• Title/Summary/Keyword: plant transformation

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Single Somatic Embryogenesis from Transformant with Proteinase II Gene in Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Kim, Se-Young;Rho, Yeong-Deok;Kim, Moo-Sung
    • Plant Resources
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    • v.6 no.3
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    • pp.205-210
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    • 2003
  • Ginseng(Panax ginseng C.A. Meyer) is a perennial herbaceous plant which grows very slowly. It takes about 3 to 4 years from seeding to collecting the ripe seeds and the ginseng propagation is very difficult. and so, it is very difficult to breed ginseng plant. Ginseng tissue culture was started from at 1960, and ginseng commercial product by in vitro callus culture was saled, however upto now, regenerants were not planted to soil normally. Recently, plant genetic engineering to produce transgenic plants by introducing useful genes has been advanced greatly. In a present paper, transformation of ginseng plants was achieved by co-cultivation with Agrobacterium harboring the binary vector coding Proteinase-II gene, which confer resistant or tolerant to insect pests, The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 35S promoter. The NPT II gene and introduced genes of the transgenic ginseng plants were successfully identified by the PCR. Especially the transgenic ginseng plants were regenerated using new techniques such as repetitive single somatic embryogenesis.

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Usage of Tobacco Plants for Various Purposes (담배 속 식물의 다양한 활용방안 모색)

  • Um, Yu-Rry;Lee, Moon-Soon;Lee, Yi;Seok, Yeong-Seon
    • Journal of the Korean Society of Tobacco Science
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    • v.33 no.1
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    • pp.8-15
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    • 2011
  • Genus Nicotiana has 76 species including N. tabacum. These plants are used not only as a material for cigarette manufacturing but also as ornamental plant, medicinal plant, poisonous substance plant, and bug repellent plant. N. tabacum is used as a main material for cigarette manufacturing with N. rustica. N. sylvestris and N. alata is used as ornamental plants because of their beautiful flowers and N. rustica is used for bug repellent or pesticide because of its high concentration of nicotine. N. glauca, a tree tobacco, is used for bio-fuel production. N. tabacum is used as a popular model plant system for degeneration, regeneration, and transformation. N. benthamiana is also used as a model system for foreign gene expression by agroinfiltration. The transformation ability of tobacco plant is a good target for molecular farming. Hepatitis B virus envelop protein, E. coli heat-labile enterotoxin, diabetes autoantigen, and cholera toxin B subunit were produced using tobacco plants. Secondary metabolites of tobacco include nicotine, anabasine, nornicotine, anatabine, cembranoid, solanesol, linoleic acid, rutin, lignin and sistosterol, and they are used for various medicine productions which cannot be produced by organic synthesis for their complicated structures. In conclusion, we have to understand the applicability of tobacco plant in detail and study to enlarge the usage of the plants.

Production of Transgenic Petunia hybrida cv. Rosanpion Using Agrobacterium-mediated Transformation

  • Ko, Jeong-Ae;Kim, Young-Sook;Kim, Myung-Jun;Kim, Hyun-Soon
    • Plant Resources
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    • v.4 no.1
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    • pp.36-40
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    • 2001
  • Transgenic Petunia hybrida cv. Rosanpion was produced by Agrobactepium tumefaciens LBA4404 harboring a binary vector pBI 121 containing $\beta$-glucuronidase (gus) and neomycin phosphotransferase (nptII). For genetic transformation, leaf discs were precultured on MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA (MNB) for 2 days and cocultured for 15 mins with A. tumefaciens. For selection of transformant, leaf discs were transferred to fresh MNB containing 50 mg/L kanamycin and 500 mg/L cefotaxime. Eighteen plants were regenerated and four were confirmed by PCR for detection of gus and nptII gene integrated into the nuclear genome of petunia ‘Rosanpion’. Using this transformation system, we expect that transgenic petunia ‘Rosanpion’ incorporating a useful gene can be produced.

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Mitochondrial Porin Isoform AtVDAC1 Regulates the Competence of Arabidopsis thaliana to Agrobacterium-Mediated Genetic Transformation

  • Kwon, Tackmin
    • Molecules and Cells
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    • v.39 no.9
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    • pp.705-713
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    • 2016
  • The efficiency of Agrobacterium-mediated transformation in plants depends on the virulence of Agrobacterium strains, the plant tissue culture conditions, and the susceptibility of host plants. Understanding the molecular interactions between Agrobacterium and host plant cells is crucial when manipulating the susceptibility of recalcitrant crop plants and protecting orchard trees from crown gall disease. It was discovered that Arabidopsis voltage-dependent anion channel 1 (atvdac1) mutant has drastic effects on Agrobacterium-mediated tumorigenesis and growth developmental phenotypes, and that these effects are dependent on a Ws-0 genetic background. Genetic complementation of Arabidopsis vdac1 mutants and yeast porin1-deficient strain with members of the AtVDAC gene family revealed that AtVDAC1 is required for Agrobacterium-mediated transformation, and there is weak functional redundancy between AtVDAC1 and AtVDAC3, which is independent of porin activity. Furthermore, atvdac1 mutants were deficient in transient and stable transformation by Agrobacterium, suggesting that AtVDAC1 is involved in the early stages of Agrobacterium infection prior to transferred-DNA (T-DNA) integration. Transgenic plants overexpressing AtVDAC1 not only complemented the phenotypes of the atvdac1 mutant, but also showed high efficiency of transient T-DNA gene expression; however, the efficiency of stable transformation was not affected. Moreover, the effect of phytohormone treatment on competence to Agrobacterium was compromised in atvdac1 mutants. These data indicate that AtVDAC1 regulates the competence of Arabidopsis to Agrobacterium infection.

Establishment of Genetic Transformation System and Introduction of MADS Box Gene in Hot Pepper (Capsicum annuum L.)

  • Lim, Hak-Tae;Zhao, Mei-Ai;Lian, Yu-Ji;Lee, Ji-Young;Eung-Jun park;Chun, Ik-Jo;Yu, Jae-Woong;Kim, Byung-Dong
    • Journal of Plant Biotechnology
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    • v.3 no.2
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    • pp.89-94
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    • 2001
  • In vitro plant regeneration of inbred breeding line of hot pepper (Capsicum annuum L.) was established using leaf and petiole segments as explants. About 28 days old plants were excised and cultured on MS medium supplemented with TDZ and NAA or in combination with Zeatin. In all of the media compositions tested, combination of TDZ 0.5 mg/L, Zeatin 0.5 mg/L, and NAA 0.1 mg/L was found to be the best medium for shoot bud initiation. Young petiole was the most appropriate explant type for the plant regeneration as well as genetic transformation in hot pepper. In this study, HpMADS1 gene isolated from hot pepper was introduced using Agrobacterium-mediated transformation system. Based on the analysis of Southern blot and RT-PCR, HpMADS1 gene was integrated in the hot pepper genome. It has been known that floral organ development is controlled by a group of regulatory factors containing the MADS domain. Morphological characteristics in these transgenic plants, especially flowering habit, however, were not significantly altered, indicating this MADS gene, HpMADS1 may be non-functional in this case.

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Effects of Antibiotics on Suppression of Agrobacterium tumefaciens and Plant Regeneration from Wheat Embryo

  • Han, Si-Nae;Oh, Poo-Reum;Kim, Hong-Sig;Heo, Hwa-Young;Moon, Jun-Cheol;Lee, Sang-Kyu;Kim, Kyung-Hee;Seo, Yong-Weon;Lee, Byung-Moo
    • Journal of Crop Science and Biotechnology
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    • v.10 no.2
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    • pp.92-97
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    • 2007
  • Antibiotics used for suppressing Agrobacterium in plant transformation procedure might have negligible effects on plant tissues and regeneration. The effects of antibiotics on growth suppression of Agrobacterium and plant regeneration were investigated for enhancing Agrobacterium-mediated transformation using wheat mature embryos. Antibiotics tested, except carbenicillin, were able to suppress that embryos were coated with a layer of Agrobacterium cells in callus induction medium. Agrobacterium growth was suppressed minimally at 50 mg/l of timentin, while cefotaxime and clavamox were completely suppressed at relative high concentration of 250 mg/l. In the treatment of carbenicillin, initiation of growth suppression of Agrobacterium occurred at 750 mg/l of concentration because Agrobacterium KYRT1 contains the carbenicillin resistant gene. In Agrobacterium inoculation, effects of antibiotics were significantly different on the rate of callus induction and shoot formation. Almost embryos were induced calli at 50 mg/l of timentin whereas callus induction rate was achieved above 90% at 100 mg/l and 250 mg/l of cefotaxime and clavamox, respectively. Shoot formation rate was higher in the treatment of timentin than that of cefotaxime and clavamox at 500 mg/l of concentration, respectively. Timentin can be used as a good antibiotics in Agrobacterium-mediated wheat transformation.

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A novel technique for recombinant protein expression in duckweed (Spirodela polyrhiza) turions

  • Chanroj, Salil;Jaiprasert, Aornpilin;Issaro, Nipatha
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.156-164
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    • 2021
  • Spirodela polyrhiza, from the Lemnaceae family, are small aquatic plants that offer an alternative plant-based system for the expression of recombinant proteins. However, no turion transformation protocol has been established in this species. In this study, we exploited a pB7YWG2 vector harboring the eYFP gene that encodes enhanced yellow fluorescent protein (eYFP), which has been extensively used as a reporter and marker to visualize recombinant protein localization in plants. We adopted Agrobacterium tumefaciens-mediated turion transformation via vacuum infiltration to deliver the eYFP gene to turions, special vegetative forms produced by duckweeds to endure harsh conditions. Transgenic turions regenerated several duckweed fronds that exhibited yellow fluorescent emissions under a fluorescence microscope. Western blotting verified the expression of the eYFP protein. To the best of our knowledge, this is the first report of an efficient protocol for generating transgenic S. polyrhiza expressing eYFP via Agrobacterium tumefaciens-mediated turion transformation. The ability of turions to withstand harsh conditions increases the portability and versatility of transgenic duckweeds, favoring their use in the further development of therapeutic compounds in plants.