Development of transformation system for oil palm using GFP as the selectable marker gene

Na'imatulapidah Abdul Majid and Ahmad Parveez Ghulam Kadir (2003) Development of transformation system for oil palm using GFP as the selectable marker gene. [Proceedings Paper]

Green fluorescent protein (GFP) originally isolated from the jellyfish Aequorea victoria has been widely used as a reporter and selectable gene for transformation as it can easily be visualized under blue light in many heterologous organisms. This visual selection is nondestructive and does not need any exogenous substance or antibiotic. It has been transiently and stably expressed in a number of plant species. Transient and long-term expression of the gene in oil palm cultures has been obtained and reported earlier. However the GFP was continuously expressed up to 8 months but the callus failed to proliferate, and died after 8 months. Possibility of toxic effect of GFP in oil palm embryogenic calli was proposed and investigated. Possible toxic effect of GFP has been addressed through co-bombardment with Basta resistant gene construct and bombardment using organelle-targeted GFP plasmids (e.g. plastid, mitochondria and endoplasmic reticulum). About 100 plantlets were directly germinated from GFP-bombarded embryos. Regeneration of a small plantlets from calli derived from GFP-bombarded immature embryos were also obtained. PCR analysis on total DNA isolated from these plantlets was carried out. Results obtained- to date and possible explanation of the effect of GFP in oil palm cultures will be elaborated.

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Abstract

Green fluorescent protein (GFP) originally isolated from the jellyfish Aequorea victoria has been widely used as a reporter and selectable gene for transformation as it can easily be visualized under blue light in many heterologous organisms. This visual selection is nondestructive and does not need any exogenous substance or antibiotic. It has been transiently and stably expressed in a number of plant species. Transient and long-term expression of the gene in oil palm cultures has been obtained and reported earlier. However the GFP was continuously expressed up to 8 months but the callus failed to proliferate, and died after 8 months. Possibility of toxic effect of GFP in oil palm embryogenic calli was proposed and investigated. Possible toxic effect of GFP has been addressed through co-bombardment with Basta resistant gene construct and bombardment using organelle-targeted GFP plasmids (e.g. plastid, mitochondria and endoplasmic reticulum). About 100 plantlets were directly germinated from GFP-bombarded embryos. Regeneration of a small plantlets from calli derived from GFP-bombarded immature embryos were also obtained. PCR analysis on total DNA isolated from these plantlets was carried out. Results obtained- to date and possible explanation of the effect of GFP in oil palm cultures will be elaborated.

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Item Type:	Proceedings Paper
Additional Information:	Available at Perpustakaan Sultan Abdul Samad, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. TP684 P3I61 2003 Call Number
AGROVOC Term:	oil palm > oil palm Prefer using Elaeis guineensisElaeis guineensis
AGROVOC Term:	transgenic plants
AGROVOC Term:	sampling
AGROVOC Term:	data analysis
AGROVOC Term:	plasmid vectors
AGROVOC Term:	polymerase chain reaction > polymerase chain reaction Prefer using PCRPCR
AGROVOC Term:	genetic transformation
AGROVOC Term:	plant biotechnology
AGROVOC Term:	transformation
Geographical Term:	Malaysia
Depositing User:	Nor Hasnita Abdul Samat
Date Deposited:	03 Aug 2024 06:26
Last Modified:	03 Aug 2024 06:26
URI:	http://webagris.upm.edu.my/id/eprint/846

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