• Title/Summary/Keyword: transformed plants

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Molecular Control of Gene Co-suppression in Transgenic Soybean via Particle Bombardment

  • El-Shemy, Hany A.;Khalafalla, Mutasim M.;Fujita, Kounosuke;Ishimoto, Masao
    • BMB Reports
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    • v.39 no.1
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    • pp.61-67
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    • 2006
  • Molecular co-suppression phenomena are important to consider in transgene experiments. Embryogenic cells were obtained from immature cotyledons and engineered with two different gene constructs (pHV and pHVS) through particle bombardment. Both constructs contain a gene conferring resistance to hygromycin (hpt) as a selective marker and a modified glycinin (11S globulin) gene (V3-1) as a target. sGFP(S65T) as a reporter gene was, however, inserted into the flanking region of the V3-1 gene (pHVS). Fluorescence microscopic screening after the selection of hygromycin, identified clearly the expression of sGFP(S65T) in the transformed soybean embryos bombarded with the pHVS construct. Stable integration of the transgenes was confirmed by polymerase chain reaction (PCR) and Southern blot analysis. Seeds of transgenic plants obtained from the pHV construct frequently lacked an accumulation of endogenous glycinin, which is encoded by homologous genes to the target gene V3-1. Most of the transgenic plants expressing sGFP(S65T) showed highly accumulation of glycinin. The expression of sGFP(S65T) and V3-1 inherits into the next generations. sGFP(S65T) as a reporter gene may be useful to increase the transformation efficiency of transgenic soybean with avoiding gene co-suppression.

Production of transgenic Alstroemeria plants containing virus resistance genes via particle bombardment

  • Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.164-171
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    • 2020
  • Transgenic Alstroemeria plants resistant to Alstroemeria mosaic virus (AlMV) were generated through RNA-mediated resistance. To this end, the friable embryogenic callus (FEC) of Alstroemeria was induced from the leaf axil tissue and transformed with a DNA fragment containing the coat protein gene and 3'-nontranslated region of AlMV through an improved particle bombardment system. The bar gene was used as a selection marker. More than 300 independent transgenic FEC lines were obtained. Among these, 155 lines resistant to phosphinothricin (PPT) were selected under low stringent conditions. After increasing the stringency of PPT selection, 44 transgenic lines remained, and 710 somatic embryos from these lines germinated and developed into shoots. These transgenic shoots were then transferred to the greenhouse and challenged with AlMV. In total, 25 of the 44 lines showed some degree of resistance. PCR analysis confirmed the presence of the viral sequence. Virus resistance was observed at various levels. Establishment of an efficient transformation system for Alstroemeria will allow inserting transgenes into this plant to confer resistance to viral and fungal pathogens. Accordingly, this is the first report on the production of a transgenic virus-resistant Alstroemeria and lays the foundation for alternative management of viral diseases in this plant.

Agrobacterium tumefaciens Mediated Genetic Transformation of Pigeonpea [Cajanus cajan (L.) Millsp.]

  • Kumar, S.Manoj;Syamala, D.;Sharma, Kiran K.;Devi, Prathibha
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.69-75
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    • 2004
  • Optimal protocol for efficient genetic transformation has been defined to aid future strategies of genetic engineering in pigeon pea with agronomically important genes. Transgenic pigeonpea plants were successfully produced through Agrobacterium tumefaciens-mediated genetic transformation method using cotyledonary node explants by employing defined culture media. The explants were co-cultivated with A. tumefaciens strain C-58 harboring the binary plasmid, pCAMBIA-1301 [con-ferring $\beta$-glucuronidase(GUS) activity and resistance to hygromycin] and cultured on selection medium (regeneration medium supplemented with hygromycin) to select putatively transformed shoots. The shoots were then rooted on root induction medium and transferred to pots containing sand and soil mixture in the ratio of 1:1. About 22 putative TO transgenic plants have been produced. Stable expression and integration of the transgenes in the putative transgenics were confirmed by GUS assay, PCR and Southern blot hybridization with a transformation efficiency of over 45%. Stable integration and expression of the marker gene has been confirmed in the TO and T1 transgenics through PCR, and Southern hybridization.

Modification of amylose content of sweetpotato starch by RNAi technique

  • Shimada, Takiko;Otani, Motoyasu;Hamada, Tatsurou;Kim, Sun-Hyung
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.351-355
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    • 2005
  • In the storage roots of sweetpotato (Ipomoea batatas (L.) Lam. cv. Kokei 14), 10 to 20% of starch is essentially unbranched linear amylose and the other major component is branched amylopectin. Amylose is produced by the enzyme GBSSI (granule bound starch synthase I), whereas amylopectin is produced by a concerted action of soluble starch synthase and starch branching enzymes (SBEI and SBEII). We constructed double-stranded RNA (dsRNA) interference vectors of GBSSI and IbSBEII and introduced them into sweetpotato genome via Agrobacterium-mediated gene transformation. The endogenous GBSSI expression was inhibited by dsRNA of GBSSI in 73 % of transgenic plants giving rise to the storage tubers containing amylopectin but not amylose. On the other hand, all sweetpotato plants transformed with dsRNA of IbSBEII contained a larger amount of amylose than the non-transgenic control (up to 25% compared to 10% in the controls). The RNA interference (RNAi) is effectively inhibited the gene expression in thestarch metabolic pathway and modified the characteristics of starch in sweetpotato.

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Development of transgenic disease-resistant root stock for the growth of watermelon

  • Cho, Song-Mi;Chung, Soo-Jin;Moon, Sun-Jin;Kim, Kwang-Sang;Kim, Young-Cheol;Cho, Baik-Ho
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2004.10a
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    • pp.62-65
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    • 2004
  • To protect the watermelon against soil-borne pathogens, we are currently producing disease-resistant transgenic root stock for the growth of watermelon, A defensin gene (J1-1) from Capsicum annum, a ACC deaminase gene from Pseudomonas syringae, a galactinol synthase (CsGolS) gene from Cucumis sativus, and a WRKY (CvWRKY2) gene from Citullus vulgaris were used as transgenes for disease resistance. The gene were transformed into a inbred line (6-2-2) of watermelon, Kong-dae watermelon and a inbred line (GO702S) of gourd, respectively, by Agrobacterium-mediated transformation. Putative transgenic plants were selected in medium containing 100mg/L kanamycin, and then integration of the genes into the genomic DNA were demonstrated by PCR analysis. Successful integration of the gene in regenerated plants was also confirmed by PCR (Figf 1), genomic Southern blot (Fig 2), RT-PCR (Fig 3), and Northern blot analysis(Fig 4). Several T1 lines having different transgene were produced, and disease resistance of the T1 lines are under estimation.

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Expression of Dengue virus EIII domain-coding gene in maize as an edible vaccine candidate

  • Kim, Hyun A;Kwon, Suk Yoon;Yang, Moon Sik;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.50-55
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    • 2014
  • Plant-based vaccines possess some advantages over other types of vaccine biotechnology such as safety, low cost of mass vaccination programs, and wider use of vaccines for medicine. This study was undertaken to develop the transgenic maize as edible vaccine candidates for humans. The immature embryos of HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vectors (V662 or V663). The vectors carrying nptII gene as selection marker and scEDIII (V662) or wCTB-scEDIII (V663) target gene, which code EIII proteins inhibite viral adsorption by cells. In total, 721 maize immature embryos were transformed and twenty-two putative transgenic plants were regenerated after 12 weeks selection regime. Of them, two- and six-plants were proved to be integrated with scEDIII and wCTB-scEDIII genes, respectively, by Southern blot analysis. However, only one plant (V662-29-3864) can express the gene of interest confirmed by Northern blot analysis. These results demonstrated that this plant could be used as a candidated source of the vaccine production.

Thinking multiculturality in the age of hybrid threats: Converging cyber and physical security in Akkuyu nuclear power plant

  • Bicakci, A. Salih;Evren, Ayhan Gucuyener
    • Nuclear Engineering and Technology
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    • v.54 no.7
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    • pp.2467-2474
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    • 2022
  • Nuclear Power Plants (NPPs) are the most protected facilities among all critical infrastructures (CIs). In addition to physical security, cyber security becomes a significant concern for NPPs since swift digitalization and overreliance on computer-based systems in the facility operations transformed NPPs into targets for cyber/physical attacks. Despite technical competencies, humans are still the central component of a resilient NPP to develop an effective nuclear security culture. Turkey is one of the newcomers in the nuclear energy industry, and Turkish Akkuyu NPP has a unique model owned by an international consortium. Since Turkey has limited experience in nuclear energy industry, specific multinational and multicultural characteristics of Turkish Akkuyu NPP also requires further research in terms of the Facility's prospective nuclear security. Yet, the link between "national cultures" and "nuclear security" is underestimated in nuclear security studies. By relying on Hofstede's national culture framework, our research aims to address this gap and explore possible implications of cross-national cultural differences on nuclear security. To cope with security challenges in the age of hybrid threats, we propose a security management model which addresses the need for cyber-physical security integration to cultivate a robust nuclear security culture in a multicultural working environment.

Global trends in regulatory frameworks for animal genome editing in agriculture

  • Dajeong Lim;Inchul Choi
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.4
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    • pp.247-253
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    • 2023
  • Revolutionary advancements, such as the reduction in DNA sequencing costs and genome editing, have transformed biotechnology, fostering progress in manipulating biomolecules, engineering cells, and computational biology. Agriculture and food production have significantly benefited from tools like high-throughput microarrays, accelerating the selection of desired traits. Genetic engineering, especially utilizing genome editing, facilitates precise alterations in plants and animals, harnessing microbiomes and fostering lab-grown meat production to alleviate environmental pressures. The emergence of new biotechnologies, notably genome editing, underscores the necessity for regulatory frameworks governing LM (living modified) organisms. Global regulations overseeing genetically engineered or genome-edited (GE) organisms, encompassing animals, exhibit considerable diversity. Nonetheless, prevailing international regulatory trends typically exclude genomeedited plants and animals, employing novel biotechnological techniques, from GMO/ LMO classification if they lack foreign genes and originate through natural mutations or traditional breeding programs. This comprehensive review scrutinizes ongoing risk and safety assessment cases, such as genome-edited beef cattle and fish in the USA and Japan. Furthermore, it investigates the limitations of existing regulations related to genome editing in Korea and evaluates newly proposed legislation, offering insights into the future trajectory of regulatory frameworks.

Selection of transgenic sweetpotato plants expressing 2-Cys peroxiredoxin with enhanced tolerance to oxidative stress (Peroxiredoxin 유전자 발현 산화스트레스 내성 형질전환 고구마의 선발)

  • Kim, Myoung-Duck;Yang, Kyoung-Sil;Kwon, Suk-Yoon;Lee, Sang-Yeol;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.75-80
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    • 2009
  • In order to develop transgenic sweetpotato plants [Ipomoea batatas (L.) Lam. cv. Yulmi] with enhanced tolerance to oxidative stress, we constructed transformation vectors expressing 2-Cys peroxiredoxin (Prx) gene under the control of the stress-inducible SWPA2 or enhanced 35S promoter (named as SP or EP). Transgenic sweetpotato plants were attempted to generate from embryogenic calli using an Agrobacterium-mediated transformation system. Embryogenic calli gave rise to somatic embryos and then converted into plantlets on MS medium containing 100 mg/L kanamycin. Transgenic plants were regenerated in the same medium. Southern blot analysis confirmed that the Prx gene was inserted into the genome of the plants. To further study we selected the transgenic plant lines with enhanced tolerance against methyl viologen (MV). When sweetpotato leaf discs were subjected to methyl MV at $20{\mu}M$, transgenic plants showed about 40% higher tolerance than non-transgenic or empty vector-transformed plants.

Tissue Culture of Stone Fruit Plants Basis for Their Genetic Engineering

  • Csanyi, Marta;Wittner, Anita;Nagy, Agenes;Balla, Ildiko;Vertessy, Judit;Palkovics, Laszlo;Balazs, Eevin
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.91-95
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    • 1999
  • Genetic engineering of stone fruit species like apricot, plum, peach and cherry are hampered by the inefficient and low-level regeneration processes in tissue culture. The first transgenic stone fruit species have emerged from transformed hypocotyls. These great achievements were applauded by the scientific community contrary the fact that hypocotyl derived transgenic plants have no real brooding value. Tissue culture of different organs of valuable cultivars are recorded with an extremely low-level of regeneration in the literature. To improve the tissue culture basis of stone fruit plants an extensive tissue culture programme were launched and dozens of different media were compared including a series of hormone concentration in the tissue culture systems. Our continuous efforts were crowned by a very efficient method for achieving up to 30-40% regenerable petioles. Usually on a single petiole several well-separated meristems were induced. After 3-4 weeks of cultivation shoots were developed. The basic media $K_2$ were supplemented with 10g/L saccharose, 10g/L glucose and 10g/L maltose. The following plant hormones were used BAP 1mg/L, TDZ 1mg/L, 2-iP 1mg/L and IAA 0,1 mg/L concentrations. The Petri dishes were kept for 3 weeks in dark at a temperature 22$^{\circ}C$ for 8 hours and 22-24$^{\circ}C$ for 16 hours. The Petri dishes were sealed with Parafilm. The regeneration of the petioles were genotype independent and we were able to regenerate different plum cultivars with almost the same efficiency.

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