Alternative approach to controlling citrus black spot disease
- Authors: Thabede, Jahman Thabo
- Date: 2021-04
- Subjects: To be added
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/178569 , vital:42951
- Description: Access restricted until April 2022. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2021
- Full Text:
- Authors: Thabede, Jahman Thabo
- Date: 2021-04
- Subjects: To be added
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/178569 , vital:42951
- Description: Access restricted until April 2022. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2021
- Full Text:
Arbuscular mycorrhizal fungi as a bio-indicator of soil health under agricultural management practices in South Africa
- Authors: Sekgota, Wendy Maphefo
- Date: 2019
- Subjects: Soils -- Quality -- South Africa , Soil fertility -- South Africa , Fungi in agriculture -- South Africa , Mycorrhizal fungi , Vesicular-arbuscular mycorrhizas , Fungi -- Spores , Soils -- Agricultural chemical content
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72161 , vital:30011
- Description: This study investigated the activity of arbuscular mycorrhizal (AM) fungi as a potential biological indicator of soil health under conventional and conservation agricultural management in South Africa. An experimental trial consisting of three replicates plots under conventional and reduced tillage subdivided into twelve treatments of six crops and two fertilizer inputs was assessed over four growing seasons for various AM fungal parameters such as spore density, most probable number (MPN) of propagules percentage root colonisation and easily extractable glomalin (EEG). Cropping combinations were maize monoculture; maize soybean rotation; maize cowpea rotation; maize cowpea intercropping; maize oats intercropping and maize vetch intercropping. Resident AM fungal spore numbers and EEG protein levels were very low and no root colonization was recorded in the first two growing seasons. These findings prompted the need for the inoculation of the study site in the third growing season with a commercial AM fungal product (MycorootTM). Spore numbers, EEG concentrations and percentage root colonisation increased 8 weeks after inoculation but were significantly reduced in the fourth growing season that was not inoculated. MPN infectivity increased with inoculation particularly under conventional tillage and maize monoculture. Resident spore taxa were morphologically identified into three genera Gigaspora, Scutellospora, and Glomus. For the first two growing seasons, the maize roots were heavily colonized by a pathogenic fungus after mycorrhizal inoculation no evidence of pathogenic fungi was observed. In the fourth growing season which did not receive inoculation, root colonization started to decline. Reduced tillage, high fertilizer input combined with maize cowpea rotation (MC) and maize hairy vetch intercropping (Mv) had a significant effect (P = 0.01) on AM fungal spore numbers. Cropping systems and high fertilizer input had a significant effect on EEG concentrations in the second growing season. Overall, fertilizer application and crop type had implications for mycorrhizal activity. The soil health status in this study site was deemed low as measured by the impaired mycorrhizal activity due to agricultural management practices. Field inoculation combined with classical and molecular tools could provide a more realistic assessment of the effect of agricultural management practices on AM fungi as potential bioindicators of soil health. Therefore, AM fungi could be used as bioindicators of soil health under agricultural management practices in South African soil conditions.
- Full Text:
- Authors: Sekgota, Wendy Maphefo
- Date: 2019
- Subjects: Soils -- Quality -- South Africa , Soil fertility -- South Africa , Fungi in agriculture -- South Africa , Mycorrhizal fungi , Vesicular-arbuscular mycorrhizas , Fungi -- Spores , Soils -- Agricultural chemical content
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72161 , vital:30011
- Description: This study investigated the activity of arbuscular mycorrhizal (AM) fungi as a potential biological indicator of soil health under conventional and conservation agricultural management in South Africa. An experimental trial consisting of three replicates plots under conventional and reduced tillage subdivided into twelve treatments of six crops and two fertilizer inputs was assessed over four growing seasons for various AM fungal parameters such as spore density, most probable number (MPN) of propagules percentage root colonisation and easily extractable glomalin (EEG). Cropping combinations were maize monoculture; maize soybean rotation; maize cowpea rotation; maize cowpea intercropping; maize oats intercropping and maize vetch intercropping. Resident AM fungal spore numbers and EEG protein levels were very low and no root colonization was recorded in the first two growing seasons. These findings prompted the need for the inoculation of the study site in the third growing season with a commercial AM fungal product (MycorootTM). Spore numbers, EEG concentrations and percentage root colonisation increased 8 weeks after inoculation but were significantly reduced in the fourth growing season that was not inoculated. MPN infectivity increased with inoculation particularly under conventional tillage and maize monoculture. Resident spore taxa were morphologically identified into three genera Gigaspora, Scutellospora, and Glomus. For the first two growing seasons, the maize roots were heavily colonized by a pathogenic fungus after mycorrhizal inoculation no evidence of pathogenic fungi was observed. In the fourth growing season which did not receive inoculation, root colonization started to decline. Reduced tillage, high fertilizer input combined with maize cowpea rotation (MC) and maize hairy vetch intercropping (Mv) had a significant effect (P = 0.01) on AM fungal spore numbers. Cropping systems and high fertilizer input had a significant effect on EEG concentrations in the second growing season. Overall, fertilizer application and crop type had implications for mycorrhizal activity. The soil health status in this study site was deemed low as measured by the impaired mycorrhizal activity due to agricultural management practices. Field inoculation combined with classical and molecular tools could provide a more realistic assessment of the effect of agricultural management practices on AM fungi as potential bioindicators of soil health. Therefore, AM fungi could be used as bioindicators of soil health under agricultural management practices in South African soil conditions.
- Full Text:
Biological properties and interactions of Kalaharituber pfeilii
- Authors: Krele, Viwe
- Date: 2019
- Subjects: Kalaharituber pfeilii , Pezizales -- South Africa , Desert plants -- South Africa , Truffle culture -- South Africa , Plant biochemical genetics , Enzymes -- Analysis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72257 , vital:30022
- Description: Dessert truffles are seasonal macro fungi and have been identified in several parts of the world including South Africa. The first part of the present study dealt with the assessment of the biologically active compounds of the Kalahari truffles found in the Northern Cape of South Africa. Truffles extracts (methanol, ethanol, aqueous) were investigated for their antimicrobial properties towards Gram-positive and Gram-negative bacteria. The results demonstrated that the truffle extracts tested had no inhibitory effects against the bacterial isolates. The truffle mycelial growth was also noted to be ineffective against the selected bacteria. The bacteria tested in the present study showed some antagonistic effects against the fungus. Cultures of K. pfeilii were also screened for enzyme production including amylase, protease, cellulose, and laccase. Evaluation of the potential of K. pfeilii mycelia to produce these industrially and economically important enzymes demonstrated both amylase and protease activity. However, for laccase and cellulose, no activity was detected. The second part of the present study aimed at optimizing biomass production by K. pfeilii in liquid culture media. FF Microplate containing 95 discreet carbon sources were employed to test for substrate utilization. Blanked readings above 0.1 were regarded as positive for utilization, and 4 substrates were selected as potential substrates and were included in liquid media. Media was evaluated for mycelial biomass production. Of the carbon sources tested sucrose proved to be the most suitable for supporting mycelial growth. The third part of the current study included investigating the diversity of microbial communities colonizing the rhizosheath of Stipagrostis ciliata var. capensis (the host plant of K. pfeilii) and these were identified by means of next-generation sequencing using Illumina Miseq. Bioinformatics tools were utilized in analyzing the data. Actinobacteria were found to be the most dominant bacterial phylum, followed by unclassified bacteria, Proteobacteria, and Acidobacteria. The top 25 sequences were selected and clustered into bacterial OTUs (at 97% threshold) which were assigned into 1 phylum (Actinobacteria), 1 family (Geodermatophilaceae) and 23 genera. This phylum is well known for its secondary metabolites. Streptomyces sp. was the most frequently encountered genus. The results from this study necessitate further investigations with regards to the function and evolution of fungal-bacterial associations. Wheather these bacteria have a contribution towards the truffle development, it is still not confirmed.
- Full Text:
- Authors: Krele, Viwe
- Date: 2019
- Subjects: Kalaharituber pfeilii , Pezizales -- South Africa , Desert plants -- South Africa , Truffle culture -- South Africa , Plant biochemical genetics , Enzymes -- Analysis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72257 , vital:30022
- Description: Dessert truffles are seasonal macro fungi and have been identified in several parts of the world including South Africa. The first part of the present study dealt with the assessment of the biologically active compounds of the Kalahari truffles found in the Northern Cape of South Africa. Truffles extracts (methanol, ethanol, aqueous) were investigated for their antimicrobial properties towards Gram-positive and Gram-negative bacteria. The results demonstrated that the truffle extracts tested had no inhibitory effects against the bacterial isolates. The truffle mycelial growth was also noted to be ineffective against the selected bacteria. The bacteria tested in the present study showed some antagonistic effects against the fungus. Cultures of K. pfeilii were also screened for enzyme production including amylase, protease, cellulose, and laccase. Evaluation of the potential of K. pfeilii mycelia to produce these industrially and economically important enzymes demonstrated both amylase and protease activity. However, for laccase and cellulose, no activity was detected. The second part of the present study aimed at optimizing biomass production by K. pfeilii in liquid culture media. FF Microplate containing 95 discreet carbon sources were employed to test for substrate utilization. Blanked readings above 0.1 were regarded as positive for utilization, and 4 substrates were selected as potential substrates and were included in liquid media. Media was evaluated for mycelial biomass production. Of the carbon sources tested sucrose proved to be the most suitable for supporting mycelial growth. The third part of the current study included investigating the diversity of microbial communities colonizing the rhizosheath of Stipagrostis ciliata var. capensis (the host plant of K. pfeilii) and these were identified by means of next-generation sequencing using Illumina Miseq. Bioinformatics tools were utilized in analyzing the data. Actinobacteria were found to be the most dominant bacterial phylum, followed by unclassified bacteria, Proteobacteria, and Acidobacteria. The top 25 sequences were selected and clustered into bacterial OTUs (at 97% threshold) which were assigned into 1 phylum (Actinobacteria), 1 family (Geodermatophilaceae) and 23 genera. This phylum is well known for its secondary metabolites. Streptomyces sp. was the most frequently encountered genus. The results from this study necessitate further investigations with regards to the function and evolution of fungal-bacterial associations. Wheather these bacteria have a contribution towards the truffle development, it is still not confirmed.
- Full Text:
Investigating soil microbial interactions of Portulacaria afra
- Authors: Fulmaka, Aviwe
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54598 , vital:26592
- Description: Portulacaria afra commonly known as Spekboom contributes significantly to carbon sequestration and has been widely planted in degraded areas of the Eastern Cape. Approximately 50% of planted cuttings do not survive although the cause of this decline is unknown. Like many indigenous plants, Spekboom forms a symbiotic relationship with mycorrhizal fungi and the interaction with rhizobacteria may enhance and improve plant growth and establishment. This study aims to investigate these relationships which will include a survey of the arbuscular mycorrhizal (AM) fungal populations associated with Spekboom, determination of the causal agent of Spekboom decline, isolation and identification of the associated rhizobacteria and investigation of their plant growth promotion properties and assessing the ability of arbuscular mycorrhizal fungi and selected rhizobacteria to enhance establishment and growth of Spekboom. Soil and root samples from selected trial sites were used to assess AM fungal spore abundance and colonisation; isolation, characterization, and identification of rhizobacteria and determine the interaction of the microbes on Spekboom growth and tolerance to Fusarium. AM spore abundance and percentage root colonisation did not differ between the three Spekboom plots. Molecular analyses of the SSU region from the plots showed 4 families of AM fungi and were identified as Ambisporaceae, Glomeraceae, Claroideoglomeraceae and Paraglomeraceae. A suspected Fusarium pathogen was isolated and molecularly identified. Pathogenicity tests indicated reduced Spekboom growth with poor root development. Thirty four rhizobacterial isolates were tested for various plant growth promoting abilities. Of these, 6 were able to produce IAA which may promote plant root growth, 27 siderophores and 23 were phosphate solubilisers. Bacterial isolates were molecularly identified to be from various species of Bacillus, with some Arthrobacter, Enterobacter, Pseudomonas and Microbacterium. Inoculation of Spekboom cuttings with mycorrhizal fungi and selected rhizobacterial isolates significantly improved shoot height. Spekboom cuttings challenged with Fusarium and inoculated with mycorrhizal fungi and two rhizobacterial isolates significantly improved growth. The inoculation of cuttings in the nursery with mycorrhizal fungi and selected rhizobacteria is recommended prior to establishing Spekboom in the field.
- Full Text:
- Authors: Fulmaka, Aviwe
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54598 , vital:26592
- Description: Portulacaria afra commonly known as Spekboom contributes significantly to carbon sequestration and has been widely planted in degraded areas of the Eastern Cape. Approximately 50% of planted cuttings do not survive although the cause of this decline is unknown. Like many indigenous plants, Spekboom forms a symbiotic relationship with mycorrhizal fungi and the interaction with rhizobacteria may enhance and improve plant growth and establishment. This study aims to investigate these relationships which will include a survey of the arbuscular mycorrhizal (AM) fungal populations associated with Spekboom, determination of the causal agent of Spekboom decline, isolation and identification of the associated rhizobacteria and investigation of their plant growth promotion properties and assessing the ability of arbuscular mycorrhizal fungi and selected rhizobacteria to enhance establishment and growth of Spekboom. Soil and root samples from selected trial sites were used to assess AM fungal spore abundance and colonisation; isolation, characterization, and identification of rhizobacteria and determine the interaction of the microbes on Spekboom growth and tolerance to Fusarium. AM spore abundance and percentage root colonisation did not differ between the three Spekboom plots. Molecular analyses of the SSU region from the plots showed 4 families of AM fungi and were identified as Ambisporaceae, Glomeraceae, Claroideoglomeraceae and Paraglomeraceae. A suspected Fusarium pathogen was isolated and molecularly identified. Pathogenicity tests indicated reduced Spekboom growth with poor root development. Thirty four rhizobacterial isolates were tested for various plant growth promoting abilities. Of these, 6 were able to produce IAA which may promote plant root growth, 27 siderophores and 23 were phosphate solubilisers. Bacterial isolates were molecularly identified to be from various species of Bacillus, with some Arthrobacter, Enterobacter, Pseudomonas and Microbacterium. Inoculation of Spekboom cuttings with mycorrhizal fungi and selected rhizobacterial isolates significantly improved shoot height. Spekboom cuttings challenged with Fusarium and inoculated with mycorrhizal fungi and two rhizobacterial isolates significantly improved growth. The inoculation of cuttings in the nursery with mycorrhizal fungi and selected rhizobacteria is recommended prior to establishing Spekboom in the field.
- Full Text:
Investigating the use of Arbuscular Mycorrhizas and Plant Growth Promoting Bacteria to improve the drought tolerance of maize (Zea mays L.)
- Authors: Moore, Nicolle Maureen
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54587 , vital:26591
- Description: Maize (Zea mays L.) is a direct staple food crop in Africa and remains an essential component of global food security, with maize crops accounting for over 60% of the total harvested area of annual food crops. Stress caused by drought and high soil salinity limits crop growth and productivity more than any other single environmental factor, with grain yield reductions up to 76% depending on the severity of the drought and the plant growth stage. Arbuscular mycorrhizal (AM) fungi and Plant Growth Promotion Rhizobacteria (PGPR) have previously been shown to improve tolerance of plants to drought stress through a number of chemical and physiological processes. The aim of this investigation was to determine whether mycorrhizal fungi and rhizobacteria adapted to drought and saline conditions and possessing plant growth promoting (PGP) traits were able to stimulate plant growth responses when applied to Zea mays seeds growing under greenhouse conditions Bacterial isolates selected were tolerant to concentrations of NaCl up to 600 mM and maintained 50% growth at low water potentials (-1.44 MPa). They were positive for Indole Acetic Acid (IAA) production, phosphate solubilisation and secretion of siderophores. Bacterial isolates showing plant growth promoting potential were identified using 16S rDNA gene sequencing as Achromobacter xylosoxidans strains A8 and C54 and Klebsiella oxytoca strain M1. Mixed inoculum was prepared from indigenous communities of mycorrhizas in soils sampled from the Cerebos Salt Pan and the Kalahari Desert. Mycorrhizal diversity was investigated using 454-Pyrosequencing which revealed that the community composition was dominated by species in the Ambispora, Glomus and Paraglomus genera with a rare component represented by species in the Redeckera, Archaeospora and Geosiphon genera. Microscopic examination of plant roots at the end of the trial revealed the presence of diagnostic mycorrhizal structures within the root cells, confirming that colonization was successful. Plant growth response to microbial inoculation was assessed by monitoring changes in plant photosynthetic capacity over the duration of a 7 week pot trial. A significant difference in photosynthetic and biomass data was observed between drought and well-watered groups but no mycorrhizal or bacterial treatment effect was evident within the groups, despite the high levels of colonization by mycorrhizas. These results suggest that the beneficial effects of mycorrhizal colonization may be primarily attributed to improved nutrient and mineral uptake in conditions where nutrients are limiting, resulting in improved growth. The improved growth may then have secondary effects on the plant‟s ability to withstand drought. Having controlled for nutrient deficiency, it was not evident in this study that mycorrhizal fungi were able to stimulate a change in plant physiology and confer drought tolerance under the conditions imposed.
- Full Text:
- Authors: Moore, Nicolle Maureen
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54587 , vital:26591
- Description: Maize (Zea mays L.) is a direct staple food crop in Africa and remains an essential component of global food security, with maize crops accounting for over 60% of the total harvested area of annual food crops. Stress caused by drought and high soil salinity limits crop growth and productivity more than any other single environmental factor, with grain yield reductions up to 76% depending on the severity of the drought and the plant growth stage. Arbuscular mycorrhizal (AM) fungi and Plant Growth Promotion Rhizobacteria (PGPR) have previously been shown to improve tolerance of plants to drought stress through a number of chemical and physiological processes. The aim of this investigation was to determine whether mycorrhizal fungi and rhizobacteria adapted to drought and saline conditions and possessing plant growth promoting (PGP) traits were able to stimulate plant growth responses when applied to Zea mays seeds growing under greenhouse conditions Bacterial isolates selected were tolerant to concentrations of NaCl up to 600 mM and maintained 50% growth at low water potentials (-1.44 MPa). They were positive for Indole Acetic Acid (IAA) production, phosphate solubilisation and secretion of siderophores. Bacterial isolates showing plant growth promoting potential were identified using 16S rDNA gene sequencing as Achromobacter xylosoxidans strains A8 and C54 and Klebsiella oxytoca strain M1. Mixed inoculum was prepared from indigenous communities of mycorrhizas in soils sampled from the Cerebos Salt Pan and the Kalahari Desert. Mycorrhizal diversity was investigated using 454-Pyrosequencing which revealed that the community composition was dominated by species in the Ambispora, Glomus and Paraglomus genera with a rare component represented by species in the Redeckera, Archaeospora and Geosiphon genera. Microscopic examination of plant roots at the end of the trial revealed the presence of diagnostic mycorrhizal structures within the root cells, confirming that colonization was successful. Plant growth response to microbial inoculation was assessed by monitoring changes in plant photosynthetic capacity over the duration of a 7 week pot trial. A significant difference in photosynthetic and biomass data was observed between drought and well-watered groups but no mycorrhizal or bacterial treatment effect was evident within the groups, despite the high levels of colonization by mycorrhizas. These results suggest that the beneficial effects of mycorrhizal colonization may be primarily attributed to improved nutrient and mineral uptake in conditions where nutrients are limiting, resulting in improved growth. The improved growth may then have secondary effects on the plant‟s ability to withstand drought. Having controlled for nutrient deficiency, it was not evident in this study that mycorrhizal fungi were able to stimulate a change in plant physiology and confer drought tolerance under the conditions imposed.
- Full Text:
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