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Studies on transgenic tobacco plants containing geminiviral DNA

The ability to transform plant genomes with foreign genetic material by means of the Agrobacterium tumefaciens Ti plasmid system has made it possible to test whether plant resistance to viruses can be increased by incorporating viral nucleotide sequences into the nuclear genome of the host. As part...

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Main Author: Babaya, Michelle
Other Authors: Bradfield, Graham
Format: Thesis
Language:English
Published: Faculty of Science 2026
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access_status_str Open Access
author Babaya, Michelle
author2 Bradfield, Graham
author_browse Babaya, Michelle
Bradfield, Graham
author_facet Bradfield, Graham
Babaya, Michelle
author_sort Babaya, Michelle
collection Thesis
description The ability to transform plant genomes with foreign genetic material by means of the Agrobacterium tumefaciens Ti plasmid system has made it possible to test whether plant resistance to viruses can be increased by incorporating viral nucleotide sequences into the nuclear genome of the host. As part of a model system designed to explore the feasibility of making cereals resistant to maize streak virus (MSV), tobacco plants were to be transformed with one or more genome-equivalents (multimers) of Port Elizabeth (PE) and Nigerian (N) isolates of (MSV), and analysed for evidence of expression of the viral coat protein. The constructs were mobilized into A. tumefaciens [C58C1Rif (pGV2260)] and introduced into tobacco by co-cultivation with leaf discs. Putatively transgenic kanamycin-resistant calli were tested for the presence of the plasmid vector marker enzyme, neomycin phosphotransferase (NPT-II). Southern hybridization analysis was used to establish the integration of MSV DNA into tobacco. Expression of the MSV coat protein was tested for by Western blotting and enzyme-linked immunosorbent . assay ( ELISA) techniques. Transformed calli all expressed NPT-II activity and contained integrated MSV DNA, but no coat protein could be detected serologically. A minor part of the project involved the study of a new strain of MSV isolated from the Koedoeskop area in South Africa which produced severe disease symptoms in maize. The genomic DNA was purified, the genome cloned into pUC19 and mapped. The restriction enzyme cleavage map of the new isolate, MSV-KoeI, was compared to. those of 8 other MSV isolates. Similarities and differences between these maps are discussed.
format Thesis
id oai:open.uct.ac.za:11427/43028
institution University of Cape Town (South Africa)
language eng
license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
publishDate 2026
publishDateRange 2026
publishDateSort 2026
publisher Faculty of Science
publisherStr Faculty of Science
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source_str UCTD — University of Cape Town Open Access Repository
spelling oai:open.uct.ac.za:11427/43028 Studies on transgenic tobacco plants containing geminiviral DNA Babaya, Michelle Bradfield, Graham Science The ability to transform plant genomes with foreign genetic material by means of the Agrobacterium tumefaciens Ti plasmid system has made it possible to test whether plant resistance to viruses can be increased by incorporating viral nucleotide sequences into the nuclear genome of the host. As part of a model system designed to explore the feasibility of making cereals resistant to maize streak virus (MSV), tobacco plants were to be transformed with one or more genome-equivalents (multimers) of Port Elizabeth (PE) and Nigerian (N) isolates of (MSV), and analysed for evidence of expression of the viral coat protein. The constructs were mobilized into A. tumefaciens [C58C1Rif (pGV2260)] and introduced into tobacco by co-cultivation with leaf discs. Putatively transgenic kanamycin-resistant calli were tested for the presence of the plasmid vector marker enzyme, neomycin phosphotransferase (NPT-II). Southern hybridization analysis was used to establish the integration of MSV DNA into tobacco. Expression of the MSV coat protein was tested for by Western blotting and enzyme-linked immunosorbent . assay ( ELISA) techniques. Transformed calli all expressed NPT-II activity and contained integrated MSV DNA, but no coat protein could be detected serologically. A minor part of the project involved the study of a new strain of MSV isolated from the Koedoeskop area in South Africa which produced severe disease symptoms in maize. The genomic DNA was purified, the genome cloned into pUC19 and mapped. The restriction enzyme cleavage map of the new isolate, MSV-KoeI, was compared to. those of 8 other MSV isolates. Similarities and differences between these maps are discussed. 2026-03-19T13:13:15Z 2026-03-19T13:13:15Z 1990 2024-07-19T13:07:25Z Thesis / Dissertation Masters MSc http://hdl.handle.net/11427/43028 eng application/pdf Faculty of Science University of Cape Town
spellingShingle Science
Babaya, Michelle
Studies on transgenic tobacco plants containing geminiviral DNA
thesis_degree_str Master's
title Studies on transgenic tobacco plants containing geminiviral DNA
title_full Studies on transgenic tobacco plants containing geminiviral DNA
title_fullStr Studies on transgenic tobacco plants containing geminiviral DNA
title_full_unstemmed Studies on transgenic tobacco plants containing geminiviral DNA
title_short Studies on transgenic tobacco plants containing geminiviral DNA
title_sort studies on transgenic tobacco plants containing geminiviral dna
topic Science
url http://hdl.handle.net/11427/43028
work_keys_str_mv AT babayamichelle studiesontransgenictobaccoplantscontaininggeminiviraldna