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Analysis of intermediate carbon metabolism in strawberry plants

Thesis (MSc (Genetics. Institute for Plant Biotechnology)--Stellenbosch University, 2008.

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Main Author: Basson, Carin Elizabeth
Other Authors: Groenewald, J.-H.
Format: Thesis
Language:English
Published: Stellenbosch : Stellenbosch University 2008
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access_status_str Open Access
author Basson, Carin Elizabeth
author2 Groenewald, J.-H.
author_browse Basson, Carin Elizabeth
Groenewald, J.-H.
author_facet Groenewald, J.-H.
Basson, Carin Elizabeth
author_sort Basson, Carin Elizabeth
collection Thesis
dc_rights_str_mv Stellenbosch University
description Thesis (MSc (Genetics. Institute for Plant Biotechnology)--Stellenbosch University, 2008.
format Thesis
id oai:scholar.sun.ac.za:10019.1/2295
institution Stellenbosch University (South Africa)
language English
last_indexed 2026-06-10T12:42:04.592Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository
publishDate 2008
publishDateRange 2008
publishDateSort 2008
publisher Stellenbosch : Stellenbosch University
publisherStr Stellenbosch : Stellenbosch University
record_format dspace
source_str SUNScholar — Stellenbosch University Repository
spelling oai:scholar.sun.ac.za:10019.1/2295 Analysis of intermediate carbon metabolism in strawberry plants Basson, Carin Elizabeth Groenewald, J.-H. Bauer, R. Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics. Institute for Plant Biotechnology. Carbon metabolism of strawberries Biotechnology Pyrophosphate Carbon partitioning in plants Theses -- Plant biotechnology Dissertations -- Plant biotechnology Strawberries -- Metabolism Genetics Institute for Plant Biotechnology Thesis (MSc (Genetics. Institute for Plant Biotechnology)--Stellenbosch University, 2008. Strawberry (Fragaria x ananassa) fruit quality is largely determined by the relative amounts of sugars and organic acids present, as well as soluble solid content. This study had three components: 1) Characterisation of cytosolic carbohydrate metabolism and carbon partitioning to sugars and organic acids in two commercial varieties, 2) analysis of transgenic strawberry fruit with increased pyrophosphate: D-fructose-6-phosphate 1-phosphotransferase (PFP) activity and 3) analysis of transgenic strawberry fruit with increased ß-fructosidase (invertase) activity in either cytosol or apoplast. Analyses of transgenic strawberry may inform similar attempts in grape berries. Festival and Ventana, two popular commercial strawberry cultivars in South Africa, were fairly similar with respect to sugar and organic acid content. Twelve cytosolic enzymes were investigated. Temporal differences in maximum catalytic activity were observed for invertase, PFP, pyruvate kinase and ADP-glucose pyrophosphorylase (AGPase). Invertase, PFP and AGPase activity also differed between the cultivars. One enzyme, SuSy, could not be analysed effectively, due to the purification method employed. These analyses established methodology for the analysis of transgenic berries. Constructs were designed to constituitively express Giardia lamblia PFP (GL-PFP), or to express Saccharomyces cerevisiae invertase (SCI) in a fruit-specific manner. A second invertase construct was designed to target SCI to the apoplast. Strawberry (cv. Selekta) was transformed and the presence of each transgene confirmed by PCR. Untransformed Selekta was used as control in both transgenic studies. Transgenic lines were selected based on GL-PFP activity in leaves and total PFP activity in ripe fruit. Sugar and organic acid content of ripe berries with high PFP activity was determined. Although berries displayed marked changes in sugar composition, the total sugar content was similar to controls, in all except one line. Organic acid content was decreased, leading to a clear reduction in organic acid-to-sugar ratio. This points to a gluconeogenic role for PFP in strawberry fruit. Transgenic berries were screened for SCI activity. Berries containing untargeted SCI exhibited total invertase activity similar to controls and were not analysed further. Berries with apoplasttargeted SCI displayed three-fold increases in invertase activity compared to controls. Total sugar content was reduced and exhibited reduced sucrose content relative to hexoses. Despite the effect of increased invertase activity on metabolites, maximum catalytic activity of enzymes involved in cytosolic sucrose, hexose and organic acid metabolism were unchanged. Transgenic plants selected in these studies were subsequently vegetatively replicated and future work will include immature fruit. Masters 2008-11-24T09:50:35Z 2010-06-01T08:45:23Z 2008-11-24T09:50:35Z 2010-06-01T08:45:23Z 2008-12 Thesis http://hdl.handle.net/10019.1/2295 en Stellenbosch University application/pdf Stellenbosch : Stellenbosch University
spellingShingle Carbon metabolism of strawberries
Biotechnology
Pyrophosphate
Carbon partitioning in plants
Theses -- Plant biotechnology
Dissertations -- Plant biotechnology
Strawberries -- Metabolism
Genetics
Institute for Plant Biotechnology
Basson, Carin Elizabeth
Analysis of intermediate carbon metabolism in strawberry plants
title Analysis of intermediate carbon metabolism in strawberry plants
title_full Analysis of intermediate carbon metabolism in strawberry plants
title_fullStr Analysis of intermediate carbon metabolism in strawberry plants
title_full_unstemmed Analysis of intermediate carbon metabolism in strawberry plants
title_short Analysis of intermediate carbon metabolism in strawberry plants
title_sort analysis of intermediate carbon metabolism in strawberry plants
topic Carbon metabolism of strawberries
Biotechnology
Pyrophosphate
Carbon partitioning in plants
Theses -- Plant biotechnology
Dissertations -- Plant biotechnology
Strawberries -- Metabolism
Genetics
Institute for Plant Biotechnology
url http://hdl.handle.net/10019.1/2295
work_keys_str_mv AT bassoncarinelizabeth analysisofintermediatecarbonmetabolisminstrawberryplants