• Journal of Plant Biotechnology
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• v.40 no.3
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• pp.135-140
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• 2013

A less simple approach developed for generation of marker-free transgenic plants is to select transformants without the use of selective marker genes. Some results about development of marker-free transgenic plants were obtained using a non-selective approach in several crops such as rice, potato and tobacco. However, the study did not provide evidence on detailed characterization of introduced gene on genome, a critical step for confirming the stable integration and transmission of a foreign gene. In this study, we evaluated structure and integration sites of transgene (mCry1Ac) in the transgenic Bt rice plants which were made via conventional Agrobacterium-mediated transformation by non-selective method. Structure and integration sites of transgene in these transgenic plants had similar fashion as those recovered under selection.

• Journal of Plant Biotechnology
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• v.33 no.3
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• pp.185-194
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• 2006

Chloroplasts are believed to be descended from certain cyanobacteria, which were taken up by phagocytosis into a host cell and lived there in a symbiotic relationship. In contrast to the current static concept on the chloroplast genome, its dynamism has been recently demonstrated: the chloroplast genome is active in intramolecular homolgous recombination, producing subgenomic circles when it obtains homolgous sequences via genetic transformation. Chloroplast tranformation in higher plants provides many advantages over nuclear transformation that include higher expression levels of transgenes, polycistronic expression of transgenes, and maternal transmission of transgenes. Tobacco has been used as a model for chloroplast genetic transformation. However, it is recently possible to transform the chloroplasts of other major food and economic crops including rice, soybean, and cotton. Chloroplast-transformed crops will be able to replace bioreactors using microorganisms for production of value-added proteins in future.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.75-79
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• 2001

An experiment was carried out to introduce OsHSP17.9, a low molecular HSP gene isolated from rice plant to orchardgrass (Dactylis glomerata L.) using Agrobacterium. Mature seed-derived calli of orchardgrass were co-cultured with Agrobacterium tumefaciens EHA101 harboring the plasmid pIG-HSP17.9 for transformation. Calli selected by hygromycin were transferred to N$_{6}$ medium containing 1 mg/L NAA, 5 mg/L kinetin, 250 mg/L cefotaxime and 50 mg/L hygromycin and several hygromycin resistant plants were obtained. Stable incorporation of the introduced OsHSP17.9 to the genome of the hygromycin resistant plants was confirmed by PCR and Southern blot analysis. Transformation efficiency was variable between cultivars in which it was 16.5% in Potomac and 8.0% in Frontier. Constitutive expression of the transgene in the transformed orchardgrass tissues was identified by Northern blot analysis but transcript levels were different among individual plants.s.

• Journal of Plant Biotechnology
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• v.36 no.1
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• pp.23-29
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• 2009

We have previously isolated a CCCH type zinc-finger protein gene, OsZF2 (Oryza sativa Zinc Finger 2), from the cold-treated rice cDNA library. To investigate the potential role of OsZF2, transgenic rice lines over-expressing OsZF2 under the control of CaMV 35S promoter have been developed through Agrobacterium-mediated transformation. Elevated level of OsZF2 transcripts was confirmed by RNA gel blot analysis in transgenic rice. Under the 100 mM NaCl condition, the transgenic rice showed significantly enhanced growth rate in terms of shoot length and fresh weight, implicating that OsZF2 is likely to be involved in salt response of rice. In the field condition, however, the transgenic rice showed a dwarf phenotype and flowering time was delayed. Genome expression profiling analysis of transgenic plants using the 20K NSF rice oligonucleotide array revealed many up-regulated genes related to stress responses and signaling pathways such as chaperone protein dnaJ 72, salt stress-induced protein, PR protein, disease resistance proteins RPM1 and Cf2/Cf5 disease resistance protein, carbohydrate/ sugar transporter, OsWAK kinase, brassinosteroid LRR receptor kinase, and jasmonate O-methyltransferase. These data suggest that the CCCH type zinc-finger protein OsZF2 is a upstream transcriptional factor regulating growth and stress responsiveness of rice.

• Journal of Life Science
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• v.22 no.12
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• pp.1614-1620
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• 2012

Transgenic rice plants with increased resistance to rice brown planthopper (Nilaparvata lugens St${\aa}$l) were generated by particle bombardment-mediated transformation of plants with a gene encoding snowdrop lectin (Galanthus nivalis agglutinin; GNA) under control of the rice Rubisco small subunit (rbcS) promoter.. A large number of transgenic rice plants containing the GNA gene were generated. The integration, expression, and inheritance of this gene in the $R_1$ and $R_2$ generations were demonstrated by Southern and western blot analyses. The plants contained one to five copies of the transgene. The GNA protein comprised approximately 0.01-2.0% of total soluble protein in the $R_1$ and $R_2$ transgenic plants. Insect bioassays and feeding studies showed that the GNA protein expressed in the $R_2$ transgenic rice plants reduced the survival of brown planthoppers. The introduction of GNA into rice plants therefore can help to control insect pests.

• Journal of Life Science
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• v.24 no.6
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• pp.618-625
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• 2014

Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant $T_0$ plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the $T_1$ generation.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.540-546
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• 2010

This study was conducted to understand the role of soybean pathogenesis-related 10 (GmPR10) gene in salt tolerance and to develop salt-tolerant rice using GmPR10 cDNA. GmPR10 transgene was expressed constitutively in the shoot and root of the $T_1$ transgenic rice plants. Interestingly, however, the levels of the transgene expression were increased temporally up to over four- to five-fold in the shoot and root by 125 mM NaCl treatment, peaking at six hours after the treatment and decreasing thereafter. Electrolyte leakage of leaf cells under 125 mM NaCl treatment was lower in all the transgenic lines than in the control variety, Dongjin-byeo. Ability of seedlings to recover from 125 mM NaCl treatment for two weeks was higher in the transgenic plants than in the control plants. These results demonstrated that GmPR10 had function to increase cell integrity and promote growth under the saline stress imposed by NaCl. The transgenic line GmPR10-3 which showed highest ability to recover from the saline stress could be used as a potential source for salt tolerance in rice breeding programs.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.2
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• pp.59-63
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• 1998

Three different male-sterile cytoplasmic lines and their common maintainer 'Zhenshan 97B' and two elite restorer lines were used to study cytoplasmic effects on agronomic trait manifestation per se under different nitrogen supply levels. The result showed that cytoplasmic effects could be modified by nitrogen environments. The cytoplasmic effect on grain yield under 150 kg N/ha varied depending on crosses, while it was significantly negative in most crosses under both 60 and 330 kg N/ha. The correlation and path-coefficient analyses suggested that it was expected to improve cytoplasmic effects through reducing maximum tillers and increasing the percentage of productive tillers, leading to increased productive tillers and higher yield in hybrid rice by the aid of cultural practice and genetic transformation. This study also revealed that the same cytoplasm in different combinations had differential effect under the same nitrogen environment, indicating that cytoplasmic effect was produced by interaction of nuclear genes with cytoplasm rather than cytoplasm per se. These results indicated the usefulness of evaluating diverse cytoplasmic sources in various nuclear genotypes bred for hybrid rice breeding program. The finding also suggested that negative cytoplasmic effect could be effectively overcome by elite restorer lines through the interaction of nuclear genes with female cytoplasm.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.3
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• pp.264-272
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• 2011

It needs to develop high phosphate-available rice that is able to minimize environmental pollution caused by phosphate fertilizer. Then we have transformed 4 rice transporter genes, OsPT(Oryza sativa Phosphate Transporter)1, OsPT4, OsPT7 or OsPT8, to rice (Oryza sativa cv. Dongjin) via Agrobacterium-mediated transformation. We tested adaptation in the P-deficient condition of Dongjin (parental) and each transgenic line in the pot and the field conditions. Definite physiological changes have been observed in OsPTs transgenic lines including culm length, root formation and heading date. Phosphate uptake at harvesting stage was about three times higher in OsPT1-OX (overexpression) and OsPT4-OX than in Dongjin (wt) without P application. There are no variations in total phosphate-content of brown rice of OsPT1-OX in spite of high phosphate uptake. Practically the expression of OsPT1 has contributed to stabilize grain production without P fertilization in rice cultivation than Dongjin.

• Journal of Life Science
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• v.23 no.11
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• pp.1317-1324
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• 2013

High-molecular weight glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the wheat grain. We have produced marker-free transgenic rice plants containing a wheat Glu-1Bx7 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using the Agrobacterium-mediated co-transformation method. The Glu-1Bx7-own promoter was inserted into a binary vector for seed-specific expression of the Glu-1Bx7 gene. Two expression cassettes comprised of separate DNA fragments containing only Glu-1Bx7 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to the Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Glu-1Bx7 or HPTII was infected to rice calli at a 3:1 ratio of Glu-1Bx7 and HPTII, respectively. Then, among 216 hygromycin-resistant $T_0$ plants, we obtained 24 transgenic lines with both Glu-1Bx7 and HPTII genes inserted into the rice genome. We reconfirmed integration of the Glu-1Bx7 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the wheat Glu-1Bx7 were stably expressed in the rice $T_1$ seeds. Finally, the marker-free plants harboring only the Glu-1Bx7 gene were successfully screened at the $T_1$ generation.