title: Genetic transformation of Metroxylon sagu (Rottb.) cultures via Agrobacterium-mediated and particle bombardment.
abstract: Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.
authors.affiliation: CRAUN Research Sdn. Bhd., 93055 Kuching, Sarawak, Malaysia.
authors.forename: Evra Raunie
authors.initials: ER
authors.lastname: Ibrahim
authors.affiliation: Genetic Engineering Laboratory, Department of Molecular Biology, Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia ; Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi 6205, Bangladesh.
authors.forename: Md Anowar
authors.identifier: 0000-0003-3967-2544
authors.initials: MA
authors.lastname: Hossain
authors.affiliation: Genetic Engineering Laboratory, Department of Molecular Biology, Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia.
authors.forename: Hairul Azman
authors.identifier: 0000-0002-6700-6138
authors.initials: HA
authors.lastname: Roslan
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D028023:Arecaceae / Q000235:genetics* / Q000254:growth & development; D002146:Bony Callus / Q000196:embryology / Q000254:growth & development; D006046:Gold / Q000737:chemistry; D006371:Helium; D030821:Plants, Genetically Modified / Q000235:genetics*; D011312:Pressure; D013213:Starch / Q000096:biosynthesis; D014170:Transformation, Genetic
chemical_list: D006371:Helium; D006046:Gold; D013213:Starch