Journal of Oil Palm Research Vol. 27 (4) December 2015, p. 306-314

A CHLOROPLAST DNA (cpDNA) EXTRACTION PROTOCOL FOR DIVERSITY ANALYSIS OF OIL PALM (Elaeis spp.)

HO, CARL MIEW*; OOI, LESLIE C L**; TAN, S G*; HO, C L* and SINGH, R**

ABSTRACT

Oil palm chloroplast is believed to be maternally inherited, making investigation of the chloroplast diversity an interesting endeavour. This article describes a method for extracting enriched oil palm chloroplast DNA (cpDNA) done on six palms of different origins from Angola, Nigeria, Ghana, Madagascar and Surinam. Restriction enzyme digestion was used to evaluate the successful extraction of the oil palm cpDNA. The use of a mitochondrial DNA-specific universal primer revealed that most of the cpDNA were free from mitochondrial DNA contamination. Three chloroplast-specific universal primers were also used to evaluate the cpDNA. Their amplicons were cloned and sequenced to confirm that the cpDNA was indeed amplified.
A search against the public databases further confirmed that the primers amplified sequences of the Elaeis guineensis Jacq. chloroplast genome. Two of them gave consistent amplification when tested on cpDNA from the Angolan, Nigerian, Ghanian, Madagascan and Surinam palms.

KEYWORDS:


* Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.

** Malaysian Palm Oil Board, 6 Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor, Malaysia. E-mail: leslie@mpob.gov.my