• Title/Summary/Keyword: Embryo rescue

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Abnormal Seedlings Emerged during Embryo Rescue and Its Remedy for Seedless Grape Breeding

  • Ji, Wei;Li, Zhiqian;Yao, Wenkong;Gong, Peijie;Wang, Yuejin
    • Horticultural Science & Technology
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    • v.31 no.4
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    • pp.483-489
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    • 2013
  • The abnormal seedlings, a common physiological anomalies, emerged during embryo rescue severely restricted grape breeding. To enhance the efficiency of the seedless grape breeding by reducing the production of abnormal seedlings in the course of embryo rescue, we investigated the effects of genotype, media type, embryo style, pre-chilling on the deformity rate of the abnormal seedlings during embryo rescue. The abnormal seedlings were firstly classified into seven categories based on their morphology. Our results indicated that the emergence of abnormal seedlings was highly dependent on the female parent genotype. Polyembryony was advantageous to diminish the number of abnormal plantlets and the germination rate of embryo was 100%. We also found that pre-chilling treatment could reduce the number of abnormal plantlets and promote the embryo germination. The abnormal plantlets were reduced significantly by the addition of $ZnSO_4$ $10{\mu}mol{\cdot}L^{-1}$ or mashed-banana $500mg{\cdot}L^{-1}$ to either embryo development or germination media. Transferring the abnormal seedlings onto the suitable fresh media in 4 weeks after embryo germination provided an effective way to transform them into normal seedlings.

Embryo Rescue Efficiency Affected by Developmental Stages of Embryo and Medium Composition in Early-Ripening Peach (Prunus persica)

  • Sewon Oh;Byeonghyeon Yun;Se Hee Kim;Sang-Yun Cho;Namhee Jung;Kyung Ran Do;Kang Hee Cho;Hyun Ran Kim
    • Korean Journal of Plant Resources
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    • v.37 no.3
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    • pp.263-269
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    • 2024
  • Embryos of early-ripening peaches could not achieve physiological maturation or undergo abortion before harvest. Embryo rescue is an effective strategy to rescue embryos from early-ripening peaches. Thus, the current study was carried out to determine the appropriate developmental stage and optimal medium composition for embryo rescue in early-ripening peach. Development of open-pollinated 'Yumi' fruit was investigated from 20 to 90 days after full bloom (DAFB) to explore period occurring endocarp hardening. After endocarp hardening, embryo development was observed by light microscopes. Shoot and root meristems were observed at 65 DAFB and embryo size rapidly increased at 75 DAFB. Embryos collected at 75, 80, 85, and 90 DAFB were cultured on four media based on Driver and Kuniyuki (DKW) medium. Germination rate of embryos cultured on four media gradually increased from 75 to 90 DAFB and reached 100% at 90 DAFB. Notably, M3 medium (0.5 DKW supplemented with 6-benzylaminopurine (BAP) 1.0 ㎎/L) displayed the highest germination rate at 75 and 80 DAFB stages. Growth and development of shoot and root were pronounced in plantlet cultured at 90 DAFB stage. While delayed shoot growth was evident in plantlets cultured at 75, 80, and 85 DAFB stages, this retardation could be overcome through the application of growth regulators, particularly in M3 and M4 (0.5 DKW supplemented with BAP 1.0 ㎎/L and indole-3-butyric acid 0.5 ㎎/L) media. Remarkably, roots of plantlet grown in M4 medium exhibited limited elongation. In conclusion, germination rate of embryo and growth of embryo cultured plantlet can be enhanced by collecting seeds from early-ripening 'Yumi' at the 90 DAFB stage and conducting embryo culture using the M3 medium.

Comparison of embryonic competence and clinical outcomes between early and late cumulus cell removal for in vitro fertilization

  • Pongsuthirak, Pallop
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.4
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    • pp.362-367
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    • 2021
  • Objective: The impact of early mechanical removal of cumulus cells on fertilization and embryonic development is not yet precisely known. This study aimed to investigate the effects of early and late cumulus cell removal on fertilization, polyspermy, embryonic development potential, blastocyst development, and clinical outcomes. Methods: A prospective study was conducted of patients who underwent in vitro fertilization between September 2019 and October 2020. Sibling oocytes were randomly allocated after insemination to early cumulus cell removal at 6 hours (group I) and late cumulus cell removal at 16-18 hours (group II). If total fertilization failure (TFF) was determined to have occurred at early cumulus cell removal, rescue intracytoplasmic sperm injection (ICSI) was performed. Fertilization, embryonic development, and pregnancy outcomes were compared. Results: A total of 912 oocytes were assigned to group I (458 oocytes) and group II (454 oocytes). Fertilization, polyspermy, embryo quality, and pregnancy outcomes were not significantly different between both groups. Rescue ICSI enabled fertilization of 79.2% of the TFF oocytes. Conclusion: Early cumulus cell removal at 6 hours had no significant difference in fertilization, polyspermy, embryo development, or obstetric and perinatal outcomes compared to late removal. Early cumulus cell removal combined with early rescue ICSI may have the potential to help couples with TFF.

Effects of mechanical Injury, cotyledon elimination and nutrition for the germination of apple seeds (Malus pulmila var. domestica) (사과 종자 발아에 미치는 기계적 상처, 자엽제거 및 영양분의 영향)

  • Choi, Pil Son;Lee, Young Jin;Yu, Young Beob;Choi, Myung Suk
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.318-322
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    • 2019
  • When dehulled apple seeds (Malus pilmila var. domestica) were induced to germinate, the rate of germination, which was increased by piercing both the radicle and cotyledon, was related to the site of injury. Also, the germination rates of an embryo with ¾ cotyledon (¾ CE), embryo with ½ cotyledon (½ CE), embryo with ¼ cotyledon (¼ CE), and embryo without cotyledon (NCE) gradually decreased compared to an embryo with a normal cotyledon, but it significantly recovered on the agar media containing MS solution and/or 80 mg/L gibberellin (GA3) as nutrition. The results show that the germination of the embryo was inhibited by the partial removal of cotyledon and also demonstrated that the rate of germination could be significantly increased by adding GA3 (80 mg/L) and/or MS medium.

M-RAS Regulate CDH1 Function in Blastomere Compaction during Porcine Embryonic Development

  • Zhou, Dongjie;Niu, Yingjie;Cui, Xiang-Shun
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.12-20
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    • 2020
  • Cell adhesion plays an important role in the differentiation of the morphogenesis and the trophectoderm epithelium of the blastocyst. In the porcine embryo, CDH1 mediated adhesion initiates at compaction before blastocyst formation, regulated post-translationally via protein kinase C and other signaling molecules. Here we focus on muscle RAS oncogene homolog (M-RAS), which is the closest relative to the RAS related proteins and shares most regulatory and effector interactions. To characterize the effects of M-RAS on embryo compaction, we used gain- and loss-of-function strategies in porcine embryos, in which M-RAS gene structure and protein sequence are conserved. We showed that knockdown of M-RAS in zygotes reduced embryo development abilities and CDH1 expression. Moreover, the phosphorylation of ERK was also decreased in M-RAS KD embryos. Overexpression of M-RAS allows M-RAS KD embryos to rescue the embryo compaction and blastocyst formation. Collectively, these results highlight novel conserved and multiple effects of M-RAS during porcine embryo development.

Enhanced Seed Development in the Progeny from the Interspecific Backcross (Fagopyrum esculentum ${\times}$ F. homotropicum) ${\times}$ F. esculentum

  • Shin, Dong-Hoon;Kamal, A H M;Yun, Young-Ho;Bae, Jeong-Sook;Lee, Yun-Sang;Lee, Moon-Soon;Chung, Keun-Yook;Woo, Sun-Hee
    • Korean Journal of Plant Resources
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    • v.22 no.3
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    • pp.209-214
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    • 2009
  • To facilitate the introgression of F. esculentum into the traits of F. homotropicum, several accessions of the hybrids between these two species were pollinated with F. esculentum as the recurrent parent. The embryo in vitro rescue was performed to increase the recovery of backcross progenies. The $F_{2}$ generation was more amenable than $F_{1}$ hybrids to produce backcross progenies. The $F_{1}$ hybrids were backcrossed twice with common buckwheat (pin-type F. esculentum) (recurrent backcrossing). Also, alternate backcrosses with common buckwheat and F. homotropicum (congruity backcrossing) were carried out. Pollen tube growth of BC$F_{1}$ ${\times}$ F. esculentum (thrum) and F. homotropicum ${\times}$ BC$F_{1}$ was the disturbed penetration exceeded for all initial interspecific hybrids, and its requirement was proportionally lower when the common buckwheat was used as the recurrent parent and as the last parent of congruity hybrids. Effects of both common buckwheat and F. homotropicum on seed success rate for hybridization were observed. Growth of hybrid embryos before rescue, regeneration of mature hybrids all increased recurrent and congruity backcrosses and inter-crosses between $F_{1}$ plants and selected fertile plants of the second congruity backcrosses.

Effect of Co culture System with Autologous Cumulus Cells on Embryo Quality and Pregnancy Rates (체외수정시술주기에서 배아와 난구세포의 공배양 효과에 관한 연구)

  • Hur, Eui-Jong;Lee, Won-Ki
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.3
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    • pp.299-304
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    • 1998
  • Despite the rapid development of assisted reproductive technologies (ART) in recent years, implantation rates after replacement of embryos into the uterine cavity remains low. Several techniques such as culture conditions based on formulations of human tubal fluid and various ART techniques as GIFT, ZIFT, TET have been adopted in recent years to improve embryo viability in vitro and implantation rates. Also, coculture of human IVF-derived embryos have been used in an effort to increase the number of viable embryos following IVF and to improve synchrony between the developing embryo and the uterine environment. The aim of this study was to evaluate whether the use of co culture with autologous cumulus cells has a significant beneficial effect on the development of embryos in vitro and its relation to the pregnancy rates in 120 patients with previous failed IVF-ET from September, 1995 to January 1998. We obtained the results from which significant improvement in the quality of viable embryos were observed using a coculture system with autologous cumulus cells, but pregnancy rates in this group of patients did not differ from the rate in the standard IVF group during the same period. Our study shows that a simplified short-term coculture system with autologous cumulus cells may help rescue moderate quality embryos to cleave regularly.

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Application of in vitro ovary culture for cottonwood (Populus deltoides) breeding

  • Kang, Ho-Duck;Lee, Min-Soon
    • Plant Resources
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    • v.7 no.2
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    • pp.87-92
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    • 2004
  • Five different poplar hybrids were tested for rescuing embryo to elongate in vitro plantiets after hybridization. Ovaries and ovules were cultured on Woody Plant Medium (WPM) supplemented with cytokinins, 6-benzylamine (BA) and zeatin. Multiple shoots were initiated from half section of capsule with immature embryos after 21 days from pollination and tiny shoots were formed after the expansion of cotyledons in ovule cultures. Germinating response was better in intraspecific hybrids $(6.53\pm1.66)$ than interspecific crosses $(0.93\pm0.54)$ from half section of capsules on WPM medium. In general, zeatin was better than BA in inducing multiple shoots from isolated ovules. The highest average number $(19.40\pm4.53)$ of shoots was produced from immature ovules of 21 days post-pollination of WPM medium supplemented with 5.0 mg/L zeatin. The highest percentage of germination was 93% from the half section of in vitro ovary cultures. Soil acclimation was successfully conducted in cell tray containing artificially mixed soil with 96% survival rate.

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