Optimisation of the bombardment parameters for transformation and regeneration of stable oil palm transformants
2020
Nurfahisza A. Rahman | Ahmad Parveez Ghulam Kadir | Omar Abdul Rasid | Mat Yunus Abdul Masani
Genetic transformation approaches by using particle bombardment have been an integral part in efforts to improve palm oil quality. The bombardment parameters were previously evaluated to efficiently deliver deoxyribonucleic acid (DNA) during bombardment. However, the evaluation was carried out at transient stage and therefore the results may not be very precise. In order to improve the DNA delivery and regeneration of the oil palm carrying transgenes, a study at stable stage was performed and discussed in this paper. Oil palm embryogenic calli were bombarded at several parameters using pAHC25 plasmid. Selection on media containing 1 mg litre-¹ of bialaphos was initiated after one month of bombardment. Then, the concentration of bialaphos was increased to 2 mg litre-¹ and subsequently to 3 mg litre-¹ to effectively eliminate the regeneration of untransformed cells. Generated shoots were subjected to polymerase chain reaction (PCR) analysis. The best bombardment condition was determined based on the highest number of shoots carrying the transgene. The bombardment at 1100 psi helium pressure with 7.5 cm distance between stopping plate to tissue and one time bombardment using 2.0 μg DNA per bombardment was obtained as the best condition for DNA delivery and also minimised tissue damage. Hence, the results of this study will be useful as a guideline to generate transformed oil palm.
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