• Title/Summary/Keyword: Plant Transformation

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Current status on plant molecular farming via chloroplast transformation (엽록체 형질전환 유래 분자 농업의 연구 동향)

  • Min, Sung-Ran;Jeong, Won-Joong;Kim, Suk-Weon;Lee, Jeong-Hee;Chung, Hwa-Jee;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.275-282
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    • 2010
  • Chloroplast transformation in higher plants offers many attractive advantages over nuclear transformation, including a high-level accumulation of foreign proteins, multi-gene expression in single transformation event via transgene stacking in operons and no position effect due to site-specific integration of transgenes by homologous recombination. Most importantly, chloroplast transgenic plants are eco-friendly because their transgenes are maternally inheritance in most crop plants. However, chloroplast transformation system has limited success in crops alike nuclear transformation. In the past two decades, great progress has been made to overcome the limitations of chloroplast transformation, thus expending chloroplast bioreactor to several important crops including soybean, carrot, lettuce, and oilseed. Therefore, it has become possible that chloroplast transformation of crops can be used not only for the improvement of agronomic traits, but also for the production of vaccines and high valuable therapeutic proteins in pharmaceutical industry.

The Effects of Physicochemical Factors and Cell Density on Nitrite Transformation in a Lipid-Rich Chlorella

  • Liang, Fang;Du, Kui;Wen, Xiaobin;Luo, Liming;Geng, Yahong;Li, Yeguang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2116-2124
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    • 2015
  • To understand the effects of physicochemical factors on nitrite transformation by microalgae, a lipid-rich Chlorella with high nitrite tolerance was cultured with 8 mmol/l sodium nitrite as sole nitrogen source under different conditions. The results showed that nitrite transformation was mainly dependent on the metabolic activities of algal cells rather than oxidation of nitrite by dissolved oxygen. Light intensity, temperature, pH, NaHCO3 concentrations, and initial cell densities had significant effects on the rate of nitrite transformation. Single-factor experiments revealed that the optimum conditions for nitrite transformation were light intensity: 300 μmol/m2/s; temperature: 30℃ pH: 7-8; NaHCO3 concentration: 2.0 g/l; and initial cell density: 0.15 g/l; and the highest nitrite transformation rate of 1.36 mmol/l/d was achieved. There was a positive correlation between nitrite transformation rate and the growth of Chlorella. The relationship between nitrite transformation rate (mg/l/d) and biomass productivity (g/l/d) could be described by the regression equation y = 61.3x (R2 = 0.9665), meaning that 61.3 mg N element was assimilated by 1.0 g dry biomass on average, which indicated that the nitrite transformation is a process of consuming nitrite as nitrogen source by Chlorella. The results demonstrated that the Chlorella suspension was able to assimilate nitrite efficiently, which implied the feasibility of using flue gas for mass production of Chlorella without preliminary removal of NOX.

Efficiency of transformation mediated by Agrobacterium tumefaciens using vacuum infiltration in rice (Oryza sativa L.)

  • Safitri, Fika Ayu;Ubaidillah, Mohammad;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.66-75
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    • 2016
  • Agrobacterium-mediated gene transfer has recently been developed to improve rice transformation. In this study, 3 different transformation methods were tested including soaking, co-cultivation, and vacuum infiltration. Agrobacterium tumefaciens GV3101 harboring the binary vector pGreen:: LeGSNOR was used in this experiment. This study aimed to identify the most appropriate method for transferring LeGSNOR into rice. Vacuum infiltration of the embryonic calli for 5 min in Ilpum resulted in high transformation efficiency based on confirmation by PCR, RT-PCR, and qRT-PCR analyses. In conclusion, we described the development of an efficient transformation protocol for the stable integration of foreign genes into rice; furthermore, the study results confirmed that PCR is suitable for efficient detection of the integrated gene. The vacuum infiltration system is a potentially useful tool for future studies focusing on transferring important genes into rice seed calli, and may help reduce time and effort.

An easy and efficient protocol in the production of pflp transgenic banana against Fusarium wilt

  • Yip, Mei-Kuen;Lee, Sin-Wan;Su, Kuei-Ching;Lin, Yi-Hsien;Chen, Tai-Yang;Feng, Teng-Yung
    • Plant Biotechnology Reports
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    • v.5 no.3
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    • pp.245-254
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    • 2011
  • This study describes an efficient protocol for Agrobacterium tumefaciens-mediated transformation of two subgroups of genotype AAA bananas (Musa acuminata cv. Pei Chiao and Musa acuminata cv. Gros Michel). Instead of using suspension cells, cauliflower-like bud clumps, also known as multiple bud clumps (MBC), were induced from sucker buds on MS medium containing $N^6$-Benzylaminopurine (BA), Thidiazuron (TDZ), and Paclobutrazol (PP333). Bud slices were co-cultivated with A. tumefaciens C58C1 or EHA105 that carry a plasmid containing Arabidopsis root-type ferredoxin gene (Atfd3) and a plant ferredoxin-like protein (pflp) gene, respectively. These two strains showed differences in transformation efficiency. The EHA105 strain was more sensitive in Pei Chiao, 51.3% bud slices were pflp-transformed, and 12.6% slices were Atfd3-transformed. Gros Michel was susceptible to C58C1 and the transformation efficiency is 4.4% for pflp and 13.1% for Atfd3. Additionally, gene integration of the putative pflp was confirmed by Southern blot. Resulting from the pathogen inoculation assay, we found that the pflp transgenic banana exhibited resistance to Fusarium oxysporum f. sp. cubense tropical race 4. This protocol is highly advantageous to banana cultivars that have difficulties in setting up suspension cultures for the purpose of quality improvement through genetic transformation. In addition, this protocol would save at least 6 months in obtaining explants for transformation and reduce labor for weekly subculture in embryogenic cell suspension culture systems.

Analysis of the Ripple Effect of Digital Transformation in the Shipbuilding & Offshore Plant Industry (조선해양플랜트산업의 디지털 전환에 따른 파급효과 분석)

  • Young-Gyu Lee;Woon-Seek Lee;Se-Hoon Park;Young-Seok Ock
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.46 no.spc
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    • pp.9-18
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    • 2023
  • In the global manufacturing industry, digital transformation is emerging as an important issue for companies to improve productivity and strengthen industrial competitiveness. The government and shipbuilding companies drive research and development to attain advanced technologies through digital transformation for shipbuilding & offshore plants, one of Korea's representative manufacturing industries. Therefore, the digital transformation of the shipbuilding & offshore plant industry was defined using an input-output analysis model, and the economic interrelationships of industries linked to the digital transformation of the shipbuilding & offshore plant industry were analyzed. According to the analysis, the shipbuilding & offshore plant industry ranks second among all 34 industries regarding the forward linkage effect. The backward linkage effect was ranked 27th, making it a mid-demand industry with a robust forward linkage effect. In addition, the production-induced effect was 0.455, the value-added-induced effect was 0.174, and the employment-induced effect was 1.779 people per billion won. The contribution of this study is that it will provide the basis for establishing policies necessary to strengthen the competitiveness of the shipbuilding & offshore plant industry. Also, it will help analyze the economic effects of digital transformation in other manufacturing industries.

Development of genetic transformation method of Korean soybean (국내콩 형질전환 기술개발)

  • Jeon, Eun-Hee;Chung, Young-Soo
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.344-351
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    • 2009
  • Current status of soybean transformation method in Korera was reviewed with recent publications. Most frequently used method for genetic transformation was Agrobacterium-mediated transformation on cotyledonary node which is most popular method used in foreign country. In addition to this, various methods such as sonicationmediated transformation, in planta transformation, and transformation on meristem tissue of germinating seed, have been tried in Korea, even though their efficiencies on repeatability and stability were relatively low. Based on the promising results developed recently by reviewer, several important considerations for successful soybean transformations were suggested. They are 1) proper genotype screening, 2) targeting transformation on exact point, 3) multiple shoot formation, 4) efficient selection pressure, 5) successful shoot elongation, 6) efficient root formation. These are the basic requirements for stable and highly efficient soybean transformation of Korean soybean.

Physical wounding-assisted Agrobacterium-mediated transformation of juvenile cotyledons of a biodiesel-producing plant, Jatropha curcas L.

  • Khemkladngoen, Naruemon;Cartagena, Joyce A.;Fukui, Kiichi
    • Plant Biotechnology Reports
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    • v.5 no.3
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    • pp.235-243
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    • 2011
  • The non-edible plant Jatropha curcas L. is one of the most promising feedstock for sustainable biodiesel production as it is not a source of edible vegetable oils, produces high amounts of oil (approx. 30-60% in dry seeds) and does not require high-cost maintenance. However, as with other undomesticated crops, the cultivation of J. curcas presents several drawbacks, such as low productivity and susceptibility to pests. Hence, varietal improvement by genetic engineering is essential if J. curcas is to become a viable alternative source of biodiesel. There is to date no well-established and efficient transformation system for J. curcas. In this study, we tested various physical wounding treatments, such as sonication and sand-vortexing, with the aim of developing an efficient Agrobacterium-mediated transformation for J. curcas. The highest stable transformation rate (53%) was achieved when explants were subjected to 1 min of sonication followed by 9 min of shaking in Agrobacterium suspension. The transformation frequency achieved using this protocol is the highest yet reported for J. curcas.

Molecular and Cytogenetic Analysis of Transgenic Plants of Rice(Oryza sativa L.) Produced by Agrobacterium-mediated Transformation

  • Cho, Joon-Hyeong;Kim, Yong-Wook
    • Plant Resources
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    • v.7 no.1
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    • pp.39-46
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    • 2004
  • To demonstrate the importance of transformation efficiency in independent event, molecular and cytogenetic analysis were conducted with genomic DNA and chromosome of transgenic plants produced by Agrobacterium tumefeciens LBA4404 (pSBM-PPGN: gusA and bar). Selection ratios of putative transgenic calli were similar in independent experiments, however, transformation efficiencies were critically influenced by the type of regeneration media. MSRK5SS-Pr regeneration mediun, which contains 5 mgL$^{-1}$ kinetin, 2% (w/v) sucrose in combination with 3% (w/v) sorbitol, and 500 mgL$^{-1}$ proline, was efficient to produce transgenic plant of rice from putative transgenic callus in the presence of L-phosphinotricin (PPT). With MSRK5SS-Pr medium, transformation efficincies of Nagdongbyeo were significantly enhanced from 3.7% to 6.3% in independent callus lines arid from 7.3% to 19.7% in plants produced, respectively. Stable integration and expression of bar gene were confirmed by basta herbicide assay, PCR amplification and Southern blotting of bar gene, and fluorescence in situ hybridization (FISH) analysis using pSBM-PPGN as a probe. In Southern blot analysis, diverse band patterns were observed in total 44 transgenic plants regenerated from 20 independent PPT resistant calli showing from one to five copies of T-DNA segments, however, the transformants obtained from one callus line showed the same copy numbers with the same fractionized band patterns.

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Lily Pollen Growth in vitro and Agrobacterium-mediated GUS Gene Transformation via Vacuum-Infiltration

  • Park, In-Hae;Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.151-154
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    • 2002
  • Conditions for lily pollen growth in vitro and transformation were optimized. Active pollen tube development was achieved effectively in a medium containing 7% sucrose with pH adjusted to 5.7 at the temperature of 27$^{\circ}C$ for about 16-24 hours. Pollen growth was little impaired by the presence of kanamycin at concentration up to 100 mg/L. Pollen rains near the beginning of germination stage were more reliable for Agrobacterium-mediated GUS DNA transformation via vacuum infiltration lasted for 20-40 minutes. GUS DNA integration and its expression in fully developed pollen tubes could be confirmed by Southern blot hybridization, RT-PCR and histochemical staining.

Effects of Sonication and Vacuum Infiltration on Agrobacterium-Mediated Transformation in Immature Embryos of Korean Wheat Genotypes

  • Moon Jung-Hun;Kang Moon-Suk;Heo Hwa-Young;Kwon Young-Up;Lee Sang-Kyu;Lee Kyung-Hee;Lee Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.415-418
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    • 2004
  • The effects of sonication and vacuum infiltration on transformation efficiency was investigated by using immature embryos of Korean wheat as explants. Two Agrobacterium tumefaciens strains, KYRT1 and EHA105, carrying pCAMBIA 1305.1 were used. Transformation efficiency was demonstrated by the detection of $\beta-glucu-ronidase$ (GUS) activity. GUS expression showed clear difference among Korean wheat cultivars. Geurumil showed higher GUS expression efficiency $79.1\%$ compared with other cultivars. The effects of the duration of vacuum infiltration and sonication treatment showed a tendency high GUS expression efficiency by their combination. In comparison with other Agrobacterium strains, KYRT1 showed high efficiency in most Korean cultivars.