Optimisation of the bombardment parameters for transformation and regeneration of stable oil palm transformants

Nurfahisza A. Rahman, . and Ahmad Parveez Ghulam Kadir, . and Omar Abdul Rasid, . and Mat Yunus Abdul Masani, . Optimisation of the bombardment parameters for transformation and regeneration of stable oil palm transformants. pp. 696-704. ISSN 1511-2780

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- of bialaphos was initiated after one month of bombardment. Then the concentration of bialaphos was increased to 2 mg litre- and subsequently to 3 mg litre- 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 g 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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Abstract

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- of bialaphos was initiated after one month of bombardment. Then the concentration of bialaphos was increased to 2 mg litre- and subsequently to 3 mg litre- 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 g 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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Item Type:	Article
AGROVOC Term:	Oil palm
AGROVOC Term:	Genetic transformation
AGROVOC Term:	DNA
AGROVOC Term:	Optimization methods
AGROVOC Term:	Polymerase chain reaction
AGROVOC Term:	Shoots
AGROVOC Term:	Transgenic plants
AGROVOC Term:	Transformation (of products)
Depositing User:	Mr. AFANDI ABDUL MALEK
Last Modified:	24 Apr 2025 00:55
URI:	http://webagris.upm.edu.my/id/eprint/9480

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