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Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize
Dissertation (MSc)--University of Pretoria, 2023.
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| Format: | Thesis |
| Language: | English |
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University of Pretoria
2023
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| _version_ | 1867613547778801664 |
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| access_status_str | Open Access |
| author2 | Berger, David Kenneth |
| author_browse | Berger, David Kenneth |
| author_facet | Berger, David Kenneth |
| collection | Thesis |
| dc_rights_str_mv | © 2023 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc)--University of Pretoria, 2023. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/91278 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:37:53.400Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/91278 Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize Berger, David Kenneth carla.buitendag@up.ac.za Theron, Jacques Buitendag, Carla Maize Grey leaf spot Cercospora zeina RNA interference Spray-induced gene silencing (SIGS) UCTD Dissertation (MSc)--University of Pretoria, 2023. Grey leaf spot (GLS) is a devastating fungal foliar disease which affects maize globally. The causal agent for GLS in southern Africa is Cercospora zeina. Current methods to control GLS include chemical fungicides and tilling. However, GLS remains a destructive disease with up to 60% yield losses reported in susceptible hybrids. Advances in plant biotechnology utilize a natural gene silencing mechanism known as RNA interference (RNAi). RNAi is mediated by double-stranded RNA (dsRNA) that targets the expression of genes that share sequence complementarity with the dsRNA. This phenomenon has been adapted in new crop protection strategies such as spray-induced gene silencing (SIGS), which involves the external application of dsRNA that targets pathogenicity genes to combat disease. SIGS is a gene-specific, environmentally friendly, biodegradable, and safe treatment. In a previous study from our research group a dsRNA construct was generated which targets three pathogenicity genes in C. zeina. The in vivo results indicated that this dsRNA construct significantly reduced the fungal cell viability (α = 0.05). In this study, the aim was to test the ability of the dsRNA construct to control GLS disease in planta. Maize leaves were inoculated with C. zeina conidia and treated with the dsRNA construct or a negative dsRNA treatment control, Green fluorescent protein (GFP) dsRNA. In addition, the percentage of germinated conidia over a 24-h period was determined in vitro with light microscopy (LM) and in planta using scanning electron microscopy (SEM). This study demonstrated that the dsRNA construct significantly reduced GLS disease symptoms, compared to the positive disease control (α = 0.05). The GFP dsRNA treatments resulted in similar GLS disease symptoms as the positive disease control. A new dsRNA construct was created based on previous successful SIGS studies in Botrytis cinerea, which had demonstrated efficient disease control using the application of external dsRNA. The new dsRNA construct designed in this study will be evaluated in future studies with the aim to obtain highly effective SIGS results in C. zeina which produce almost no GLS symptoms in maize. This study demonstrates promising results that RNAi technologies such as SIGS could be used to control GLS disease in maize. National Research Foundation Plant Science MSc Unrestricted 2023-07-05T09:40:12Z 2023-07-05T09:40:12Z 2023-09 2023 Dissertation Buitendag, C. 2023, Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize, MSc dissertation, University of Pretoria http://hdl.handle.net/2263/91278 https://doi.org/10.25403/UPresearchdata.22592959.v1 en © 2023 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | Maize Grey leaf spot Cercospora zeina RNA interference Spray-induced gene silencing (SIGS) UCTD Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize |
| title | Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize |
| title_full | Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize |
| title_fullStr | Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize |
| title_full_unstemmed | Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize |
| title_short | Gene silencing mediated by dsRNA reduces grey leaf spot disease in maize |
| title_sort | gene silencing mediated by dsrna reduces grey leaf spot disease in maize |
| topic | Maize Grey leaf spot Cercospora zeina RNA interference Spray-induced gene silencing (SIGS) UCTD |
| url | http://hdl.handle.net/2263/91278 https://doi.org/10.25403/UPresearchdata.22592959.v1 |