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Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading

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

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Main Author: Smith, Marthinus Luther
Other Authors: Groenewald, J.-H.
Format: Thesis
Language:English
Published: Stellenbosch : Stellenbosch University 2008
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access_status_str Open Access
author Smith, Marthinus Luther
author2 Groenewald, J.-H.
author_browse Groenewald, J.-H.
Smith, Marthinus Luther
author_facet Groenewald, J.-H.
Smith, Marthinus Luther
author_sort Smith, Marthinus Luther
collection Thesis
dc_rights_str_mv Stellenbosch University
description Thesis (MSc (Genetics. Plant Biotechnology)) --Stellenbosch University, 2008.
format Thesis
id oai:scholar.sun.ac.za:10019.1/3062
institution Stellenbosch University (South Africa)
language English
last_indexed 2026-06-10T12:44:48.111Z
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:10019.1/3062 Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading Smith, Marthinus Luther Groenewald, J.-H. Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics. Institute for Plant Biotechnology. Pyrophosphate Phloem metabolism Phosphotransferase Sugarcane genetics Theses -- Plant biotechnology Dissertations -- Plant biotechnology Plant cells and tissues Vascular system of plants Sugarcane -- Biotechnology Genetics Institute for Plant Biotechnology Thesis (MSc (Genetics. Plant Biotechnology)) --Stellenbosch University, 2008. The main aim of the work presented in this thesis was to further our understanding of the role of Pyrrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) in sugarcane, by specifically investigating its potential contribution to phloem metabolism. PFP activity in sugarcane internodal tissue is inversely correlated to sucrose content across varieties that differ in their sucrose accumulation abilities. This apparent correlation is in contrast to previous studies that suggest PFP plays an insignificant role in metabolism. In the first part of this study an immunological characterisation of the two subunits of sugarcane PFP was conducted to establish whether it differ significantly from other plant species in terms of size and distribution. Both the alpha and beta subunit appears to be approximately sixty kilo Daltons in size and uniform in their relative distribution to each other in the various plant organs of sugarcane. Although the observed alpha subunit size is less than that predicted this could be explained at the hand of post translational modification, in essence the sugarcane PFP subunits appear similar than that described for other plants especially that of tobacco which was employed as a model system later on in this study. The only direct way to investigate PFP’s contribution to phloem metabolism is to alter its activity by recombinant DNA technologies. Therefore, in the second part of the study transformation systems were designed for both the constitutive and phloem specific downand up-regulation of PFP activity. For the down-regulation of activity a post transcriptional gene silencing system, i.e. a complementary strand intron hairpin RNA (ihpRNA) silencing system, was employed. A partial sequence of the PFP-beta subunit was isolated and used in vector construction. For the over-expression the Giardia lamblia PFP gene was used. The model plant tobacco was employed to investigate PFP’s effect on phloem metabolism and transport of assimilate. Transgene insertion was accomplished by means of Agobacterium mediated transformation and tissue specific manipulation of PFP activity was confirmed by in situ activity staining. Masters 2008-11-25T12:10:35Z 2010-06-01T09:05:16Z 2008-11-25T12:10:35Z 2010-06-01T09:05:16Z 2008-12 Thesis http://hdl.handle.net/10019.1/3062 en Stellenbosch University application/pdf Stellenbosch : Stellenbosch University
spellingShingle Pyrophosphate
Phloem metabolism
Phosphotransferase
Sugarcane genetics
Theses -- Plant biotechnology
Dissertations -- Plant biotechnology
Plant cells and tissues
Vascular system of plants
Sugarcane -- Biotechnology
Genetics
Institute for Plant Biotechnology
Smith, Marthinus Luther
Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading
title Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading
title_full Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading
title_fullStr Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading
title_full_unstemmed Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading
title_short Investigating the role of pyrophosphate fructose 6-phosphate 1-phosphotransferase in phloem loading
title_sort investigating the role of pyrophosphate fructose 6 phosphate 1 phosphotransferase in phloem loading
topic Pyrophosphate
Phloem metabolism
Phosphotransferase
Sugarcane genetics
Theses -- Plant biotechnology
Dissertations -- Plant biotechnology
Plant cells and tissues
Vascular system of plants
Sugarcane -- Biotechnology
Genetics
Institute for Plant Biotechnology
url http://hdl.handle.net/10019.1/3062
work_keys_str_mv AT smithmarthinusluther investigatingtheroleofpyrophosphatefructose6phosphate1phosphotransferaseinphloemloading