Citation
Morris, Priscilla Elizabeth
(2019)
Transcriptome analysis of mesocarp tissues in oil palm (Elaeis guineessis jacq.) MPOB-angola dura across various developmental stages.
Masters thesis, Universiti Putra Malaysia.
Abstract
Oil palm (Elaeis guineessis) Malaysian Palm Oil Board (MPOB) Angolan
germplasm materials were evaluated for their potential use in oil palm breeding
programmes. Large fruit characteristic of some Angola dura palms has provided
breeders with a source of favourable traits for introgression into the current
planting materials. With the aim of understanding molecular regulation during
mesocarp development, maturation and ripening of Angola dura palms, efforts
were initiated to profile the expression of transcripts across nine different
developmental stages of mesocarp tissues. The second objective of this study
was to identify differentially expressed genes associated with lipid metabolism,
carbohydrate metabolism, transcription factors and plant hormone metabolism
in mesocarp. The final objective was to profile the expression of selected
WRINKLED1 transcripts across different developmental stages of mesocarp,
endosperm and vegetative tissues. A total of 36,675 genes were identified from
RNA-Sequencing with 24,226 transcripts successfully annotated with the Plant
Reference Sequence Database using BLASTX. Pairwise T-test was performed
using TIGR Multiexperiment Viewer and a total of 21,261 transcripts were
identified as significantly differentially expressed across all the pairwise
comparisons. BLAST2GO analysis assigned 13,996 unigenes with various GO
terms. Transcripts associated with lipid metabolic process were highly
expressed during lag phase preceding the lipid biosynthesis [(10 to 12 Week
After Anthesis (WAA)] and fruit maturation (18 to 20 WAA) stages. Meanwhile,
transcripts linked with carbohydrate metabolic process were up regulated
during transition of fruit developmental stages from 10 to 12 WAA. Further
annotation of the unigenes with KEGG pathway identified 279, 757 and 142 transcripts related to lipid, carbohydrate and hormone metabolisms,
respectively. Additionally, plant RefSeq database annotated 272 transcripts
encoding transcription factors. The most stable reference genes for RT-qPCR
were selected based on RefFinder software. Twenty-seven transcripts from the
RNA-Seq data of nine developmental stages associated with lipid,
carbohydrate, plant hormone metabolism and transcription factors were chosen
for validation of expression levels using Fluidigm 48.48 Real-Time qPCR.
Transcripts such as KAS I, FATA, DGAT, ENR, WRI1 and bZIP showed different
expression profiles in the MPOB-Angola dura as compared to the previously
reported data. WRI1-2 was highly expressed in the mesocarp of MPOB-Angola
dura at the beginning of oil accumulation from 20 WAA to 24 WAA with log2
(fpkm) of 4.7, together with the up-regulation of FA (and TAG biosynthetic
genes In contrast, WRI1-4 was highly expressed in the endosperm of MPOBAngola
dura from 8-15 WAA and decreased at 18 WAA with a fold change of
4.9 which corresponded with the expression profiles of KAS I in the endosperm
tissues. The availability of these transcriptome datasets gives an insight into the
transcriptional mechanisms controlling the Angolan dura fruit development,
maturation and ripening.
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