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http://dx.doi.org/10.5010/JPB.2002.29.2.085

Investigation of Transformation Efficiency of Rice Using Agrobacterium tumefaciens and High Transformation of GPAT (glycerol-3-phosphate acyltransferase) Gene Relative to Chilling Tolerance  

Seo, Mi-Suk (College of Agricultural & Life Science, Cunchon National University)
Bae, Chang-Hyu (College of Agricultural & Life Science, Cunchon National University)
Choi, Dae-Ock (College of Natural Science, Sunchon National University)
Rhim, Seong-Lyul (College of Natural Science, Hallym University)
Seo, Suk-Chul (National Agricultural Science and Technology Institute, RDA)
Song, Pill-Soon (Kumho Life and Environmental Science Laboratory)
Lee, Hyo-Yeon (College of Agricultural & Life Science, Cunchon National University)
Publication Information
Journal of Plant Biotechnology / v.29, no.2, 2002 , pp. 85-92 More about this Journal
Abstract
This study has been focused on improving transformation efficiency of rice using Agrobacterium tumefaciens. We have demonstrated the effect of this system when the GPAT gene related to the cold-resistance was transferred by Agrobacterium tumefaciens in rice. Transformation conditions were modified using intron $\beta$-glucuronidase (GUS) expression as a reporter gene in the rice. In this study, mature seed-derived calli of rice (Oruza sativa L. cv. Dongjin) were pre-cultured for 3 days and then infected with Agrobacterium. When this infected calli were cultured in the dark for 10 days on co-cu]lure medium containing 50 mg/L of CaCl$_2$, 30 mg/L of acetosyringone, 2 mg/L of 2,4-D, 120 mg/L of betaine, high GUS expression was observed. In the present transformation system, the efficiency of transformation of GPAT gene was about 54%. Stable integration of GPAT gene into chromosomal DNA was proven by southern blot analysis of genomic DNA isolated from T$_{0}$ progenies. The progenies (T1 generation) derived from primary transformant of 5 lines were segregated with a 3 (resistant) : 1 (sensitive ratio) in medium containing hygromycin. This high frequency transformation system can be used as a useful tool in transformation of another monocotyledon.n.
Keywords
Agrobacterium; co-culture; GPAT gene; transformation efficiency;
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