• Title/Summary/Keyword: Explants

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Effect of Cytokinins on in Vitro Growth of Grapes (Vitis spp.) (포도의 기내생장에 미치는 시토키닌의 영향)

  • Kim, Seung-Heui;Kim, Seon-Kyu
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.123-127
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    • 2002
  • Effect of cytokinins (BA, TDZ, zeatin, 2iP, and kinetin) applied either singly or in combination on in vitro growth of two grape cultivars ('Cabernet Sauvignon' and 'Campbell Early') was investigated as a serial work for mass production of grapevine nursery stocks. In single treatment, shoot growth of two cultivars was most favorable in control. Shoot proliferation was satisfactory with 10 $\mu$M BA regardless of cultivars and cytokinin combinations, followed by TDZ. Other treatments resulted in very poor or no branching. Total explants ready for subculture produced by 10 $\mu$M BA outnumbered those by other treatments. TDZ was also effective. TDZ significantly increased the fresh weight and callus formation while shoot growth was unsatisfactory. Shoot growth response of two cultivars in combined treatments was also most favorable in control as was in single treatments. When TDZ was combined with zeatin, 2iP, and kinetin which failed to induce branching, proliferous branching was induced though the shoot number was behind that of single treatments of BA and TDZ. TDZ was very effective for total number of explants and fresh weight, showing 10-fold increase.

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%.

Production of biomass and bioactive compounds from adventitious root cultures of Polygonum multiflorum using air-lift bioreactors (생물반응기를 이용한 적하수오 부정근의 바이오매스와 생리활성물질 대량생산)

  • Lee, Kyung-Ju;Park, Youngki;Kim, Ja-Young;Jeong, Taek-Kyu;Yun, Kyung-Seop;Paek, Kee-Yoeup;Park, So-Young
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.34-42
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    • 2015
  • This study was conducted to investigate the productivity of biomass and antioxidant compounds in Polygonum multiflorum by culturing explants in air-lift bioreactor containing Murashige and Skoog (MS) medium, by adding different concentrations of auxins [indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA)], sucrose, methyl jasmonate (MeJA), and salicylic acid (SA). Results of this study revealed that the explants culturing on the medium supplemented with $9.84{\mu}M$ IBA and 50 g/L sucrose were observed to have higher productivity of biomass and bioactive compound than other treatments used. Thus, we expect that these results will be helpful for large-scale production of biomass and antioxidant compounds from Polygonum multiflorum.

Characterization of in vitro Growth and Differentiation of an Albino Mutant of Nicotiana tobacum L. (Albino 담배 변이체의 기내 생장과 기내 분화의 특성)

  • ;;;;;;Yoshida Shigeo
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.197-203
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    • 1999
  • The albino plants of tobacco (Nicotiana tobacum L. cv. BY-4) were isolated from seed populations that were induced by heavy-ion ($^{14}N$) beam irradiation to proembryo and the in vitro growth and differentiation have been characterized. The in vitro cultured albino plants showed significant reduction of chlorophyll content and possessed larger number of stomata on both upper and lower epidermis than that of wild-type plants. Stem growth of the mutants remained dwarfed, however, the internode recovered its normal length after GA$_3$ treatment (10.0mg/L) on the MS medium containing sucrose under continuous light. When explants of leaf blades of albino plants were cultured, multiple shoots formed directly on MS medium containing 1.0mg/L of BAP or kinetin and a large number of calli were induced on the MS medium containing 1.0mg/L NAA or 1.0 mg/L 2,4-D. The albino calli regenerated multiple albino plantlets in the MS medium containing 0.1mg/L NAA + 1.0 mg/L BAP. No significant differences between the wild-type and albino plants were detected in the multiple shoot induction, callus formation from the explants and the plantlets regeneration from calli. In addition, albino plants have a similar organogenesis Pattern to that of the wild-type in the media with different combinations of NAA (0 to 5.0mg/L) and BAP (0 to 5.0mg/L) treatment. These results indicate that the albino mutant has the same normal regeneration ability as that of wild-type, although the mutant has lost functions in photosynthesis, such as pigmentation.

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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.

Antioxidative Activity and Flavonol Glycosides Analysis in Callus Derived from Leaf Tissue of Ginkgo biloba L. (은행(Ginkgo biloba L.)의 잎 유래 캘러스의 항산화능력 및 플로보놀 배당체 검정)

  • Kim, Jung-Suk;Park, Hye-Jeong;Park, Hyeon-Yong
    • Korean Journal of Plant Resources
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    • v.24 no.4
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    • pp.461-471
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    • 2011
  • This study was carried out to establish an in vitro culture method of callus having a high antioxidant activity from Ginkgo biloba L. Leaf explants were cultured on Murashige and Skoog's medium supplemented with various growth regulators. The explants were incubated in the dark or 3,000 lux cool-white light. Methanol extracts from incubated callus were evaluated for scavenging activity of the free radicals using DPPH. The best callus growth rate was achieved in MS medium combined with 10 ${\mu}M$ NAA and 5 ${\mu}M$ kinetin in the light condition. Total antioxidant activity of cell aggregates in suspension culture [MS medium supplemented with 10 ${\mu}M$ NAA in the light] was up to 80% of ascorbic acid. By means of HPLC analysis, quantification of the quercetin dehydrate and keamperol profiles from suspension callus was compared. Contents of quercetin dehydrate and keamperol from leaf extracts were 0.07 and 2.24 ${\mu}g/20{\mu}l$, and those from callus 0.56 and 0.18 ${\mu}g/20{\mu}l$, respectively.

Production of Phytol, an ACAT Inhibitor, from Callus Culture of Lettuce (Lactuca sativa L.) (상추 (Lactuca sativa L.) callus로부터 ACAT 억제 활성물질, phytol의 생산)

  • An, Kwang-Hee;Jang, Tae-O;Baek, Nam-In;Kim, Se-Young
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.63-68
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    • 2002
  • The possibility for mass production of phytol, inhibitory diterpene against ACAT (Acyl-CoA: Cholesterol acyltransferase) was investigated by using callus culture of lettuce. The callus were induced from lettuce cotyledon explants on MS medium containing 0.5 mg.L$^{-1}$ NAA after 4 week's culture. Adventitious roots were formed from the explants on MS medium containing 0.5 mg.L$^{-1}$ IBA or 1.0 mg.L$^{-1}$ NAA. Adventitious shoots and roots were emerged from the callus when the callus was transferred to MS medium containing auxin alone, or with cytokinin. The plant growth regulators and their concentrations for the organogenesis were 1.0 mg.L$^{-1}$ NAA, 0.1 mg.L$^{-1}$ BA, 0.5 mg.L$^{-1}$ NAA with 0.1 mg.L$^{-1}$ kinetin, or 0.5 g.L$^{-1}$ 2.4-D with 1.0 mg.L$^{-1}$ kinetin. Analyses of chlorophyll contents showed that chlorophyll contents were higher in morphogenic calli than in non-morphogenic calli. However, the chemical analyses of gas chromatography indicated that phytol contents were not proportionate to the chlorophyll contents of callus. The content of phytol was higher in callus than in lettuce cotyledon.ledon.

Factors Affecting the Agrobacterium Mediated Transformation of 'Gala' Apple (사과 갈라 품종의 Agrobacterium이용 형질전환에 영향하는 요인)

  • 송관정;성은수;황정환;제갈성;차지은;김정희;신용억
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.221-225
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    • 2001
  • Some factors of wounding methods, solidifying agents, origin of leaf explants, cone. of acetosyring-one, and MES affecting regeneration and transformation by Agrobacterium tumefaciens were investigated to establish an efficient transformation system of apple. Wounding by cutting the leaves merely showed the tendency of regeneration and transformation with higher efficiency compared with that of wounding by non-traumatic forcep when carrying out co-cultivation for three days after bacterial inoculation. While examining the solidifying agents of medium with the combination of agar (A)+Gelrit $e^{ }$ (G) in g. $L^{-1}$ , the higher concentration of Gelrit $e^{ }$ increased the efficiency of regeneration. However, there was no difference in the efficiency of transformation from the treatments of 2.5 G, 3.5 A+1.2 G, and 7.0 G. The origin of leaf explants showed no difference statistically in the efficiency of regeneration and transformation, but that from the shoots of proliferation medium showed the tendency with higher efficiency. The concentration of above 0.1 mM acetosyringon had an increase in the efficiency of regeneration and transformation and the concentration of 0.15 mM had the highest efficiency of transformation in the treatment of acetosyringone with different concentration. There was no effect of MES on regeneration and transformation.ion.

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Effects of different types and ages of explants and cytokinins on shoot regeneration in Brassica juncea L. (절편부위와 사이토키닌이 갓의 재분화에 미치는 영향)

  • Roh, Kyung Hee;Kwak, Bo-Kyung;Kim, Jong-Bum;Lee, Kyeong-Ryeol;Kim, Hyun Uk;Kim, Sun Hee;Kang, Han Chul
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.72-78
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    • 2013
  • To establish an efficient protocol for plant regeneration of Brassica juncea L. Czern, the effects of explant types, explant ages and cytokinins on shoot regeneration were examined in this study. Shoot regeneration was markedly affected by the explant types used in the following order: cotyledon with petiole> hypocotyl> leaf with petiole> cotyledon> leaf. Five-day-old seedlings of cotyledon with petiole explants showed the maximum shoot regeneration frequency. Of the six cytokinins-6-${\gamma}$-${\gamma}$-Dimethylallylamino-purine (2-ip), 6-${\gamma}$-${\gamma}$-Dimethylallylamino-purine riboside (2-ip riboside), 6-Benzyl amino-purine (BAP), Thidiazuron (TDZ), Zeatin, Zeatin riboside-TDZ ($8{\mu}M$) was found to be the best cytokinin for shoot regeneration with the highest shoot induction frequency (80%) from cotyledon with petiole after 4 weeks. All the regenerated plantlets were developed well and they produced morphologically normal flowers.

Efficient Transformation Method of Soybean Using Meristematic Tissues of Germinating Seeds (발아종자의 분열조직을 이용한 효율적인 콩 형질전환 방법)

  • Kim, Yul-Ho;Park, Hyang-Mi;Choi, Man-Soo;Sohn, Soo-In;Shin, Dong-Bum;Lee, Jang-Yong
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.278-285
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    • 2008
  • An efficient transformation method for soybean [Glycine max (L.) Merr.] using meristematic tissues of germinating seeds has been established. The embryonic axes were excised from germinating seeds of Korean soybean cultivar, Iksannamulkong and 0.5-2 cm long segment containing meristematic tissues were prepared by cutting hypocotyl region. The explants were inoculated with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector with the bar gene as a selectable marker gene and a ${\beta}-glucuronidase$ (GUSINT) reporter gene, and then co-cultured for 7 days on co-cultivation medium (CCM). The meristematic tissues were cultured on shoot induction medium (SIMP6) supplemented with 0.4 mg/l $N_6-benzylaminopurine$ (BAP) and 0.1 mg/l indolebutyric acid (IBA) in the presence of 6 mg/l L-phosphinotricin (PPT) for 2 weeks and the surviving explants were transferred to shoot elongation medium (SEMP6). Transformation was confirmed by Southern blot analysis and the transformation efficiencies ranged from 1.48 to 2.07%. The new modified transformation method was successfully implemented for obtaining several transgenic lines with SMV-CP gene. It is expected that this method could efficiently be used for the transformation of recalcitrant soybean cultivars.