Successful genetic transformation of Chinese cabbage using phosphomannose isomerase as a selection marker |
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Authors: | Byung-Whan Min Yi-Nam Cho Min-Jung Song Tae-Kyung Noh Bong-Kyu Kim Won-Ki Chae Young-Soo Park Yang-Do Choi Chee-Hark Harn |
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Institution: | (1) Department of Plant Resources, Sangju National University, Sangju, Gyeongbuk, Korea;(2) Biotechnology Institute, Nong Woo Bio Co. Ltd., Yeoju, Gyeonggi, Korea;(3) School of Agricultural Biotechnology, Seoul National University, Seoul, Korea |
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Abstract: | A mannose selection system was adapted for use in the Agrobacterium-mediated transformation of Chinese cabbage. This system makes use of the pmi gene that encodes phosphomannose isomerase, which converts mannose-6-phosphate to fructose-6-phosphate. Hypocotyl explants
from 4–5-day-old seedlings of Chinese cabbage inbred lines were pre-cultured for 2–3 days and then infected with Agrobacterium. Two genes (l-guluno-γ-lactone oxidase, GLOase, and jasmonic methyl transferase, JMT) were transformed into Chinese cabbage using the transformation procedure developed in this study. We found that supplementing
the media with 7 g l−1 mannose and 2% sucrose provides the necessary conditions for the selection of transformed plants from nontransformed plants.
The transformation rates were 1.4% for GLOase and 3.0% for JMT, respectively. The Southern blot analysis revealed that several independent transformants (T
0) were obtained from each transgene. Three different inbred lines were transformed, and most of the T
1 plants had normal phenotypes. The transformation method presented here for Chinese cabbage using mannose selection is efficient
and reproducible, and it can be useful to introduce a desirable gene(s) into commercially useful inbred lines of Chinese cabbage. |
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Keywords: | Chinese cabbage Mannose Phosphomannose isomerase Transformation Sucrose |
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