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https://ptsldigital.ukm.my/jspui/handle/123456789/500081Full metadata record
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | Abdul Munir Abd. Murad, Assoc. Prof. Dr. | - |
| dc.contributor.author | Chan Pek Lan (P42581) | - |
| dc.date.accessioned | 2023-10-13T09:38:07Z | - |
| dc.date.available | 2023-10-13T09:38:07Z | - |
| dc.date.issued | 2017-03-08 | - |
| dc.identifier.other | ukmvital:100115 | - |
| dc.identifier.uri | https://ptsldigital.ukm.my/jspui/handle/123456789/500081 | - |
| dc.description | Micropropagation of oil palm through tissue culture is vital for the generation of superior and uniform elite planting materials. However, one of the major challenges encountered by the oil palm industry is the low callogenesis and embryogenesis rates observed in large scale commercial propagation. In this study, efforts were carried out to identify genes with expression profiles that can differentiate leaf explants producing either embryogenic or non-embryogenic callus. Oil palm cDNA microarrays representing 3891 genes, developed by the Malaysia Palm Oil Board (MPOB), were co-hybridized with cDNA probes of reference tissue and embryo forming or non-embryo forming leaf explants sampled at three time points (Day7, Day14 and Day21) across two tissue culture media treatments (T527 and T694). Tissue culture media T527 successfully produced embryogenic callus followed by embryoids (EMB), whereas media T694 produced non-embryogenic callus. Analysis of the normalized datasets using Significance Analysis of Microarrays (SAM) statistical software resulted in the identification of 77, 115 and 127 significant differentially expressed genes at Day7, Day14 and Day21, respectively. Nine genes from different functional classes were selected for validation via reverse transcription quantitative real-time PCR (RT-qPCR). The expression profiles of all genes observed in RT-qPCR were highly consistent with the cDNA microarray data. One of the genes coding for an early nodulin 93 protein (ENOD93), which was highly expressed at Day7, Day14 and Day21 in leaf explants from media T527, was selected for further analysis. The expression profile of EgENOD93 was also determined from the leaf explants on Day21, 28 and 56 as well as from the callus and EMB. It was observed that EgENOD93 transcripts were up-regulated in the callus obtained from the same media. When the EgENOD93 gene expression analysis was performed across tissue culture lines of different genetic background and media composition, the expression profiles of this transcript at Day21 and Day28 was significantly differential between leaf explants and could be used to predict those with embryogenic potential. The full-length sequence of EgENOD93 was then isolated using 5'-RACE. Sequence analysis showed that the gene comprised of a 61 bp 5'-untranslated region (UTR), a 300 bp open reading frame and a 229 bp 3'-UTR. Phylogenetic analysis using MEGA7 revealed that the oil palm EgENOD93 was closely related to ENOD93 of Phoenix dactylifera and Potamogeton distinctus. Furthermore, cis-acting elements such as endosperm expression, abscisic acid responsiveness, methyl jasmonate responsiveness, defense and stress responsiveness were identified in the upstream region of the EgENOD93 promoter. In situ RNA hybridization studies confirmed the localization of the EgENOD93 transcript in the embryogenic cells. Functional characterization of the putative orthologous Medicago truncatula gene, executed via the RNAi approach, provided additional evidence that ENOD93 is essential for somatic embryogenesis. Interference in the expression of M. truncatula ENOD93 resulted in the formation of much lower numbers of somatic embryos as compared to the control. This study enhances the understanding on the molecular mechanisms associated with early developmental stages of tissue culture and reveals that EgENOD93 is suitable to be developed as a marker to predict the potential of leaf explants to successfully produce embryogenic callus.,Certification of Master's/Doctoral Thesis" is not available | - |
| dc.language.iso | eng | - |
| dc.publisher | UKM, Bangi | - |
| dc.relation | Faculty of Science and Technology / Fakulti Sains dan Teknologi | - |
| dc.rights | UKM | - |
| dc.subject | Tissue culture | - |
| dc.subject | Micropropagation | - |
| dc.subject | Oil palm | - |
| dc.title | Isolation and characterization of genes associated with somatic embryogenesis in oil palm (Elaeis guineensis) | - |
| dc.type | Theses | - |
| dc.format.pages | 198 | - |
| dc.identifier.callno | SB299.P3C433 2017 tesis | - |
| Appears in Collections: | Faculty of Science and Technology / Fakulti Sains dan Teknologi | |
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|---|---|---|---|---|
| ukmvital_100115+SOURCE1+SOURCE1.0.PDF Restricted Access | 196.79 kB | Adobe PDF |  View/Open |
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