• Title/Summary/Keyword: secondary somatic embryogenesis

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Effect of Casein on Somatic Embryogenesis and Plant Regeneration in Shoot Apical Meristem Explants of Sweetpotato (Ipomoea batatas L.) (고구마 정단분열조직으로부터 체세포배발생 및 식물체 재분화에 미치는 casein의 영향)

  • Shin, Kong-Sik;Roh, Kyung-Hee;Lee, Yeon-Hee;Park, Young-Whan;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.67-72
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    • 2004
  • An efficient protocol has been developed for rapid mass propagation of sweetpotato from shoot-tips derived embryogenic callus. Optimal embryogenic callus was induced from shoot apical meristem explants on Murashige and Skoog (MS) medium supplemented with 1mg/L 2,4-D. The addition of casein hydrolysate in the media increased the embryogenesis efficiency of sweetpotato. Somatic embryos were easily induced from the embryogenic callus on MS basal medium containing 300-500mg/L casein hydrolysate without phytohormon. Treatment of casein hydrolysate (100∼300mg/L) with 1mg/L 2,4-D also improved the secondary embryonic efficiency from somatic embryos below 2mm in length. Plant regeneration was achieved via somatic embryogenesis and direct organogenesis. Regenerated planlets with well developed shoots and roots on MS basal medium were successfully transferred to soil.

Somatic embryogenesis from the axillary meristems of peanut (Arachis hypogaea L.)

  • Singh, Shweta;Hazra, Sulekha
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.333-340
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    • 2009
  • Developmental anomalies in the plumule meristem of peanut (Arachis hypogaea L.) somatic embryos resulted in poor shoot differentiation and reduced plant recovery. Existing meristems with caulogenic potential have never been tested for embryogenesis in peanut. The present experiment was designed to test the mature zygotic embryo axis derived plumule with three meristems for somatic embryogenesis. Embryogenic masses and embryos developed from the caulogenic meristems in the axils. Exposure of 2 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation temporarily which then regained the ability to form the shoot on withdrawal of 2,4-D. Exposure of 4 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation irreversibly. No shoot formation was noted from the tips in any of the cultures which were in secondary medium with $13.6{\mu}M$ 2,4-D. Development of somatic embryos directly from axillary meristems was confirmed histologically. Conversion frequency of these embryos was 11%. Thus, in this report, we describe a method to obtain somatic embryos from the determined organogenic buds of the axillary meristem, by culturing the nodal explant vertically on embryo induction medium. It also displays the possibility of obtaining both embryogenic and organogenic potential in two parts of the same explant simultaneously. The possibility of extending this approach for genetic transformation in in vivo system through direct DNA delivery or Agrobacterium injection in meristems can also be explored. Using Agrobacterium rhizogenes, we have demonstrated the possibility of gene transfer in the axillary meristems of seed-derived plumule explant.

Plantlet Formation and Somatic Embryogenesis from the penduncle explants of wild Corydalis remota for. peatinata (야생 현호색 (Corydalis remota for. peatinata)의 꽃자루로부터 체세포배 발생과 소식물체형성)

  • 천우재;이대원
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.15-19
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    • 1999
  • For the induction of somatic embryogenic callus, the penduncle explants of Corydalis remota for peatinata were cultured on MS basal media supplemented with 2,4-D, kinetin and zeatin. The highest embryogenic callus formation was observed on the media containing 2.0 mg/L of 2,4-D and 2.0 mg/L of zeatin. The somatic embryogenesis on the media with 0.5 mg/L of cytokinin (zeatin or kinetin) were excellent under light condition, however somatic embryos abnormally developed into plantlets. Normal dicotyledonary plantlets were found on MS medium supplemented with 1.0 mg/L of zeatin. When MS medium with 2,4-D plus cytokinin and with BAP were used, the secondary somatic embryogenesis took place in root explants of the regenerants derived from in vitro somatic embryogenic callus.

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High frequency Somatic Embryogenesis and Plant Regeneration in Tissue Cultures of korean Cultivar Sweet Potatoes (체세포배발생에 의한 한국 고구마 품종의 고빈도 식물체 재분화)

  • 민성란;유장렬;노태홍;김칠현;주정일
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.157-160
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    • 1994
  • Culture conditions for high Sequency somatic embryogenesis and plant regeneration in tissue cultures of sweet potato of two Korean cultivars 'Puyojaerae' and 'Yulmi' are described. Shoot apical meristem explants (height 150 $\mu\textrm{m}$; base: 350 $\mu\textrm{m}$) were cultured on MS medium supplemented with 1 mg/L 2,4-D. After 6 weeks of culture, greater than 80% of the survived explants produced embryogenic calli. When transferred onto MS medium with 0.1 mg/L each of 2,4-D and kinetin, the calli gave rise to somatic embryos at frequencies of 71% ('Puyojaerae') and 63% ('Yulmi'), respectively: When somatic embryos at various developmental stages measured in length were transversely cut into two halves and cultured on MS medium with 1 mg/L 2,L-D, the upper halves produced secondary embryos more frequently than the lower ones, and halves of somatic embryos less than 1 mm in length had a higher competence for secondary embryo formation than longer ones of either cultivar. However 'Puyojaerae' somatic embryo halves showed a higher frequency of secondary embryo formation than 'Yulmi' ones on the whole. Upon transfer onto MS basal medium, most of the primary and secondary somatic embryos underwent development into plantlets. The plantlets were transplanted to potting soil and grown to maturity in a phytotron. The overall results suggest that the shoot apical meristem culture system for somatic embryo formation in sweet potato previously established by us (SABRAOJ 21: 93-101) may be applicable regardless of it genotypes.

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Plant Regeneration via Secondary Somatic Embryogenesis and Acclimatization in Panax ginseng (장뇌삼의 2차 체세포배 발생을 통한 식물체 유도 및 순화)

  • Lee, Su-Gwang;Kim, Ji-Hee;Kang, Ho-Duck
    • Journal of Korean Society of Forest Science
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    • v.97 no.1
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    • pp.127-133
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    • 2008
  • This study was conducted to establish the optimal condition for plant regeneration and acclimatization from somatic embryos of Panax ginseng. Cotyledon segments of Panax ginseng produced primary and secondary somatic embryos when cultured on MS and WPM media supplemented with 7% sucrose. To induce plantlet conversion, cotyledonary somatic embryos were cultured on WPM solid medium with $GA_3$ at various concentrations (1~30 mg/L) for 4 weeks. Plantlets were transferred to 1/2 WPM solid medium with $GA_3$ at various concentrations (0~5 mg/L) and 0.5% activated charcoal for shoot and root elongations. Elongated plantlets further developed into well-developed leaf and root system on 1/3 SH medium with 0.5% activated charcoal under ventilation condition for 5 months. The highest survival rate to soil was 75% when plantlets were regenerated on 1/3 SH medium without sucrose under ventilation condition.

Effect of Embryo Morphology on Plant Development in Secondary Somatic Embryogenesis of Alfalfa (알팔파 캘러스로부터 형성된 이차체세포배의 형태가 유식물 발달에 미치는 영향)

  • Won, S.H.;Lee, B.H.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.4
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    • pp.297-302
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    • 1999
  • Cotyledonary abnormalities were observed in secondary somatic embryos which were developed from calli cultured on MS medium with various concentrations of 2,4-D. In MS medium containing hormone-free or $0.1mg/{\ell}$ 2,4-D, the frequency of normal embryo formation with two cotyledons were above 57%. According to concentration of 2,4-D increment the frequency of normal embryos were decreased. In MS medium containing $4mg/{\ell}$ 2,4-D, the frequency of normal embryo formation was just 10%. The rate of germination was as follows; 37.7% of somatic embryos had one cotyledon, 85% two cotyledons, 38% three cotyledons, 35% four cotyledons, 25% five cotyledons and 29% trumpet-like cotyledons. About 80% of the embryos with two cotyledons were converted into normal plants, but one, three or four cotyledons were only 6.8~10%. The five or trumpet-like embryos were not developed into normal plants.

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Somatic embryogenesis and plant regeneration of Hovenia dulcis Thunb (헛개나무의 체세포배발생 및 식물체 재분화)

  • Eom, Seung-Hee;Shin, Dong-Yong;Lee, Hyeon-Yong;Kim, Myong-Jo;Kim, Jong-Dai;Choi, Won-Cheol;Heo, Kwon;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.1
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    • pp.41-45
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    • 2002
  • An efficient and reproducible procedure for the large scale propagation of Hovenia dulcis Thunb. is described. Shoot primodia emerging from the leaf surface was induced from MS medium supplemented with NAA. Stem cuttings were suitable explants for multiple shoot proliferation. They produced axillary shoots which branched repeatedly, yielding an average of 7 shoots per explants after 4 weeks in culture, when cultured on a woody plant medium (WPM) containing 0.1mg/l BA and 0.1mg/l NAA. Stem, leaf and root segments from axenic seedlings were used as explant source to induce somatic embryogenesis. A high frequency of somatic embryos were induced directly from leaf in MS medium with NAA, 2,4-D and in medium containing NAA, 2,4-D with BA. Somatic embryos were germinated in MS medium supplemented with 1mg/ l $GA_3$. Somatic embryos proliferated secondary somatic embryos rapidly after transfer to MS medium supplemented with 1mg/ l kinetin, 1mg/ l $GA_3$ and 2% dextrose.

Plant Regeneration via Somatic Embryogenesis and Organogenesis from Peucedanum japonicum $T_HUNB$ (갯기름나물의 체세포배 분화와 기관 분화를 통한 재분화)

  • Kim, Ok-Tae;Kim, Kwang-Soo;An, Jun-Chul;Hwang, Baik
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.21-24
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    • 2001
  • Peucedanum japonicum $T_{HUNB}$ used as a edible and medicinal plants was investigated for in uitro regeneration. Callus formation occurred on leaf and stem explant cultures and showed spontaneous embryogenic and organogenic capability on MS basal medium supplemented with 0.1~5 mg/L NAA and 0~10 mg/L BA in dark. The regeneration was highest on the condition supplemented with 2.5 mg/L NAA and 10 mg/L BA. Development of the somatic embryo progressed through the globular, heart-shaped, torpedo-shaped and cotyledonary stage, typical of zygotic embryos. When the first somatic embryos was cultured on the medium supplemented with 0.2 mg/L NAA, secondary somatic embryo were induced with higher frequency on the hypocotyl then on the cotyledon and root.t.

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Screening of Soybean Recombinant Inbred Lines for High Competence Somatic Embryogenesis (대두 재조합순계주에서 고빈도 체세포배발생능 계통 조사)

  • Choi, Pil-Son;Takao Komatsuda;Kim, Min-Hoon;Choi, Kyu-Myeong;Choi, Dong-Woog;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.135-138
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    • 2002
  • Cotyledonary explants from immature zygotic embryos of each 85 recombinant inbred lines (RILs) were cultured on medium containing MS salts, B5 vitamins, 40 mg$^{-1}$ 2,4-dichlorophenoxyacetic acid and 30 g$^{-1}$ sucrose. Frequency of somatic embryo formation on cotyledonary explants showed in thirty-six lines(<10%), in thirty-seven lines (11~49%), in nine lines (50~89%), and in three lines(>90%), respectively, The highest frequency (up to 90%) and number (6.36 per cotyledon) of somatic embryos were obtained from lines of KM1010, KM1032 and KM1064. Primary somatic embryos produced from three lines produced numerous secondary somatic embryos on the surfaces, which were subcultured for over one year. Upon transfer to maturation and conversion medium (Komatsuda, 1992), somatic embryos converted to plantlets at a frequency of approximately 25%.

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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