ARTICLE IN PRESS

DETERMINATION OF RELIABLE REFERENCE GENES FOR REVERSE TRANSCRIPTION QUANTITATIVE REAL-TIME PCR FROM OIL PALM TRANSCRIPTOMES

NADZIRAH AMIRUDDIN1 ; PEK-LAN CHAN1 ; KUANG-LIM CHAN1 ; PEI-WEN ONG1 ; PRISCILLA ELIZABETH MORRIS1 ; MEILINA ONG-ABDULLAH1 ; SUBHI SITI MASURA1 and ENG-TI LESLIE LOW1*

DOI: https://doi.org/10.21894/jopr.2022.0002
Received: 21 May 2021   Accepted: 2 November 2021   Published Online: 17 January 2022
ABSTRACT

A set of reliable reference genes is essential for accurate quantification and interpretation of gene expression data using reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). In this study, Roche-454 RNA-seq reads from 27 libraries of various oil palm tissues were systematically analysed to identify a set of potential reference genes. Eleven candidate reference genes were identified from the transcriptome data. These genes, together with three oil palm reference genes previously identified for tissue culture samples (PD000380, PD00569, pOP-EA01332) and five classical housekeeping genes [glyceraldehyde-3-phosphate dehydrogenase (GAPDH), NAD5, TUBULIN, UBIQUITIN, ACTIN] were analysed across samples collected from various tissues from mature oil palm (leaf, root, endosperm, mesocarp, female flowers) and stages of tissue culture (non-embryogenic callus, embryogenic callus, polyembryoids, and shoots from polyembryoids). The expression levels of these genes were compared and evaluated using geNorm. Three genes (EgREF_5, EgREF_7 and EgREF_11) were found to be appropriate reference genes for normalising gene expression data from both mature plant and tissue culture samples.

KEYWORDS:


1 Malaysian Palm Oil Board,
6 Persiaran Institusi, Bandar Baru Bangi,
43000 Kajang, Selangor, Malaysia.

* Corresponding author e-mail: lowengti@mpob.gov.my