• 제목/요약/키워드: Embryogenesis

검색결과 550건 처리시간 0.031초

In Vitro Production of Indian Citrs Ringspot Virus-Free Plants of Kinnow Mandarin (Citrus nobilis Lour X C. deliciosa Tenora) by Ovule Culture

  • Singh B.;Sharma S.;Rani G.;Zaidi A.A.;Hallan V.;Nagpal A.;Virk G.S.
    • Journal of Plant Biotechnology
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    • 제7권4호
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    • pp.259-265
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    • 2005
  • Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.

Expression of Gas6 Receptors, Tyro3, Axl, and Mertk, in Oocytes and Embryos and Effects of Mertk RNAi on the Oocyte Maturation

  • Kim, Kyeoung-Hwa;Lee, Sang-Eun;Lee, Kyung-Ah
    • 한국발생생물학회지:발생과생식
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    • 제16권3호
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    • pp.195-204
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    • 2012
  • Recently, we reported growth arrest-specific gene 6 (Gas6) as a new maternal effect gene (MEG), that expressed in the oocytes but functioned principally during embryogenesis. Gas6 RNAi-treated oocytes developed to metaphase II (MII) stage but they have affected M-phase promoting factor (MPF) activity and incurred abnormal pronuclear (PN) formation during fertilization. Gas6 is a ligand of TAM family members (Tyro3, Axl and Mertk) of receptor tyrosine kinase (RTK). Purpose of the present study was to evaluate the expression of Tyro3, Axl and Mertk transcripts in oocytes and early embryos. Expression of Gas6 and Mertk mRNA was detectable in oocytes and follicular cells, while Tyro3 and Axl mRNA was expressed only in follicular cells. Expression of Mertk mRNA was relatively constant during oocytes maturation and embryogenesis, but the other receptors, Tyro3 and Axl, were not expressed in oocytes and PN stage of embryos at all. Knockdown of Mertk mRNA and protein by using sequence-specific Mertk double strand RNA (dsRNA) did not affect oocytes maturation. In this case, however, contrary to the ligand Gas6 RNA interference (RNAi), MPF activity had not been changed by Mertk RNAi. Therefore, we concluded that the Gas6-Mertk signaling is not directly related to the oocyte maturation. It is still required to study further regarding the function of Mertk as the receptor of Gas6 during preimplantational early embryogenesis.

High frequency plant regeneration system for Nymphoides coreana via somatic embryogenesis from zygotic embryo-derived embryogenic cell suspension cultures

  • Oh, Myung-Jin;Na, Hye-Ryun;Choi, Hong-Keun;Liu, Jang Ryol;Kim, Suk-Weon
    • Plant Biotechnology Reports
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    • 제4권2호
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    • pp.125-128
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    • 2010
  • Culture conditions were established for high frequency plant regeneration via somatic embryogenesis from cell suspension cultures of Nymphoides coreana. Zygotic embryos formed pale-yellow globular structures and calluses at a frequency of 85.6% when cultured on half-strength Murashige and Skoog (MS) medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. However, the frequency of pale-yellow globular structures and white callus formation decreased slightly with an increasing concentration of 2,4-D up to 10 $mg\;l^{-1}$ with the frequency rate falling to 16.7%. Cell suspension cultures were established from zygotic embryo-derived calluses using half-strength MS medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. Upon plating onto half-strength MS basal medium, over 92.3% of cell aggregates gave rise to numerous somatic embryos and developed into plantlets. Regenerated plantlets were successfully transplanted into potting soil and achieved full growth to an adult plant in a growth chamber. The high frequency plant regeneration system for Nymphoides coreana established in this study will be useful for genetic manipulation and cryopreservation of this species.

Copper, Zinc-Superoxide Dismutase (Cu/Zn SOD) Gene During Embryogenesis of Bombyx mori: Molecular Cloning, Characterization and Expression

  • Hong, Sun-Mee;Kang, Seok-Woo;Goo, Tae-Won;Kim, Nam-Soon;Lee, Jin-Sung;Nho, Si-Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • 제13권1호
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    • pp.23-30
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    • 2006
  • BmCu/Zn SOD was isolated from early embryo of Bombyx mori using microarray analysis. The BmCu/Zn SOD gene was observed during the early embryonic stage with the strongest signal found at the unfertilizaion, fertilization and blastoderm stages. The BmCu/Zn SOD gene encodes a protein of 154 amino acids with a calculated Mr of 15 kDa. The deduced amino acid sequence of BmCu/Zn SOD indicated that the residues that form on the Cu/Zn binding site are conserved and that the sequence is a 60% identity to that of M. domestica. In a phylogenetic tree, Bm SOD was also close to Drosophila SODs rather than other insect SODs. The BmCu/Zn SOD gene exists as a single copy in the genome. Transcripts of BmCu/Zn SOD cDNA were identified by northern blot analysis. The expression of the BmCu/Zn SOD gene was observed weakly in most of larvae, pre-pupae, pupae and adult tissues. Also, the BmCu/Zn SOD gene was observed in early embryonic stage. Although the roles of SODs remains to be further elucidated, the high expression of BmCu/Zn SOD gene at before 24 h post fertilization suggests that this gene is of general importance during early embryogenesis in the Bombyx mod.

가시오갈피의 수집종과 배양조직에 따른 체세포배발생 및 재분화 식물체의 순화 (Effect of Genotype and Explant on Somatic Embryogenesis and Acclimatization of Acanthopanax senticosus)

  • 이성호;유창연
    • 한국약용작물학회지
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    • 제10권3호
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    • pp.217-221
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    • 2002
  • 가시오갈피의 꽃봉오리 엽병, 줄기, 줄기마디, 뿌리, 꽃대, 잎 등 조직으로 부터 캘러스가 유기 되었으며 캘러스 유기재료로는 꽃봉오리, 엽병, 줄기, 줄기 마디, 뿌리 등이 적합하였다. 캘러스 유기율은 한국산 가시오갈피가 일본산과 러시아산보나 높게 나타났다. 캘러스유기에는 2.4-D와 TDZ조합처리가 효과적이었으며 한국가시오갈피의 엽병, 줄기, 액아, 뿌리절편으로부터 형성된 캘러스에서 배발생 캘러스가 형성되었으며 줄기에서 형성된 캘러스로부터 한국, 일본, 러시아산 가시오갈피 모두에서 배발생캘러스가 형성되었다. 엽병에서 형성된 캘러스로부터 배발생캘러스의 유기율이 가장 높았으며 2.4-D 1mg/l 처리가 가장 효과적이었다. 액체배지에서의 체세포배의 생장에는 생장조절물질을 첨가하지 않은 MS기본배지가 적합하였다. 기내에서 잎과 뿌리 분화가 이루어지고 건실하게 자란 식물체를 온실에 옮겨 순화하였을 경우 99.1%의 생존율을 보였다.

Conservation of cis-Regulatory Element Controlling Timely Translation in the 3'-UTR of Selected Mammalian Maternal Transcripts

  • Lee, Hyun-Joo;Lim, Yoon-Ki;Chang, Sang-Ho;Min, Kwan-Sik;Han, Ching-Tack;Hwang, Sue-Yun
    • Genomics & Informatics
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    • 제5권4호
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    • pp.174-178
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    • 2007
  • The earliest stages of mammalian embryogenesis are governed by the activity of maternally inherited transcripts and proteins. Cytoplasmic polyadenylation of selected maternal mRNA has been reported to be a major control mechanism of delayed translation during preimplantation embryogenesis in mice. The presence of cis-elements required for cytoplasmic polyadenylation (e.g., CPE) can serve as a useful tag in the screening of maternal genes partaking in key functions in the transcriptionally dormant egg and early embryo. However, due to its relative simplicity, UA-rich sequences satisfying the canonical rule of known CPE consensus sequences are often found in the 3'-UTR of maternal transcripts that do not actually undergo cytoplasmic polyadenylation. In this study, we developed a method to confirm the validity of candidate CPE sequences in a given gene by a multiplex comparison of 3'-UTR sequences between mammalian homologs. We found that genes undergoing cytoplasmic polyadenylation tend to create a conserved block around the CPE, while CPE-like sequences in the 3'-UTR of genes lacking cytoplasmic polyadenylation do not exhibit such conservation between species. Through this cross-species comparison, we also identified an alternative CPE in the 3'-UTR of tissue-type plasminogen activator (tPA), which is more likely to serve as a functional element. We suggest that verification of CPEs based on sequence conservation can provide a convenient tool for mass screening of factors governing the earliest processes of mammalian embryogenesis.

말똥성게 (Hemicentrotus pulcherrimus)의 수정 및 배 발생률을 이용한 해양배출 폐기물의 독성평가 (Toxicity Assessment of Ocean Dumping Wastes Using Fertilization and Embryo Development Rates in the Sea Urchin (Hemicentrotus pulcherrimus))

  • 황운기;류향미;김성길;박종수;안경호
    • Environmental Analysis Health and Toxicology
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    • 제25권1호
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    • pp.11-18
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    • 2010
  • Toxicity assessment of ocean dumping wastes (dye waste, urban sewage, food waste) were examined in the fertilization and embryo development rates of the Sea Urchin, Hemicentrotus pulcherrimus. Spawning was induced by injecting 1 mL of 0.5 M KCl into coelomic cavity. Males released white or cream-colored sperms and females released yellow or orange-colored eggs. Experiments were began within 30 min the collection of both gametes. The fertilization and embryo development rates test were performed for 10 min and 64 h after fertilization, respectively. The fertilization and embryo development rates in the control condition (not including ocean dumping wastes sludge elutriate) were greater than 90%, but suddenly decreased with increasing of ocean dumping waste sludge elutriate concentrations. The fertilization and normal embryogenesis rates were significantly inhibited in all waste sludge elutriate from dye waste ($EC_{50}$=4.37; $EC_{50}$=1.76), urban sewage ($EC_{50}$=5.79; $EC_{50}$=2.00) and food waste ($EC_{50}$=7.68; $EC_{50}$=2.16), respectively. The NOEC (<3.13) and LOEC (3.13) of fertiliztion and normal embryogenesis rates very similar in all waste sludge elutriate. These results suggest that biological assay using the fertilization and embryo development rates of H. pulcherrimus are very useful test method for the ecological toxicity assessment of ocean dumping wastes.