Citation
Safiza M., . and Abrizah O., . and Siti Nor Akmar A., . and Mohamad Arif A.M., . and Khairul Anuar R., . and Mahadzir J., . (2007) Confirmation of activity of oil palm stearoyl-acyl carrier protein ACP desaturase by complementation study in Arabidopsis thaliana. [Proceedings Paper]
Abstract
Stearoyl-ACP desaturase SAD is an important fatty acid biosynthetic enzyme which utilizes saturated stearoyl-ACP C18:0-ACP produced by fatty acid synthase as a substrate to produce oleoyl-ACP C18:1-ACP. It is a soluble desaturase which catalyses the desaturation process in the chloroplast stroma. An expression vector pDESCB302-3 was constructed where the mature coding region of pOPSN 19 that codes for SAD 1 was inserted into a binary vector pCB302-3 and placed under the control of the cauliflower mosaic virus CaMV 355 promoter. This binary vector contains biolaphos tolerance gene bar which encodes phosphinothricin acetyltransferase and has been the most widely used selective marker especially for the transformation of cereals. The recombinant plasmid was introduced into wild type Arabidopsis thaliana plants and Arabidopsis lines defective in the stearoyl-ACP desaturase gene i.e. Columbia COL-03 and a fab2FAB-03 via floral dip method with Agrobacterium tumefaciens C58 carrying the recombinant vector. Selection and screening of the transgenic plants was carried out on Murashige and Skoog MS media agar supplemented with Basta. Total fatty acid of the transgenic Arabidopsis thaliana leaves harbouring the above construct was extracted and the fatty acid composition was analyzed using Gas Chromatography. The amount of C18:0 in the transgenic COL-03 leaves decreased to 0.27mol from 2.74mol observed in wild type control plants while the analysis of FAB-03 also shows decreased levels of C18:0 from 7.59mol control to 1.01 mol. Possible explanations for the above observation will be discussed.
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Abstract
Stearoyl-ACP desaturase SAD is an important fatty acid biosynthetic enzyme which utilizes saturated stearoyl-ACP C18:0-ACP produced by fatty acid synthase as a substrate to produce oleoyl-ACP C18:1-ACP. It is a soluble desaturase which catalyses the desaturation process in the chloroplast stroma. An expression vector pDESCB302-3 was constructed where the mature coding region of pOPSN 19 that codes for SAD 1 was inserted into a binary vector pCB302-3 and placed under the control of the cauliflower mosaic virus CaMV 355 promoter. This binary vector contains biolaphos tolerance gene bar which encodes phosphinothricin acetyltransferase and has been the most widely used selective marker especially for the transformation of cereals. The recombinant plasmid was introduced into wild type Arabidopsis thaliana plants and Arabidopsis lines defective in the stearoyl-ACP desaturase gene i.e. Columbia COL-03 and a fab2FAB-03 via floral dip method with Agrobacterium tumefaciens C58 carrying the recombinant vector. Selection and screening of the transgenic plants was carried out on Murashige and Skoog MS media agar supplemented with Basta. Total fatty acid of the transgenic Arabidopsis thaliana leaves harbouring the above construct was extracted and the fatty acid composition was analyzed using Gas Chromatography. The amount of C18:0 in the transgenic COL-03 leaves decreased to 0.27mol from 2.74mol observed in wild type control plants while the analysis of FAB-03 also shows decreased levels of C18:0 from 7.59mol control to 1.01 mol. Possible explanations for the above observation will be discussed.
Additional Metadata
| Item Type: | Proceedings Paper |
|---|---|
| AGROVOC Term: | ELAEIS GUINEENSIS |
| AGROVOC Term: | OIL PALMS |
| AGROVOC Term: | ACYLATION |
| AGROVOC Term: | BINDING PROTEINS |
| AGROVOC Term: | ARABIDOPSIS THALIANA |
| AGROVOC Term: | SEEDS |
| AGROVOC Term: | STERILIZATION |
| AGROVOC Term: | FATTY ACIDS |
| AGROVOC Term: | TRANSGENICS |
| AGROVOC Term: | TRANSGENIC PLANTS |
| Geographical Term: | MALAYSIA |
| Depositing User: | Ms. Norfaezah Khomsan |
| Last Modified: | 24 Apr 2025 05:27 |
| URI: | http://webagris.upm.edu.my/id/eprint/15727 |
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