable to have soiling over 10cm in thikness with slit of 30cm and 30cm in depth and to apply 30 gram of fertilizer (22;22:11, 50 gram) per slit. On various soiling materials trial there were no striking differences in the effect of soiling between weathered granite soil, wheathered tuffs soil and weathered mudstone soil. In the treatment with various green materials, a plot treated with straw mat showed a significant difference at 1 percent. The results show that weathered mudstone soil is effective to use as soiling materials and straw mat treatment was better. On forest fertilization trial, in the mudstone region where red and black pine trees already existing at a rate of 2,000-3,000 trees per hectare had applied 110kg of compound fertilizers (9:12:3 and 22:22:11) per hectare basis in terms of plant nutrient. As a result, the difference in effect between the compound fertilizers was not found however the leaf color and leaf length of the fertilizer added plot showed darker and longer at 30 percent over the no fertilizer received plot. Compound fertilizers, 14:37:12 and 9:12:3 were applied to alder trees at a rate of 20 gram and 40 gram per tree in terms of plant nutrient and a remarkable growth accelerantion was observed where 40 grams of plant nutrient applied. The effect difference between the compound fertilizers was not found. On investigation of tree root elongation, forty years old red pine trees showed only 15cm tap root elongation through mudstone while black pine had 23 cm tap root elongation. The total length of supporting root elongtion of red and black pines showed 20 and 13 meters, respectively. The tap roots of Black locusts were not able to elongate through mudstone, however, the supporting roots tended to develop to the underneath of pine tree where some moisture content is available. Black locusts And grown on the residual soil of mudstone normally die between 8 to 10 years. The red pine trees show flat in tree shape while black pine had triangle in the shape. With the results it can be said that in an artificial reforestation in denuded forest land of the mudstone region the adequate slit and enough amount of fertiliizer application must be provided for the succesful performance of the program. On integrated experimental results of 1972. for the establishment of ground vegetation on the completely denuded forest land in mudstone region, soiling could be effectively practiced with weathered mudstone soil and it would not specially necessiate to have either weathered granite or tuffssoil for the soiling. And the soiling depth should be more than 10 cm in thickness. Among green materials used the straw mat proved to be the most effective reatment. Three major factors which enable to establish ground vegetation by the shortest period of time: A. Physical improvement of soil is necessary to breakdown of the horizontal cracks sushas Slit, contour line plot, seeding hole and etc., and soiling with weathered mudstone soil. B. Chemical improvement of soil: is needed sufficient amount of fertilizer application 300~400kg ha, $N+P_2O_5+K_2O$), and increased production of ground covering and expedite resolution of the vegetation (ground vegetation, fallen leaves and twigs). C. Complete establishment of the basic structure for the erosion control (Prevention of surface soil erosion)

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