AsPac J. Mol. Biol. Biotechnol., Vol. 15 (1), 2007 1 Asia Paciic Journal of Molecular Biology and Biotechnology, 2007 Vol. 15 (1) : 1-8 *Author for Correspondence. Mailing address: Advanced Biotechnology and Breeding Center, Biological Research Division,Malaysian Palm Oil Board (MPOB), P.O. Box 10620, 50720 Kuala Lumpur, Malaysia. E-mail: parveez@mpob.gov.my Evaluation of Green Fluorescence Protein (GFP) as a Selectable Marker for Oil Palm Transformation via Transient Expression Na’imatulapidah Abdul Majid and Ghulam Kadir Ahmad Parveez* Advanced Biotechnology and Breeding Center, Biological Research Division, Malaysian Palm Oil Board (MPOB), P.O. Box 10620, 50720 Kuala Lumpur, Malaysia. Received 29 June2006 / Accepted 8 December 2006 Abstract. Green luorescent protein (GFP) can be easily visualized under UV/blue light without any additional substrate or co-factor. Its assay is also non-destructive. Therefore, it has been widely used to monitor transgene expression and protein localization in variety of cells and organisms. To evaluate the effectiveness and practicality of GFP as a reporter and selectable marker for oil palm transformation, transient expression of GFP genes in oil palm embryogenic calli and immature embryos were analyzed after bombardment with different types (version) of GFP genes and driven by different constitutive promoters. Some of the GFP genes used were targeted to speciic organelles within the cell, namely mitochondria, plastid and endoplasmic reticulum (ER). For transformation using non-targeted GFP genes, plasmid p35S.TYG.GFP showed the highest number of green luorescent spots, followed by pHBT-sGFPS65T and pGEM.Ubi-sGFPS65T. The results demonstrated that the sGFPS65T gene version is the most active in oil palm. When organelle targeted GFP genes were transformed, localization of the genes into speciic organelle was clearly demonstrated using laser scanning confocal microscopy. Using the organelle targeted GFP genes; the sGFPS65T gene version was again demonstrated to be the most active in oil palm cells. Long term transient expression of the GFP genes was also evaluated and is discussed. Keywords. Transformation, green luorescence protein (GFP), selectable marker, transient expression, oil palm INTRODUCTION Plant genetic engineering is progressing very rapidly since the irst success of introducing a foreign gene into a plant via Agrobacterium tumefaciens (Fraley, et al., 1983). Since then the number of transgenic plants produced has increased exponentially. The International Service for Acquisition of Agri-biotech Applications (ISAAA) has recently summarized that the area commercially planted with transgenic plants worldwide has increased almost 53 fold, from 1.7 million hectares in 1996 to 90 million in 2005 (James, 2005). Through genetic engineering, agronomic traits of a particular plant can be improved and furthermore, production of value added products and nutrients can also be obtained. Genetic engineering reduces the time required for introducing a novel trait into plants as compared to conventional breeding. It was postulated for oil palm; up to 80% reduction in time could be achieved for introducing a novel trait through genetic engineering. Furthermore, transgenic plants can be used as bioreactors or bio-factories for producing novel products, such as pharmaceuticals, continuously. Oil palm is one of the world’s main sources of vegetable oils and fats. Its production per planted area is 3 times and 10 times higher than coconut and soybean, respectively. In Malaysia, crude palm oil production is increasing annually; from 14 million tonnes in 2004 it has increased to 15 million tonnes in 2005, an increase of 7.1% (Basri, 2006). Similarly, the total export volume of oil palm products, such as palm oil, palm kernel oil and oleochemicals has increased by 7.3%, to 18.62 million tonnes in 2005 as compared to 17.36 million tonnes in 2004 (Basri, 2006). It is expected that the rate of production and export of oil palm products will continue to progress in the years to come and it will remain as one of the major sources of vegetable oils and fats to feed the