Morphological Evaluation of Shoots Regenerated from Hygromycin-Resistant Rice Callus (cv IACuba-28)
An evaluation system based on the morphological characteristics of regenerated hygromycin-resistant rice callus shoots was established for correlating such characteristics with shoot viability on hygromycin. Embryogenic rice calli were transformed by Agrobacterium tumefaciens (EHA105/ pCAMBIA1300), containing the hygromycin-phosphotransferase gene as selection marker. After two weeks on selection medium, hygromycin-resistant calli were transferred to regeneration medium. Regenerated shoots were extracted every 5 days (over a 30-day period) and classified into three classes according to their morphological structure: class I: vigorous shoot having typical bipolar structure; class II: shoot having small root compared to apical length, or shoot without roots; class III: shoots having an abnormal appearance, such as malformed leaves or albinism. Individualised shoots were transferred to MS medium containing hygromycin for evaluating their resistance to antibiotics. A relationship was observed between regenerated shoots’ morphological characteristics and the percentage of shoots’ viability on hygromycin. Class I prevailed at early shoot extraction and was the most resistant to hygromycin. Drastic class I reduction was found with later shoot extraction, whilst classes II and III became increased. Likewise, shoot viability became radically reduced on MS medium containing hygromycin. This result might be applied for improving efficiency regarding obtaining transgenic rice plants, taking into account the best time for obtaining high percentages of hygromycin-resistant shoots having the best morphological characteristics.
| Main Authors: | , , , |
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| Format: | Digital revista |
| Language: | spa |
| Published: |
Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología
2007
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| Online Access: | https://revistas.unal.edu.co/index.php/biotecnologia/article/view/706 |
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| Summary: | An evaluation system based on the morphological characteristics of regenerated hygromycin-resistant rice callus shoots was established for correlating such characteristics with shoot viability on hygromycin. Embryogenic rice calli were transformed by Agrobacterium tumefaciens (EHA105/ pCAMBIA1300), containing the hygromycin-phosphotransferase gene as selection marker. After two weeks on selection medium, hygromycin-resistant calli were transferred to regeneration medium. Regenerated shoots were extracted every 5 days (over a 30-day period) and classified into three classes according to their morphological structure: class I: vigorous shoot having typical bipolar structure; class II: shoot having small root compared to apical length, or shoot without roots; class III: shoots having an abnormal appearance, such as malformed leaves or albinism. Individualised shoots were transferred to MS medium containing hygromycin for evaluating their resistance to antibiotics. A relationship was observed between regenerated shoots’ morphological characteristics and the percentage of shoots’ viability on hygromycin. Class I prevailed at early shoot extraction and was the most resistant to hygromycin. Drastic class I reduction was found with later shoot extraction, whilst classes II and III became increased. Likewise, shoot viability became radically reduced on MS medium containing hygromycin. This result might be applied for improving efficiency regarding obtaining transgenic rice plants, taking into account the best time for obtaining high percentages of hygromycin-resistant shoots having the best morphological characteristics. |
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