Genetic transformation of Curcuma alismatifolia Gagnep. using retarded shoots
| Autor: | S. Mahadtanapuk, M. Sanguansermsri, Somboon Anuntalabhochai, Nopmanee Topoonyanont, Takashi Handa |
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| Rok vydání: | 2006 |
| Předmět: | |
| Zdroj: | Plant Biotechnology. 23:233-237 |
| ISSN: | 1347-6114 1342-4580 |
| Popis: | A protocol for regeneration and genetic transformation was established for Curcuma alismatifolia Gagnep. 'Chiang Mai Pink' using retarded shoots as explants. In vitro retarded shoots were cut into 0.5� 0.5� 0.5 cm blocks and co- cultivated with Agrobacterium tumefaciens strain AGLO harboring the binary vector, pBI121 or pBI121-Ca-ACS1. The explants were incubated in the bacteria suspension for 30 min. The explants and bacteria were cultured on MS medium for 2 days in darkness at 25°C for co-cultivation. Then, the explants were transferred onto MS medium containing 0.1 mg l � 1 IAA, 4 mg l � 1 IMA, 0.5 mg l � 1 TDZ, 50 mg l � 1 kanamycin and 500 mg l � 1 vancomycin. The explants were subcultured every 2 weeks. After 4 weeks in culture, the explants with small shoot buds were transferred onto MS medium containing 50 mg l � 1 kanamycin. Within 4 weeks, the shoots were separated and subcultured every 2 weeks on MS medium containing 0.1 mg l � 1 IAA and 50 mg l � 1 kanamycin. PCR analysis, histochemical GUS assay and Southern blotting of the regenerated plants confirmed transformation events. We obtained transformed plants within 3 months after co-cultivation with the bacteria and the transformation frequency exceeded 14%, which is suitable for practical use. |
| Databáze: | OpenAIRE |
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