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dc.contributor.advisorBurton, Rachel-
dc.contributor.authorHerliana, Lina-
dc.description.abstractMucilage released from Plantago ovata seed (psyllium) has been used for maintaining human health as a dietary fibre supplement. Heteroxylan is the main component, and its substitution affects solubility and viscosity of the end product. However, little is known about genes involved in xylan substitution so phylogenetic and transcript information were used to identify candidate genes in the GT61 and DUF579 families and their functions were tested in the model plant Arabidopsis thaliana. Plantago GT61_7, driven by a seed-coat promoter (ProDP1) was transformed into Arabidopsis using a floral dip and spray method. Ruthenium red staining of wild-type and T2 seeds from multiple independent transgenic lines showed a significant difference in the thickness of the adherent mucilage layer. The difference in mucilage phenotype suggests that GT61_7 may have a role in xylan substitution that affects seed coat adherence. This preliminary result needs to be examined using immunolabeling and monosaccharide analysis. For the DUF579 gene AT1G71690, a genome editing approach was adopted. Three single guide RNAs were designed using online tools and in silico analysis was performed to predict any changes in coding and protein sequences by each guide RNA. To test them in vitro, the CRISPR/Cas9 constructs were successfully delivered to protoplast cells using the Transient Expression in Arabidopsis Mesophyll Protoplast (TEAMP) method. However, an analysis using Tracking of Indels by Decomposition(TIDE) showed no evidence of edits in the DUF569 genomic DNA extracted from the protoplasts. Increasing the transfection efficiency or redesigning the sgRNA could lead to improved CRISPR/Cas9 activity.en
dc.subjectSeed mucilageen
dc.subjectglycosyltransferase (GT61)en
dc.subjectDomain of Unknown Function (DUF) 579en
dc.subjectPlantago ovataen
dc.subjectArabidopsis thalianaen
dc.titleHeterologous Expression and Functional Analysis of Plantago GT61 and DUF579 Genes in Arabidopsis thalianaen
dc.provenanceThis electronic version is made publicly available by the University of Adelaide in accordance with its open access policy for student theses. Copyright in this thesis remains with the author. This thesis may incorporate third party material which has been used by the author pursuant to Fair Dealing exceptions. If you are the owner of any included third party copyright material you wish to be removed from this electronic version, please complete the take down form located at:
dc.description.dissertationThesis (M.Bio.(PB)) -- University of Adelaide, Masters of Biotechnology (Plant Biotechnology), School of Agriculture, Food and Wine, 2017.-
Appears in Collections:School of Agriculture, Food and Wine

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