Citation
Masani M. Y. A., . and Rasid O. A., . and Bahariah B., . and Rahmah A. R. S., . and Ghulam Kadir Ahmad Parveez, . Genetic transformation of oil palm-based on selection with hygromycin. pp. 577-587. ISSN 2811-4701
Abstract
The bar gene conferring resistance to the herbicide Basta was used as a selectable marker in oil palm transformation system. However the inefficiency in the selection system was believed to generate transgene escape and high rate of chimerism in oil palm. To overcome this limitation an effective selectable marker for oil palm is required. The aim of this work is to evaluate the use of the hygromycin phosphotransferase (hpt) gene as the selectable marker for generation of stable oil palm transformation via biolistic method and subsequently improve the oil palm transformation efficiency. In this selection approach the embryogenic calli were bombarded with the vectors carrying the hpt and a green fluorescent protein (gfp) reporter genes which were driven by the 2X35S promoter assembled in pBINPLUS and pCAMBIA0380 (pPZP) backbones. Visualisation of GFP spots was observed using Fluorescence Microscope for confirmation of successful deoxyribonucleic acid (DNA) delivery. The calli were then cultured on regeneration medium added with hygromycin at 10 mg litre“1 and consequently reduced to 5 mg litre“1. The presence of transgenes in the bombarded tissue was confirmed by polymerase chain reaction (PCR) amplification of hpt and gfp genes. These results demonstrate the potential of hygromycin as an alternative selection agent for oil palm transformation.
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Abstract
The bar gene conferring resistance to the herbicide Basta was used as a selectable marker in oil palm transformation system. However the inefficiency in the selection system was believed to generate transgene escape and high rate of chimerism in oil palm. To overcome this limitation an effective selectable marker for oil palm is required. The aim of this work is to evaluate the use of the hygromycin phosphotransferase (hpt) gene as the selectable marker for generation of stable oil palm transformation via biolistic method and subsequently improve the oil palm transformation efficiency. In this selection approach the embryogenic calli were bombarded with the vectors carrying the hpt and a green fluorescent protein (gfp) reporter genes which were driven by the 2X35S promoter assembled in pBINPLUS and pCAMBIA0380 (pPZP) backbones. Visualisation of GFP spots was observed using Fluorescence Microscope for confirmation of successful deoxyribonucleic acid (DNA) delivery. The calli were then cultured on regeneration medium added with hygromycin at 10 mg litre“1 and consequently reduced to 5 mg litre“1. The presence of transgenes in the bombarded tissue was confirmed by polymerase chain reaction (PCR) amplification of hpt and gfp genes. These results demonstrate the potential of hygromycin as an alternative selection agent for oil palm transformation.
Additional Metadata
| Item Type: | Article |
|---|---|
| AGROVOC Term: | Oil palm |
| AGROVOC Term: | Hygromycin |
| AGROVOC Term: | Biolistics |
| AGROVOC Term: | plant genetics |
| AGROVOC Term: | Transgenic plants |
| AGROVOC Term: | Plant propagation |
| AGROVOC Term: | Laboratory experimentation |
| AGROVOC Term: | Sustainable agriculture |
| AGROVOC Term: | Innovation |
| AGROVOC Term: | Yield increases |
| Depositing User: | Mr. AFANDI ABDUL MALEK |
| Last Modified: | 24 Apr 2025 00:55 |
| URI: | http://webagris.upm.edu.my/id/eprint/10266 |
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