• 제목/요약/키워드: immature embryo

검색결과 206건 처리시간 0.047초

Development of a Protocol for Somatic Embryogenesis of Cnidium officinale M akino

  • Hui Yeong Jeong;Ji Ah Kim
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2021년도 춘계학술대회
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    • pp.51-51
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    • 2021
  • This study was conducted to develop a somatic embryogenesis protocol for the Cnidium officinale Makino difficult to seed propagation. The immature flowers were used as explants. The concentration of a 2,4-D 1.0mg/L was found to be optimal concentration for induction of embryogenic callus and somatic embryos. Addition of 0.3mg/L, 0.5mg/L and 1.0mg/L to the embryo germination medium promoted somatic embryo germination. Among four concentrations, GA3 1.0mg/L were superior to others. Shoots were transferred to hormone-free MS medium after 2 months of culture in the dark. We obtained an optimized protocol for the regeneration of C. officinale.

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Esterase isozyme patterns in developing plant regeneration from calli of citrus junos Sieb.

  • Lee, Hyun-Hwa;Lee, Sook-Young;Park, Min-Hee;Jang, Hyun-kyu;Kim, Hong-Sub
    • Plant Resources
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    • 제2권1호
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    • pp.1-9
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    • 1999
  • The callus from the hypocotyl region of immature embryo of Citrus junos Sieb. was efficiently induced in the $\frac{1}{2}$ MS medium containing 45uM BA after 8 weeks culture. The callus was developed into the two callus type, embryogenic callus and nonembryogenic callus, which can be distinguished by visual examination depending on color and appearence. In vitro regeneration of callus established efficiently in the hormone-free MS medium from the embryogenic callus. In order to investigate the physiological changes depending on the developmental stage of embryo, the embryo was formed in the MS medium. The embryogenic and nonembryogenic callus, and the various stages of the somatic embryo were examimed the changes of esterase activity, and their isozyme patterns as well. The protein content and esterase activities was gradually increased on the developmental stages of embryo. Total protein pattern were different by the SDS-PAGE and were appeared strong band of 23 KD in the torpedo stage. The pattern of the esterase isozyme was exhibited a difference between embryogenic callus and nonembryogenic callus. It was appered pI 6.0, 8.0, 8.2 in the embryogenic callus. Also the new band of pI 4.75 was appeared in the cotyledon. These results suggest that the changes of esterase activities and isozyme patterns are importent factor in the differentiation and development of citrus.

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Fertilization and pregnancy potential of immature oocytes from stimulated intracytoplasmic sperm injection cycles

  • Shin, Seung Bi;Cho, Jae Won;Lee, Sun-Hee;Yang, Kwang Moon;Lim, Chun Kyu;Lee, Hyoung-Song
    • Clinical and Experimental Reproductive Medicine
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    • 제40권1호
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    • pp.7-11
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    • 2013
  • Objective: We evaluated the fertilization potential of immature oocytes obtained from controlled ovarian hyperstimulation cycles of patients undergoing ICSI. Methods: We retrospectively analyzed 463 ICSI cycles containing at least one immature oocyte at oocyte denudation. ICSI was performed on mature oocytes at oocyte denudation (metaphase-II [MII] oocytes) and the oocytes that extruded the first polar body between oocyte denudation and ICSI (MI-MII oocytes). Fertilization and early embryonic development were compared between MII and MI-MII oocytes. To investigate the pregnancy potential of MI-MII oocytes, the pregnancy outcome was analyzed in 24 ICSI cycles containing only immature oocytes at retrieval. Results: The fertilization rate of MI-MII oocytes (37.0%) was significantly lower than that of MII oocytes (72.3%). The rates of delayed embryos and damaged embryos did not significantly differ. Eighty-one immature oocytes were retrieved in 24 cycles that retrieved only immature oocytes and 61 (75.3%) of them were in the MI stage. ICSI was performed on 36 oocytes (59.0%) that extruded the first polar body before ICSI and nine MI-MII oocytes (25.0%) were fertilized. Embryo transfers were performed in five cycles. Pregnancy was observed in one cycle, but it ended in biochemical pregnancy. Conclusion: In ICSI cycles, oocytes that extruded the first polar body between denudation and ICSI can be used as a source of oocytes for sperm injection. However, their fertilization and pregnancy potential are lower than that of mature oocytes. Therefore, ovarian stimulation should be performed carefully for mature oocytes obtained at retrieval, especially in cycles with a small number of retrieved oocytes.

대두 체세포배에서 전형성층 분화와 경단분열조직의 발달 (Procambium differentiation and shoot apical meristem development in somatic embryos of soybean (Glycine max L.))

  • 최필선;권석윤
    • Journal of Plant Biotechnology
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    • 제40권1호
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    • pp.55-58
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    • 2013
  • 대두 미숙배 배양으로부터 체세포배를 얻기 위해 1 mg/L 2,4-D를 첨가한 MS배지에서 6 - 8주 동안 배양한 결과 2개의 자엽을 갖는 정상체세포배가 14%, 1개의 자엽을 갖는 비정상 체세포배가 37%, 합생자엽을 갖는 비정상 체세포배가 43% 그리고 생장이 멈춘 구형기 체세포배가 6%로 얻어졌다. 체세포배의 전형층은 하배축에서 원형모양으로 분화되어 자엽 절로 이어진 후 1개의 자엽은 1개 전형성층이, 2개의 자엽은 2개의 전형성층이 각 자엽으로 발달하였고, 합생 자엽의 경우 하배축에서 형성된 원형의 전형성층이 자엽까지 연결되어 있었다. 한편 2개의 자엽을 갖는 체세포배의 경우는 tunica-corpus층을 이룬 전형적인 유경조직을 이루고 있었으나 1개 또는 합생 자엽을 갖는 체세포배는 tunica-corpus층이 없고, dome모양이 없는 흔적만 남은 유경조직을 이루고 있었다. 이러한 결과는 체세포배에서 전형성층의 분화는 자엽의 형성과 유경조직의 발달에 밀접한 관계가 있음을 시사한다.

인간난자의 체외수정에서 난포기의 Estradiol 반응도와 양상이 임신율과의 판계에 관한 연구 (The Correlation Between the Preovulatory $E_2$ Pattern and Pregnancy Rate after in Vitro Fertilization-Embryo Transfer)

  • 이상훈;최훈;김정구;문신용;이진용;장윤석
    • Clinical and Experimental Reproductive Medicine
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    • 제14권2호
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    • pp.109-118
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    • 1987
  • Ninety-one patients with irreparable tubal disease underwent in Vitro Fertilization-Embryo Transfer (IVF-ET) in Seoul National University Hospital. Ovulation was stimulated in 104 cycles by human menopausal gonadotropin (HMG) or follicular stimulating hormone (FSH)/human chorionic gonadotropin (HCG). The patients were classified as high (>900 pg/ml), intermediate (400-900 pg/ml), or low (<400pg/ml) responder according to preovulatory $E_2$ response and four $E_2$ patterns were found. The overall pregnancy rate per cycle in this consecutive series was 11.5% (n=12). The number of preovulatory oocytes per cycle was higher significantly in intermediate and high responder group than in low responder group (P<0.01), While the number of immature oocytes per cycle significantly higher in low responder group than high and intermediate responder group (P<0.01). The pregnancy rate in each responder group was not signigicant. According to the $E_2$ pattern of response, there was no significant difference in number of the immature and preovulatory oocytes recovery per cycle. There was a apparently direct relationship between the preovulatory $E_2$ pattern and pre gnancy rate was noted.

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A Comparative Study on the Parthenogenetic Development of Pig Oocytes Cultured in North Carolina State University-23 and Porcine Zygote Medium-3

  • Lee, Joo-Hyeong;Hyun, Sang-Hwan;Lee, Eun-Song
    • 한국수정란이식학회지
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    • 제27권2호
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    • pp.121-126
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    • 2012
  • The objective of this study was to examine the effect of in vitro culture media on embryonic development of in vitro-matured (IVM) oocytes after parthenogenetic activation (PA) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 22~26 h. IVM oocytes were activated by electric pulses and cultured in porcine zygote medium-3 (PZM-3) and North Carolina State University-23 supplemented with essential and non-essential amino acids (NCSU-23aa). These media were further modified by supplementing 2.77 mM myo-inositol, 0.34 mM trisodium citrate, and $10{\mu}M$ ${\beta}$-mercaptoethanol (designated as mPZM-3 and mNCSU-23aa, respectively). Culture of PA embryos in mPZM-3 significantly increased development to the blastocyst stage than culture in NCSU-23aa (36.2% vs. 24.8%, p<0.05). Modified PZM-3 showed a significantly higher blastocyst formation than NCSU-23aa in both groups of embryos that were activated at 44 h and 48 h of IVM (51.0% vs. 35.5% and 49.0% vs. 34.2% in oocytes activated at 44 h and 48 h of IVM, respectively). Irrespective of the follicle diameter where oocytes were collected, embryonic development to the blastocyst stage was increased (p<0.05) by the culture in mPZM-3 compared to culture in NCSU-23aa (25.9% vs. 34.2% and 32.9% vs. 44.8% in embryos derived from small and medium size follicles, respectively). Our results demonstrated that culture media had significant effect on preimplantation development PA embryos and that mPZM-3 was superior to mNCSU-23 in supporting development to the blastocyst stage in pigs. This beneficial effect of mPZM-3 on embryonic development was not impaired by other factors such as time of oocyte activation and origin of immature oocytes (small and medium size follicles).