• Title/Summary/Keyword: Somatic embryos

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Effect of carbon sources on somatic embryogenesis and cotyledon number variations in carrot (Daucus carota L.)

  • Young Jin Lee;Kyu Seog Hwang;Pil Son Choi
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.89-95
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    • 2023
  • In order to investigate the effect of carbon sources on somatic embryogenesis and cotyledon number variations in carrot, embryogenic callus were cultured in the medium supplemented with various concentrations of sucroseor glucose, and with combination of 2% sucrose and various concentrations of mannitol or sorbitol. The maximum number of somatic embryos formed per flask (1,555.70) was obtained in the medium supplemented with 2% sucrose rather than glucose alone or a combination of mannitol or sorbitol and 2% sucrose, and the number of somatic embryos was decreased with the increasing of mannitol or sorbitol concentration. The frequencies of somatic embryos with two cotyledons were 35.14% for sucrose, 19.88% for glucose, 32.55% for mannitol + 2% sucrose, and 38.59% for sorbitol + 2% sucrose, respectively, and the frequencies of abnormal somatic embryos having 3 or more cotyledons were 64.86% for sucrose, 80.12% for glucose, 67.44% for mannitol + 2% sucrose, and 61.41% for sorbitol + 2% sucrose, respectively. Particularly, the frequency of somatic embryos with two cotyledons (59.16%) was the highest in the 2% sucrose medium compared to the frequency of abnormal somatic embryogenesis with three or more cotyledons, and the frequency gradually decreased with increasing concentration of glucose, mannitol or sorbitol. According to these results, it was found that the ratio of abnormal somatic embryo was higher than the normal somatic embryo in carrot, and was shown that somatic embryogenesis and the cotyledon number was affected by the concentrations of sucrose, glucose as carbon source, and mannitol and sorbitol as osmotic agents in culture medium.

In Vitro Germination of Encapsulated Somatic Embryos of Angelica Tree(Aralia elata Seem.) (두릅나무 피복체세포의 기내발아)

  • 박철호
    • Korean Journal of Plant Resources
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    • v.7 no.2
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    • pp.133-135
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    • 1994
  • Germination rate of encapsulated somatic embryos shelved significant differences under different concentrations of AgN03. The highest germination rate of 81.2% was found on MS medium withouthormones mixed with 10 mg/1 of AgN03. In vitro vermiculite planted with encapsulated embryostreated with 10 mg/1 of AgN03 induced 24.7% germination rate, and vermiculite planted with encap-sulated embryos treated with 40 mg/1 or 80 mg/1 of AgNO, induced no germination at all.

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Origin and Development of Single- and Poly-embryos formed Directly on Excised Cotyledons of Ginseng Zygotic Embryos

  • Yang Deok Chun;Choi Kwang Tae
    • Journal of Ginseng Research
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    • v.23 no.2 s.54
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    • pp.74-80
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    • 1999
  • Excised cotyledon segments of ginseng zygotic embryos cultured on MS basal medium without growth regulators produced somatic embryos near the basal excised portion at a high frequency. The frequency of somatic embryo formation on the segments declined along with advancing zygotic embryo maturity. In immature cotyledons, all the cells of the epidermis and subepidermis were smaller and more densely cytoplasmic than those in mature cotyledons, and from which multiple cells participated in embryogenic division to form somatic embryos with multiple cotyledons and fasciated radicles (poly-embryos). But in germinating cotyledons, only the epidermal cells were densely cytoplasmic and singularly competent to develop into somatic embryos resulting in single-embryos with closed radicles. This result means that the origin and development of somatic embryos is determined according to whether the cells participating in embryonic division are in a single state or a massive state relative to cotyledon maturity.

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Mass Production of Eleutherococcus senticosus Plants through in vitro Cell Culture (세포 배양을 통한 가시오갈피(Eleutherococcus senticosus) 묘목의 대량 생산 시스템 개발)

  • Han, Jeong-Yeon;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.167-172
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    • 2003
  • Immature zygotic embryos of Eleutherococcus senticosus seeds matured rapidly within one month when the seeds comprising zygotic embryos were pieced to small size and cultured on 1/2 MS medium. Frequency of somatic embryos formation was declined rapidly when the zygotic embryos germinated and grew to plantlets. Embryogenic cells were induced by consecutive subculture of somatic embryos on MS medium with 1.0mg/L2,4-D. After heart-shaped somatic embryos were induced by suspension culture, these embryos were plated onto petri dish to support maturation of embryos. Germination of embryos occurred on medium with 5mg/L GA$_3$and transferred to culture bowl to stimulate the further growth. Frequency of soil survival of plantlets was influenced by soil mixture (perlite and peatmoss). The suitable combination of perlite and peatmoss was 1:5, and the soil survival rate was 78% after 4 months. The soil transferred plantlets were over-wintered in field condition after defoliation. New year sprouting of plants was achieved successfully and they grew to adult plants. These results indicate that the systematic procecure of plant production in E. senticosus for micro propagation.

Plant Regeneration from Somatic Embryo with Structural Diversity from Leaf Explant Culture of Ostericum koreanum Kitagawa (강활(Ostericum koreanum Kitagawa)의 잎절편체 배양으로 부터 발생된 여러가지 형태의 체세포배를 통한 식물체 재생)

  • 조덕이;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.51-56
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    • 1998
  • This study was carried out in order to establish plant regeneration via somatic embryogenesis from leaf explant of Ostericum koreanum Kitagawa and to elucidate the effects of NAA and cytokinins (kinetin, BA) on the abnormalities of somatic embryo and the relationship between thecotyledon numberand germinability. Calli were formed on leaf explants cultured on MS agar medium supplemented with various concentrations (0, 0.1, 0.5, 1, 2 mg/L) of NAA and cytokinins. The calli were white, watery and soft, became browning during cultures. Somatic embryos were formed from pale yellowish calli derived browning calli. High frequency somatic embryos were observed on MS medium containing 1 mg/L NAA and 0.1 mg/L BA after 60 days of culture. The mature somatic embryos germinated into plantlets without subculture after 2 weeks. The frequency of normal somatic embryo with two cotyledons was 39.8%. On the other hand, cotyledonary abnormalities of somatic embryos were observed at considerable frequency: 33.6% of somatic embryo with one cotyledon, 15.3% cotyledons with three, 8.2% four cotyledons and 3.1% jar shaped cotyledon. Germination frequency of somatic embryos with two cotyledons was 97.4%, and that of the embryos with abnormal cotyledon was almost similar to that of embryos with two cotyledons, except jar shaped somatic embryos (33.3%).

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High Frequency of Plant Regeneration through Cyclic Secondary Somatic Embryogenesis in Panax ginseng

  • Kim, Yu-Jin;Lee, Ok-Ran;Kim, Kyung-Tack;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.36 no.4
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    • pp.442-448
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    • 2012
  • Somatic embryogenesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology such as medicinally important plants. Single embryos develop into normal plantlets with shoots and roots. Therefore, direct single embryogenesis derived from single cells is highly important for normal plant regeneration. Here we demonstrate that the cyclic secondary somatic embryogenesis in Panax ginseng Meyer is a permanent source of embryogenic material that can be used for genetic manipulations. Secondary somatic embryos were originated directly from the primary somatic embryos on hormone-free Murashige and Skoog medium, and proliferated further in a cyclic manner. EM medium (one third of modified MS medium [MS medium containing half amount of NH4NO3 and KNO3] with 2% to 3% sucrose) favored further development of proliferated secondary somatic embryos into plantlets with root system. The plantlets developed into plants with well-developed taproots in half-strength Schenk and Hildebrandt basal medium supplemented with 0.5% activated charcoal.

Cotyledon Structure and Germinability of Somatic Embryos Formed from Inflorescence Explants of Cnidium officinale M. (천궁 (Cnidium officinale M.)의 화기절편으로부터 형성된 체세포배의 자엽구조와 발아)

  • 조덕이;이은경;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.137-142
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    • 2000
  • Calli were induced from the explants of infloresence, petiole and leaf blade of Cnidium officinale on MS medium with 2.4-D, while embryogenic callus was induced only from inflorescence explants. Somatic embryos of 78 per explant were formed during subculture of the explants on medium without 2.4-D after culture on medium with 2 mg/L 2.4-D. Cotyledonary variation, cup-shaped cotyledon of 49% and other abnormal cotyledons of 13.5 % was observed on the somatic embryos. However this variation could be overcomed by the addition of activated charcoal onto culture medium. Somatic embryos at cotyledonary stage germinated on MS basal medium but the germination rate was very poor, blow 50 %. Somatic embryos on the medium with activated charcoal showed improved germinaton.

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Studies on the Induction of Transformation and Multiplication in Orchid Plants I. Formation of Somatic Embryos and Regeneration from Immature Seeds of Bletilla striata (난과식물의 형질전환 유도 및 다량증식에 관한 연구 I. 자란 (Bletilla striata)의 미성숙 종자로부터 체세포배 형성 및 식물체 재분화)

  • 이정석
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.271-276
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    • 1990
  • Our study was carried out for plant regeneration via somatic embryogenesis from immature seeds of Bletilla striata. The highest frequency of embryogenic callus formation was obtained from the immature seeds (at 150 days after pollination) cultured on Hyponex and VW medium supplemented with 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg/l kinetin under the dark condition. Multiple somatic embryos were induced when embryogenic callus was transferred to VW medium without growth regulators under continued illumination. Somatic embryos were observed histologically with scanning electron microscopy. Regeneration of Bletilla striata was obtained from somatic embryos with a well-defined scutellum and coleoptile as well as with one or more shoot primordia and root primordia. We think that these methods for orchid multiplication must be useful to access clonal propagation of orchids.

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Structural Characteristics of Shoot Apex Developed from Somatic Embryos of Daucus carota L. (당근의 체세포배로부터 발생한 경정부의 구조적 특성)

  • 홍성식;소웅영
    • Korean Journal of Plant Resources
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    • v.12 no.2
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    • pp.133-138
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    • 1999
  • The structure of shoot apex in zygotic and somatic embryos of Daucus carota L. cv. Hongshim was observed by using SEM and longitudinal sections. Shoot apex of zygotic embryo was of an inverted boat shape, and these of two, three and four cotyledon somatic embryos were of an inverted boat shape, a pyramid shape and a convex diamond shape, respectively. In zygotic embryo shoot apex is consisted of small cells which are arranged in layers (tunica) and show corpus in central region. In somatic embryos shoot apices are consisted of somewhat large cells which are arranged in irregularly or slight regularly.

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Ascorbic acid increases demethylation in somatic cell nuclear transfer embryos of the pig (Sus scrofa)

  • Zhao, Minghui;Hur, Tai-Young;No, Jingu;Nam, Yoonseok;Kim, Hyeunkyu;Im, Gi-Sun;Lee, Seunghoon
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.944-949
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    • 2017
  • Objective: Investigated the effect and mechanism of ascorbic acid on the development of porcine embryos produced by somatic cell nuclear transfer (SCNT). Methods: Porcine embryos were produced by SCNT and cultured in the presence or absence of ascorbic acid. Ten-eleven translocation 3 (TET3) in oocytes was knocked down by siRNA injection. After ascorbic acid treatment, reprogramming genes were analyzed by realtime reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, relative 5-methylcytosine and 5-hydroxymethylcytosine content in pronucleus were detected by realtime PCR. Results: Ascorbic acid significantly increased the development of porcine embryos produced by SCNT. After SCNT, transcript levels of reprogramming genes, Pou5f1, Sox2, and Klf were significantly increased in blastocysts. Furthermore, ascorbic acid reduced 5-methylcytosine content in pronuclear embryos compared with the control group. Knock down of TET3 in porcine oocytes significantly prevents the demethylation of somatic cell nucleus after SCNT, even if in the presence of ascorbic acid. Conclusion: Ascorbic acid enhanced the development of porcine SCNT embryos via the increased TET3 mediated demethylation of somatic nucleus.