The aim of this study was to determine the effect of additional alpha-linolenic acid (ALA) supplementation during in vitro maturation (IVM) and culture (IVC) on nucleic maturation and embryo development of pigs. Cumulus-oocyte complexes (COCs) were incubated in IVM medium containing different concentration of ALA (25, 50 and 100 μM) for 44 h. After in vitro maturation, nuclear maturation of oocytes were evaluated by aceto-orcein stain. Mature oocytes with 50 μM ALA were fertilized and cultured in IVC medium with ALA (25, 50 and 100 μM) during early-embryogenesis (48 hours after fertilization). Then, embryos were cultured with 25 μM ALA during early embryogenesis and/or late embryogenesis (120 hours after early-embryogenesis). In results, oocyte maturation were significantly increased by 50 μM ALA treatment groups compared with control groups (p<0.05). Treatment of 25 μM ALA during early-embryogenesis enhanced cleavage rate of embryo compared with other groups (p<0.05), whereas formation and total cell number of blastocyst had no significant difference. Similarly, cleavage rate of embryos were increased by 25 μM ALA supplement during early- or late-embryogenesis than ALA treatment both stage of embryogenesis (p<0.05), but did not influence to blastocyst formation. Interestingly, total cell number of blastocyst were enhanced in ALA treatment group during early-embryogenesis. These findings indicated that ALA supplement enhance the nuclear maturation of oocyte and embryo development, however, excessive ALA could negatively influence. Therefore, we suggest that ALA is used for improvement of in vitro production of mammalian embryo and further study regarding with functional mechanism of ALA is needed.

Early maturing glutinous rice lines with giant embryo were developed using anther culture. Deuraechan, mid-late maturing high-yielding japonica rice variety with resistance against rice stipe virus (RSV), bacterial blight (BB), and lodging, and Chenghyangna ge, early maturing glutinous rice germplasm with giant embryo were used the parents. F2 seeds from the cross between Deuraechan and Chenghyangna ge with glutinous endosperm and giant embryo were selected and propagated to F2 population. In F2 population, anther culture was conducted using the panicles from the early maturing plants. All doubled haploid (DH) lines showed early maturing, glutinous endosperm, and giant embryo phenotype. Through marker-assisted selections to Stvb-i and Xa3, 17 DH lines carrying both resistance genes were selected. Among 17 DH lines, six lines with more embryo size and better agronomic traits were selected and analyzed their characteristics. These lines were early maturing glutinous rice with giant embryo and showed enhanced yield, resistance against RSV and BB, and lodging, compared to previously developed giant embryo rice varieties. But they were vulnerable to preharvest sprouting which is important trait in early maturing rice. According to the texture and rapid viscosity analysis, DH lines were considered to have appropriate properties of cooked brown rice. They showed less hardness, gummniess, chewiness, and setback. Developed DH lines could be useful materials for diversification of cropping system and enhancing the brown rice consumption but the breeding efforts to improve the vulnerability against preharvest sprouting is required to apply for practical variety.

This study monitored the morphological development of embryo, larvae and juvenile yellowtail kingfish, Seriola lalandi, for their aquaculture. The fertilized eggs obtained by natural spawning were spherical shape and buoyant. Fertilized eggs were transparent and had one oil globule in the yolk, with an egg diameter of 1.35 ± 0.04 mm and an oil globule diameter of 0.32 ± 0.02 mm. The fertilized eggs hatched 67–75 h after fertilization in water at 20 ± 0.5°C. The total length (TL) of the hatched larvae was 3.62 ± 0.16 mm. During hatching, the larvae, with their mouth and anus not yet opened. The yolk was completely absorbed 3 days after hatching (DAH), while the TL of post-larvae was 4.72 ± 0.07 mm. At 40 DAH, the juveniles had grown to 30.44 ± 4.07 mm in TL, body depth increased, the body color changed to a black, yellow, and light gray-blue color, and 3–4 vertical stripes appeared. At 45 DAH, the juveniles were 38.67 ± 5.65 mm in TL and 10.10 ± 0.94 mm in body depth. The fish were green with a light orange color, with 7 faint green-brown stripes on the sides of their body. At 87 DAH, the juveniles had grown to 236.11 mm in TL, 217.68 mm in fork length, and 136.5 g in weight. The fish resembled their adult form, with a light yellow-green body color, loss of the pattern on the sides of their body, and a yellow coloration at the tip of the caudal fin.

Abeliophyllum distichum is a monotypic taxon of Oleaceae and endemic to Korea. A comprehensive study on embryogeny and fruit and seed coat ontogeny in Abeliophyllum was carried out via microtome and light microscopy. The fertilization occurs during mid– to late April and embryo matures by early July. The embryo development follows the general fashion from globular embryo – transition embryo – heart shaped embryo – torpedo embryo – walking-stick embryo to mature embryo. The pericarp clearly differentiates into three histological zones: exocarp, mesocarp, and endocarp. The young seed comprises 10-12 cells thick seed coat and the mature seed coat comprises an exotesta, 6-8 mesotesta and an endotesta. Any crystals, phenolic-like compounds, idioblasts, and the sclereids are not found in pericarp as well as seed coat. An overall development confirms Solanade type of embryogenesis in Abeliophyllum. The endocarp becomes more prominent in mature fruit

Environmental conditions during early mammalian embryo development are critical and some adaptational phenomena are observed. However, the mechanisms underlying them remain largely masked. Previously, we reported that AQP5 expression is modified by the environmental condition without losing the developmental potency. In this study, AQP11 was examined instead. To compare expression pattern between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP11 by whole mount immunofluorescence. When the fertilized embryos were developed in the maternal tracts, the level of Aqp11 transcripts was decreased dramatically until 2-cell stage. Its level increased after 2-cell stage and peaked at 4-cell stage, but decreased again dramatically until morula stage. Its transcript level increased again at blastocyst stage. In contrast, the levels of Aqp11 transcript in embryos cultured in vitro were as follows. The patterns of expression were similar but the overall levels were low compared with those of embryos grown in the maternal tracts. AQP11 proteins were localized in submembrane cytoplasm of embryos collected from maternal reproductive tracts. The immune-reactive signals were detected in both trophectoderm and inner cell mass. However, its localization was altered in in vitro culture condition. It was localized mainly in the plasma membrane of the blastocysts contacting with external environment. The present study suggests that early stage embryo can develop successfully by themselves adapting to their environmental condition through modulation of the expression level and localization of specific genes like AQP11.

Adaptive development of early stage embryo is well established and recently it is explored that the mammalian embryos also have adaptive ability to the stressful environment. However, the mechanisms are largely unknown. In this study, to evaluate the possible role of aquaporin in early embryo developmental adaptation, the expression of aquaporin (AQP) 5 gene which is detected during early development were examined by the environmental condition. To compare expression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5 by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantation embryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expression pattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second mid-preimplantation gene activation phase, but the fold changes were modified between in vivo embryos and in vitro embryos. During in vivo development, AQP5 was mainly localized in apical membrane of blastomeres of 4-cell and 8-cell stage embryos, and then it was localized in cytoplasm. However, the main localization area of AQP5 was dramatically shifted after 8-cell stage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

p63은 다양한 상피 조직의 줄기세포와 전구세포에 존재한다는 사실이 잘 알려져 있으나, 치아 형성, 특히 사기질과 뿌리 형성시기에서의 p63 위치느 ㄴ아직 연구해야 할 과제로 남아 있다. 본 연구에서는 p63이 치아 발생 동안 치아상피에 편재하여 나타나는 것을 면역조직화학 기법을 이용하여 확인하였다. p63은 피부, 모낭, 구강점막 그리고 턱밑샘 도관을 포함하는 상피의 바닥층과 바닥위층에 위치하였다. 그러나 치아 부위에서는 치아관의 모든 세포, 사기질기

This study was conducted to determine the characteristics of fertilization process and embryo development of Astragalus membranaceus Bunge (Astragali Radix) to provide basic data needed in its breeding. A. membranaceus showed poor seed setting when self-pollination was induced. When artificial pollination was induced, it showed less than 5% bearing in late August, but more than 13% bearing from the beginning of September 4th. The flower size was about 17.0 mm×4.0 mm and pistils and stamens had the same length of 15.0mm at flowering stage. When self-pollination or cross-pollination was induced, pollen tubes extended to an ovule. While pollen tube was extending to the ovule, reproductive cell split and formed two male generative nuclei and a vegetative nucleus. In the case of self-pollination, fertilized embryo was not observed, but was formed in the case of cross-pollination. A. membranaceus is noted to have zygote self-incompatibility. In the case of cross-pollination, fertilization was observed in 6 to 8 h after pollination, where apical cell derivatives split after fertilization. A spherical pro-embryo was then formed three days after fertilization. The seed attained full shape with a seed coat showing its distinctive contour 15 days after fertilization. Thus, A. membranaceus in Leguminosae family is found to have zygote selfincompatibility although its flower shape is shown to match the self-compatibility plant.

Effects of salinity and standard toxic metals on fertilization and embryo development rates were investigated in the sea urchin, Strongylocentrotus nudus. Spawning was induced by injecting 1㎖ of 0.5 M KCl into the coelomic cavity. The fertilization and embryo development rates were below 20% when salinity was 25 psu or lower, but were above 90% when salinity was between 30 and 35 psu. The fertilization and embryo development rates in the control condition (not including Cu and Cd) were greater than 90%, but decreased with a high negative correlation (r) of 0.89 and 0.91 with the increasing of Cu and Cd concentrations, respectively. These results suggest that salinity concentrations for successful fertilization and normal embryogenesis of S. nudus are between 30 and 35 psu, and the biological assays of fertilization and embryo development rates using S. nudus are useful methods for the ecological toxicity test of marine pollution elements.

본 연구는 금낭화 배우체의 형성과 배 발달특성에 관한 기초 자료로 활용하기 위해 수행되었다. 시원세포로부터 발달된 소포자 모세포는 화뢰의 길이가 1 mm일 때 감수분열을 하여 4면체형(tetrahedral) 4분자가 형성되었다. 4개의 소포자는 분리되어 각각 웅성배우체로 발달하였다. 대포자 모세포는 화뢰의 길이 4~5 mm에서 관찰되었다. 대포자의 발달유형은 정상형(polygonum)이었으며, 배낭의 형태는 굽어있는 곡생배주(amphitropous)였다. 3개의 뚜렷한 반족세포는 배낭이 성숙한 후에도 퇴화하지 않고 남아있었다. 개화 전 자웅배우체는 충분히 성숙하였다. 개화 시에 수술과 암술의 길이는 거의 비슷하거나 수술이 0.5 mm 짧아서 자화수정에 적당한 구조를 가지는 것으로 나타냈다. 수정 후 배는 구형, 심장형을 거쳐 자엽배까지 발달하였으며, 종자 산포시 초기 자엽배를 가지고 있음을 알 수 있었다.

컴퓨터 세포동결기를 이용하여 생쥐 배아를 동결할 때 액체질소 (L)의 분사속도가 해빙 후 배아의 미세구조, 기능 및 발달에 미치는 영향을 알아보고자 하였다. 이를 위해 배아는 동결을 하지 않은 대조군 (control) 및 동결군에서 L의 분사속도에 따라 고속분사군 (120 infusion/min group 1), 저속분사군 (50 infusion/min; group 2)으로 나누었다. ICR 계열의 생쥐의 2 세포기 배아를 사용하였으며, 동결 및 해빙은

이온 비의존적으로 작동하는 포도당 수송체 (glucose transporter 1, Glut1)는 생쥐 배아의 세포막을 경계로 포도당을 수송하는 주요통로이다. 성장인자 가운데 insulin-like growth factor-I (IGF-I)은 생쥐배아에서 포도당의 유입을 증가시키는 것으로 알려져있으나 이러한 효과가 IGF-I 의한 Glut1의 전사조절 효과에 기인한 것인지는 알려져 있지 않다. 본 연구는 포도당과 IGF-I 생쥐의 착상전 배아 발생과 G

Phospholipase C (PLC)는 다양한 세포주에서 세포내 신호전달에 중요한 역할을 한다고 알려져 있으나, 생쥐 난자성숙 과정과 착성전 배아발생 과정에서 PLC의 역할과 발현은 아직 연구된 바 없다. 본 연구에서는 난자성숙과 착상전 배아발생 과정에서 생쥐의 PLC β1과 γ1의 유전자 발현을 조사하기 위하여 한 개의 난자 혹은 배아에서 추출된 total RNA를 사용하여 경쟁적 RT-PCR 방법으로 mRNA를 정량하였다. PL

Festuca속의 토올페스큐와 메도우페스큐, 그리고 Lolium속의 이탈리안라이그라스들 간에 종속간 잡종을 얻기 위하여 기내수분과 기내배양을 실시하였다. 이들 작물들의 자방과 화분을 채취하여 소독을 하고 주두수분, 주두제거 수분, 배주수분의 3가지 방법으로 기내수분을 실시하였다. 그리고 이들의 배발육상태를 MS, N6 , White's의 배지종류와 IAA Kinetln, BA의 생장조절물질의 처리로서 비교하였다. 1. 토올페스큐, 메도우페스큐, 이탈리안라이그라스의 상호 조합간 기내수분에서 44~92%의 수정율을 나타내었다. 이들의 주두수분은 평균 67.8% 주두제거 수분은 평균 89%의 수정율을 보였으나 배주수분은 평균 61 %로 낮았다. 2. 화본과 수정배주에서 배발육을 촉진하는데는 White's 배지가 가장 효율적이었고 식물생장조절물질의 처리는 IAA(10mg/ )와 Kinetin(0.2 mg/ )의 혼합처리가 무처리보다 효과적이었으나 발아율은 극히 저조하였다 3. 기내배양으로 성숙 배주의 배부조직에서 유관책의 형성과 시원체조직의 분화를 확인할 수 있었으며 배주 내부에 후막세포들의 형성이 관찰되었다.

수분후 시기별 배발달 상태와 종자형질의 생장, 개갑 및 발아특성을 연관시켜 조사 연구하여 인삼종자의 채종가능시기와 채종적기를 구명하였던 바 그 결과를 요약하면 다음과 같다. 1. 수분후 열매, 종자 및 배유의 크기는 초기생장이 빨라 20일경에는 성숙한 종자 크기와 큰 차이가 없어 조기채종종자와 완숙종자간에만 유의차가 인정되었으나 종자중은 시기가 경과할 수록 점차 무거워지는 경향이었다. 2. 수분후 20일경에 채종한 종자는 전혀 개갑이 되지 않았으나 30일 후에 채종한 종자는 개갑과 발아가 비교적 양호하여 수분 30일 후면 종자채종이 가능한 시기로 사료되었다. 3. 인삼의 채종적기는 중앙부위의 소화 적심시 수분후 약 50일경의 완숙기에 채종하는 것이 바람직하였다. 4. 배의 발아시기는 수분후 약 10일경으로 추측되었으며 배의 생장속도는 초기단계에서 빠른 경향을 보여 채종가능시기인 수분후 30일경의 배장은 200mu extrmm 이상으로 자엽조직이 분화하기 시작하였다.

팔굉, 노풍, TN1 3품종을 공시하여 20℃ 하에서 인공수분하여 수정과정을 조사하고, 상기 3품종외 이외326, 밀양29, 수원264등 9개품종을 일반포장에서 이앙기를 달리하여 감戮수분열과 등숙비율을 조사한 결과는 단음과 같다. 1 팔굉은 30℃ 하에서는 수분후 1시간30분에서 4시간후에, 20℃ 하에서는 2시간30분에서 4시간30분후에 수정이 된다. 2. 노풍과 TN1은 30℃ 하에서는 2시간에서 5시간30분후에, 20℃ 하에서는 수분후 3시간에서 6시간후에 수정이 된바. 3. 저온인 20℃ 하에서는 수정률이 저하하고, 특히 TN1에서는 극핵과 정핵 또는 난세포와 청핵의 수정이 거의 동시에 되는 것과 어느 한편만 수정이 되는 때도 있다. 4. 조발육에 있어서는 20℃ 하에서는 팔굉이 빠른 것 같고, 30℃ 하에서는TN1이 빠른 것 같다. 5. 팔굉은 이앙기가 늦어도 감수분열에 이상이 없이 정상화분립을 형성하나, TN1은 감수분열에 이상이 있어 불념화분립수가 많아진다.