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Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides

Maize is a socially and economically important crop in Africa (and worldwide) that is severely affected by many fungal pathogens. The pathogen Fusarium verticillioides causes Fusarium ear rot in maize, a disease that greatly reduces quantity and quality of annual maize yields. The pathogen produces...

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Main Author: Veenstra, Amy
Other Authors: Murray, Shane L
Format: Thesis
Language:English
Published: Department of Molecular and Cell Biology 2018
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access_status_str Open Access
author Veenstra, Amy
author2 Murray, Shane L
author_browse Murray, Shane L
Veenstra, Amy
author_facet Murray, Shane L
Veenstra, Amy
author_sort Veenstra, Amy
collection Thesis
description Maize is a socially and economically important crop in Africa (and worldwide) that is severely affected by many fungal pathogens. The pathogen Fusarium verticillioides causes Fusarium ear rot in maize, a disease that greatly reduces quantity and quality of annual maize yields. The pathogen produces mycotoxins called fumonisins, which have been linked to adverse health effects in both humans and animals. Maize produces terpenoid phytoalexins, which are antimicrobial compounds that directly reduce the growth of many fungal pathogens including F. verticillioides. Two families of maize phytoalexins, termed kauralexins and zealexins, have been characterized. Key genes putatively involved in the biosynthetic pathway of these phytoalexins have been identified from the rice model and subsequent studies on maize. This research aimed to evaluate the correlation between phytoalexin accumulation and fungal growth in diverse southern African maize lines in response to F. verticillioides inoculation. Maize lines were inoculated with F. verticillioides using a seed soak inoculation method and grown in vitro for up to two weeks. The harvested tissue was analysed for fungal growth using quantitative PCR, putative phytoalexin biosynthetic gene expression using RT-qPCR and phytoalexin accumulation using gas-chromatography mass spectrometry. Furthermore, an endophyte growing in one of the maize lines was isolated and identified as Trichoderma asperellum. Trichoderma spp. are used as biocontrol agents against many fungal pathogens, although research on the specific antagonistic effect of T. asperellum on F. verticillioides is limited. Phytoalexin accumulation in maize containing endophytic T. asperellum was compared to maize inoculated with F. verticillioides. In vitro competition assays were performed to analyse the antagonistic effect of T. asprellum on F. verticillioides. Results from this study show that inoculation of maize lines with F. verticillioides induces the accumulation of total phytoalexins, and more specifically the accumulation of total kauralexins. Putative phytoalexin biosynthetic genes are also up-regulated in response to inoculation. Maize growing with a T. asperellum endophyte accumulated phytoalexins to the same levels as F. verticillioides, suggesting that T. asperellum induces a defence response that 'primes' the plant for further infection. In vitro competition assays between F. verticillioides and T. asperellum showed that T. asperellum significantly inhibits F. verticillioides growth. These results will aid in the identification of maize lines that can be bred with increased resistance to F. verticillioides with the goal to reduce F. verticillioides incidence in southern Africa. Furthermore, analysis of the efficacy of T. asperellum as an antagonist against F. verticillioides may provide another method for disease reduction in the field.
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license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
publishDate 2018
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spelling oai:open.uct.ac.za:11427/27095 Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides Veenstra, Amy Murray, Shane L Rafudeen, Suhail Molecular and Cell Biology Maize is a socially and economically important crop in Africa (and worldwide) that is severely affected by many fungal pathogens. The pathogen Fusarium verticillioides causes Fusarium ear rot in maize, a disease that greatly reduces quantity and quality of annual maize yields. The pathogen produces mycotoxins called fumonisins, which have been linked to adverse health effects in both humans and animals. Maize produces terpenoid phytoalexins, which are antimicrobial compounds that directly reduce the growth of many fungal pathogens including F. verticillioides. Two families of maize phytoalexins, termed kauralexins and zealexins, have been characterized. Key genes putatively involved in the biosynthetic pathway of these phytoalexins have been identified from the rice model and subsequent studies on maize. This research aimed to evaluate the correlation between phytoalexin accumulation and fungal growth in diverse southern African maize lines in response to F. verticillioides inoculation. Maize lines were inoculated with F. verticillioides using a seed soak inoculation method and grown in vitro for up to two weeks. The harvested tissue was analysed for fungal growth using quantitative PCR, putative phytoalexin biosynthetic gene expression using RT-qPCR and phytoalexin accumulation using gas-chromatography mass spectrometry. Furthermore, an endophyte growing in one of the maize lines was isolated and identified as Trichoderma asperellum. Trichoderma spp. are used as biocontrol agents against many fungal pathogens, although research on the specific antagonistic effect of T. asperellum on F. verticillioides is limited. Phytoalexin accumulation in maize containing endophytic T. asperellum was compared to maize inoculated with F. verticillioides. In vitro competition assays were performed to analyse the antagonistic effect of T. asprellum on F. verticillioides. Results from this study show that inoculation of maize lines with F. verticillioides induces the accumulation of total phytoalexins, and more specifically the accumulation of total kauralexins. Putative phytoalexin biosynthetic genes are also up-regulated in response to inoculation. Maize growing with a T. asperellum endophyte accumulated phytoalexins to the same levels as F. verticillioides, suggesting that T. asperellum induces a defence response that 'primes' the plant for further infection. In vitro competition assays between F. verticillioides and T. asperellum showed that T. asperellum significantly inhibits F. verticillioides growth. These results will aid in the identification of maize lines that can be bred with increased resistance to F. verticillioides with the goal to reduce F. verticillioides incidence in southern Africa. Furthermore, analysis of the efficacy of T. asperellum as an antagonist against F. verticillioides may provide another method for disease reduction in the field. 2018-01-30T10:24:59Z 2018-01-30T10:24:59Z 2017 Master Thesis Masters MSc http://hdl.handle.net/11427/27095 eng application/pdf Department of Molecular and Cell Biology Faculty of Science University of Cape Town
spellingShingle Molecular and Cell Biology
Veenstra, Amy
Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides
thesis_degree_str Master's
title Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides
title_full Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides
title_fullStr Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides
title_full_unstemmed Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides
title_short Evaluation of southern African maize germplasm for phytoalexin accumulation following inoculation by Fusarium verticillioides
title_sort evaluation of southern african maize germplasm for phytoalexin accumulation following inoculation by fusarium verticillioides
topic Molecular and Cell Biology
url http://hdl.handle.net/11427/27095
work_keys_str_mv AT veenstraamy evaluationofsouthernafricanmaizegermplasmforphytoalexinaccumulationfollowinginoculationbyfusariumverticillioides