Genetic transformation of sugarcane by microprojectile bombardment of young leaf rolls
| MSI05P4136 | |
| Mulleegadoo, K. D. Dookun-Saumtally, A. | |
| Genetic transformation of sugarcane by microprojectile bombardment of young leaf rolls | |
| periodical article | |
| 2005 | |
| Proc. int. Soc. Sug. Cane Technol. | |
| 25 (2): 579-584 | |
| En | |
| En Fr Sp | |
| The introduction of foreign genes in sugarcane by microprojectile bombardment has, so far, been mainly effected using embryogenic callus as target explants. Disadvantages of this method are that it can take several months to produce sufficient callus and regular subcultures are also required. An alternative method using bombardment of young leaf rolls was therefore attempted. About 3 mm-thick leaf rolls of variety M 292/70 were dissected from surface-sterilised sugarcane tops and pre-cultured for six to ten days on callus induction medium (CI3). They were bombarded with plasmid pAHC25 containing the gus and bar genes using the Bio-Rad PDS-1000/He Particle Delivery System. Six days after bombardment, the leaf rolls were transferred from CI3 medium onto regeneration medium supplemented with 1 mg/L bialaphos and they were subcultured onto fresh selective medium every two weeks. Out of 258 explants bombarded, several developed shoot primordia but well-established shoots were regenerated from six of them. Twenty-two plants were transferred to the glasshouse and 17 established ones were sprayed with 0.1 per cent Bastar four weeks later. Two plants were resistant to the herbicide and they were analyzed for the presence of the bar gene by PCR. A band corresponding to the expected amplifiable fragment of the gene was detected in them. Gene integration in these plants was confirmed by Southern blot hybridisation. These two plants also expressed the gus gene. Our study has demonstrated the potential of using primary explants of sugarcane variety M 292/70 as targets for microprojectile bombardment. This system will be further evaluated with other varieties. | |
| Sugarcane microprojectile bombardment genetic transformation genetic engineering embryogenic callus leaf rolls Bar gene | |
| Mauritius | |
| Sugarcane: Cane breeding and genetic improvement | |
| Genetic techniques and protocols | |
| 2005-02-22 | |
| en | |
| LIB | |
| CAT | |
| Biotech |