Transcriptome analysis of mature seeds of Moringa oleifera and identification of genes related to lipid biosynthesis

Date of Award

2018

Document Type

Thesis

Degree Name

Master of Science in Biology

Department

Biology

First Advisor

Panes, Vivian A., Ph.D.

Abstract

Despite the great potential of Moringa oil to address several health-related problems, there is a scarcity of information on transcriptome level studies to determine the genes that code for enzymes involved in the lipid biosynthesis. Functional annotation of the gene products encoded by the transcript assemblies (Trinity and Oases) from a previous study by Panes et al. 2017 were determined using databases such as NCBI NR, Swissprot, Uniref, and Pfam. Moreover COG classifications, GO assignments and KEGG pathways assignments were also done. Gene expression analysis was done using the FPKM values of the transcripts. PCR and qPCR were utilized to validate selected lipid biosynthesis and reference housekeeping genes of M. oleifera. It was determined that Theobroma cacao is closely related to M. oleifera by having the highest percentage of protein hits for both assemblers. The identification of ACP, KAS III, FAD2, FATB, GPAT, and those coding for oleosin and caleosin will serve as possible gene targets for future crop improvement of M. oleifera. Amplification of selected lipid biosynthesis genes, ACCase and oleosin, and three housekeeping genes GAPDH, RPL1, and EF2 using PCR verified their presence in the transcriptome. Finally, single peaks generated from qPCR melt curve illustrations of GAPDH, RPL1, and EF2 confirmed their presence in M. oleifera seeds. Thus, the transcriptome profile of the mature seed embryo of M. oleifera generated from this study should shed light on its potential as a good source of oil significant for human consumption due to its nutritive properties.

Comments

The B5.B335 2018

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