Journal of Oil Palm Research Vol.   p.  
DOI: https://doi.org/10.21894/jopr.2020.0111

GENETIC TRANSFORMATION OF OIL PALMBASED ON SELECTION WITH HYGROMYCIN

Author(s): BAHARIAH, B*; RASID, O A*; RAHMAH, A R S*; PARVEEZ, G K A* and MASANI, M Y A*

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.

Keywords: , ,

Author Information
* Malaysian Palm Oil Board, 6 Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor, Malaysia.
E-mail: bahariah@mpob.gov.my


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