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http://dx.doi.org/10.5352/JLS.2014.24.6.618

Development of Marker-free Transgenic Rice Expressing the Wheat Storage Protein, Glu-1Dy10, for Increasing Quality Processing of Bread and Noodles  

Park, Soo-Kwon (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Shin, DongJin (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Hwang, Woon-Ha (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Hur, Yeon-Jae (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Kim, Tae-Heon (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Oh, Se-Yun (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Cho, Jun-Hyun (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Han, Sang-Ik (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Lee, Seung-Sik (Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI))
Nam, Min-Hee (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Park, Dong-Soo (Department of Functional Crop, National Institute of Crop Science, Rural Development Administration)
Publication Information
Journal of Life Science / v.24, no.6, 2014 , pp. 618-625 More about this Journal
Abstract
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.
Keywords
Co-transformation; high-molecular-weight glutenin subunit (HMW-GS) protein; marker-free transgenic rice; wheat;
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