• Title/Summary/Keyword: Zygotic embryo

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Direct somatic embryogenesis, plant regeneration and genetic transformation of Panax ginseng

  • Park, Yong-Eui;Yang, Deok-Chun;Park, Kwang-Tae;Soh, Woong-Young;Hiroshi Sano
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.85-89
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    • 1999
  • Somatic embryogendesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology. This paper describes the direct somatic embryogenesis from zygotic embryos of Panax ginseng is reversely related to normal axis growth of zygotic embryos by the experiment of various chemical treatments. Under the normal growth condition, the apical tips of embryo axis produced an agar-diffusible substance, which suppressed somatic embryo development from cotyledons. Although the cells of zygotic embryos were released from the restraint of embryo axis, various factors were still involved for somatic embryo development. Electron microscopic observation revealed that the ultrastructure of cells of cotyledon epidermis markedly changed before initiation of embryonic cell division, probably indicating reprogramming events into the cells embryogenically determined state. Polar accumulation of endogenous auxin or cell-cell isolation by plasmolysis pre-treatment is the strong inducer for the somatic embryo development. The cells for the process of somatic embryogenesis might be determined by the physiological conditions fo explants and medium compositions. Direct somatic embryos from cotyledons fo ginseng were originated eithrer from single or multiple cells. The different cellular origin of somatic embryos was originated either from single or multiple cell. The different cellular origin of somatic embryos was depended on various developmental stages of cotyledons. Immature meristematic cotyledons produced multiple cell-derived somatic embryos, which developed into multiple embryos. While fully mature cotyledons produced single cell-derived single embryos with independent state. Plasmolysis pretreatment of cotyledons strongly enhanced single cell-derived somatic embryogenesis. Single embryos were converted into normal plantlets with shoot and roots, while multiple embryos were converted into only multiple shoots. GA3 or a chilling treatment was prerequisite for germination and plant conversion. Low concentration of ammonium ion in medium was necessary for balanced growth of root and shoot of plantlets. Therefore, using above procedures, successful plant regeneration of ginseng was accomplished through direct single embryogenesis, which makes it possible to produce genetically transformed ginseng efficently.

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Formation of Gametophytes and Development of Zygotic Embryo in Dicentra spectabilis (금낭화(Dicentra spectabilis)의 배우체 형성 및 접합자배의 발달)

  • Sim Ock-Kyeong;Lee Kang-Seop;Kim Ee-Yup;Eun Jong-Seon
    • Korean Journal of Plant Resources
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    • v.18 no.2
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    • pp.302-308
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    • 2005
  • This study was carried out to obtain the basic informations on the characteristics of gametophytes formation and embryo developement in Dicentra spectabilis. Microspore mother cells developed from archesporial cells, start meiosis when flower bud length reaches around 1 mm, formed tetrahedral type tetrad. The 4 microspores were separated. They were developed to male gametophytes, respectively. Megaspore mother cells were observed when flower bud length was $4{\sim}5\;mm$. The developemental type of megaspore was polygonum and embryo sac was amphitropous. Three large and distinctive antipodals did not degenerated and remained after embryo sac was developed. When the male and female gametophytes was fully developed, the length of stamen and style was very similar or stamen was shorter about 0.5 mm than that of style. This result indicates that self-fertilization can be occurred in this species. After fertilization, developing zygotic embryos showed various stages of development from globular to cotyledonary embryos, and zygotic embryo in seed scattering time seemed to have an early cotyledonary stage.

Cryopreservation of Hevea brasiliensis zygotic embryos by vitrification and encapsulation-dehydration

  • Nakkanong, Korakot;Nualsri, Charassri
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.333-339
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    • 2018
  • The mature zygotic embryos of the Hevea brasiliensis were cryopreserved through the use of the vitrification and encapsulation/dehydration techniques. In all the experiments, the zygotic embryos were pre-cultured for three days in the MS medium supplemented with 0.3 M sucrose before they were used for the cryopreservation technique. In the vitrification procedure, the effect of the plant vitrification solutions (PVS2 and PVS3) and exposure time were studied. The highest survival rate (88.87%) and regrowth (66.33%) were achieved when the precultured zygotic embryos were incubated in a loading solution for 20 minutes at $0^{\circ}C$. They were subsequently exposed to PVS2 for 120 minutes at $0^{\circ}C$ and plunged directly into liquid nitrogen. Cryopreservation by the encapsulation-dehydration method was successfully done by leaving the encapsulated zygotic embryos in a laminar flow for 4 hours prior to plunging into a LN. The survival rate and regrowth of the encapsulated zygotic embryos were 37.50% and 27.98%, respectively. The cryopreserved zygotic embryos were able to develop into whole plants.

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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    • v.4 no.2
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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.

Plant Regeneration and Somatic Embryogenesis from Zygotic Embryo-derived Callus of Native Prunus yedoensis in Mt. Halla (한라산 자생 왕벚나무 접합자배 유래의 캘러스로부터 체세포배 형성과 식물체 재분화)

  • 고정군;박영철;양두영;김응식;오문유;고석찬
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.345-349
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    • 1997
  • Somatic embryos were induced through embryogenic callus derived from immature zygotic embryo culture of native Prunus yedoensis in Mt. Halla and regenerated into plantlets successfully. Embryogenic callus was induced most effectively on MS medium with 1.0 mg/L 2, 4-D and 0.1 mg/L BAP at an efficiency of approximately 60% using 45 day-old zygotic embryos after full blooming. Globular somatic embryos were induced from embryogenic callus on MS medium with 1.0 mg/L 2, 4-D and 0.1 mg/L BAP and these globular embryos developed to heart-shaped and cotyledonary embryos on hormone-free MS medium. Normal somatic embryos germinated 49% on 1/2 MS medium and the plants regenerated from the somatic embryos were morphologically normal.

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Cryopreservation of Zygotic Embryos of Herbaceous Peony (Paeonia lactiflora Pall.) by Encapsulation-Dehydration

  • Kim Hyun-Mi;Shin Jong-Hee;Sohn Jea-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.354-357
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    • 2004
  • A simplified technique which cryoprotects zygotic embryos by encapsulation-dehydration was developed for the germplasm conservation of herbaceous peony (Paeonia lactiflora Pall.). The highest survival rate $(85\%)$ was obtained from embryos treated by encapsulation-dehydration. The zygotic embryos were precultured on MS medium containing 0.3mg/L $GA_3$ for 1 day. The precultured embryos were encapsulated in $3\%$ (w/v) alginate beads and immersed for 1 h in MS medium containing 2 M glycerol and 0.5 M sucrose. The encapsulated embryos were dehydrated for 5h by air drying prior to direct immersion in liquid nitrogen. This encapsulation-dehydration method appears to be a promising technique for germplasm cryopreservation of a herbaceous peony.

Plant Regeneration through Somatic Embryogenesis from Cotyledon of Herbaceous Peony (Paeonia lactiflora Pall.) (芍藥(Paeonia lactiflora Pall.)의 子葉組織으로부터 體細胞胚發생을 통한 식물체 獲得)

  • 신종희;손재근;김경민;박소득;김규원
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.291-294
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    • 1997
  • The experiments were carried out to determine the optimum conditions for the direct embrogenesis from the cotyledon derived zygotic embryo culture of Paeonia lactiflora Pall. Different peony tissues derived from zygotic embryos were cultured on MS medium with and without 2,4-D. Somatic embryos were formed from the cotyledons cultured on the medium without 2,4-D. The somatic embryogenesis from cotyledons was promoted in the growth regulator-free MS medium containing 1.65~3.3 g/L $NH_4NO_3$ and 30~40 g/L sucrose. The maximum frequency (80.0%) of somatic embryo formation was obtained from the cotyledons excised from zygotic embryos that cultured on MS medium containing 3.3 g/L $NH_4NO_3$. Epicotyl and roots were elongated from a somatic embryo by adding 0.3 mg/L GA$_3$ in the medium or the cold treatment at 4$^{\circ}C$ more than three weeks at 4$^{\circ}C$.

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Somatic Embryogenesis and Plant Regeneration from Immature Zygotic Embryo Culture of Wasabia japonica Matsum. (고추냉이의 미숙배배양으로부터 체세포배 발생과 식물체 재분화)

  • 은종선;고정애;김영선;김명준
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.207-211
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    • 1995
  • Immature zygotic embryos from immature seeds of Wasabia japonica (cv Dalma) were isolated and cultured on modified MS medium supplemented with 2,4-D, IAA, and BA. Immature zygotic embryos were classified into torpedo shape and cotyledon stage. The highest rates of callus formation were obtained of 1.0mg/L IAA(torpedo stage, 90.0%)and 1.0mg/L 2,4D plus 0.1mg/L BA(cotyledany stage,84.3%). Somatic embryos after 60 days of culture. These numerous somatic embryo could be seperated and subcultured on the same media for further propagation. After 90 days of culture, most somatic embryos were developed well organized embryos which were able to produce into whole plants.

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Somatic Embryogenesis in Immature Zygotic Embryo Cultures of Korean Soybean (Glycine max L.) Culitivars and Effect of 2,4-Dichlorophenoxyacetic Acid on Somatic Embryo Morphology (한국 품종 대두(Glycine max L) 미숙배로부터 체세포배발생과 배지의 2,4-Dichlorophenoxyacetic Acid 농도가 체세포배의 형태에 미치는 영향)

  • Pil S Choi;Yoong Y. SOH;Duck Y.Choi;ang R. LIU
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.7-14
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    • 1994
  • Immature zygotic embryos of five Korean soybean cultivars cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) produced somatic embryos without forming an intervening callus. The highest frequency (up to 90%) of somatic embryo formation was obtained when zygotic embryos were cultlued on medium containing 1 to 2 mga 2, 0-D in four cultivars. However the frequency was highly variable to the cultivars. Transversely sliced primary somatic embryo halves were also capable of forming secondary embryos at frequencies of up to 70% when cultured on medium containing 0.1 to 1 mg/L 2,4-D. Somatic embryos formed on zygotic embryos cultured on medium containing 0.1 to 0.2 mg/L 2,4-D had two cotyledons more frequently than one horn-type cotyledon and those on medium containing 0.5 to 4mg/L 2,4-LD had a horm-type cotyledon at a prominently higher freequency. However somatic embryos on medium containing 10mg/L or higher concentrations of 2,4-D were usually shunted at the globular stage even after transfer to medium containing lower concentrations of 2,4-D or other growth regulators. non somatic embryos with one or two cotyledons or a hem-type cotyledon were transferred to medium containing $GA_3$, those with two cotyledons converted to plantlets at a higher frequency (25%) than the others.

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