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:
- Date Issued: 2019
- 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:
- Date Issued: 2019
Bioprospecting for amylases, cellulases and xylanases from ericoid associated fungi, their production and characterisation for the bio-economy
- Authors: Adeoyo, Olusegun Richard
- Date: 2018
- Subjects: Mycorrhizal fungi , Hydrolases , Ericaceae South Africa , Ericaceae Molecular aspects
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/64327 , vital:28533
- Description: South Africa is one of the most productive areas for ericaceous plants with about 850 identified species in the Cape Floral Region. The Albany Centre of Endemism where all fungi used in this study were isolated from, falls within this region. Ericaceous plants interact with some fungi via an association called the ericoid mycorrhizal (ERM) association. All fungi used in this study were isolated from roots of six ericaceous plants; Erica cerinthoides, Erica demissa, Erica chamissonis, Erica glumiflora, Erica caffra and Erica nemorosa. Fungal enzymes are known to play a significant role in the food, brewing, detergent, pharmaceutical and biofuel industries. The enzyme industry is among the major sectors of the world, and additional novel sources are being explored from time to time. This study focussed on amylases (amyloglucosidase, AMG), cellulases (endoglucanase) and xylanases (endo-1,4-P-xylanase) production from ERM fungal isolates. Out of the fifty-one (51), fungal isolates screened, ChemRU330 (Leohumicola sp.), EdRU083 and EdRU002 were among the fungi that had the highest activities of all the enzymes. They were tested for the ability to produce amylases and cellulases under different pH and nutritional conditions that included: carbon sources, nitrogen sources and metal ions, at an optimum temperature of 28°C in a modified Melin-Norkrans (MMN) liquid medium. Cellulase specific activity of 3.99, 2.18 and 4.31 (U/mg protein) for isolates EdRU083, EdRU002 and ChemRU330, respectively, was produced at an optimal pH of 5.0. For amylase, ChemRU330 had the highest specific activity of 1.11 U/mg protein while EdRU083 and EdRU02 had a specific activity of 0.80 and 0.92 U/mg protein, respectively, at the same pH with corresponding biomass yield of 113, 125 and 97 mg/50 ml, respectively. Increased enzyme activities and improved mycelial biomass production were obtained in the presence of supplements such as potassium, sodium, glucose, maltose, cellobiose, tryptone and peptone, while NaFe-EDTA and cobalt inhibited enzyme activity. ChemRU330 was selected to determine the consistency and amount of amylase, cellulase and xylanase formed after several in vitro subculturing events. AMG and endo-1,4-P-xylanase were found to have the most consistent production throughout the study period. The AMG was stable at 45oC (pH 5.0), retaining approximately 65% activity over a period of 24 h. The molecular mass of AMG and endo-1,4-P-xylanase were estimated to be 101 kDa and 72 kDa, respectively. The Km and kcat were 0.38 mg/ml and 70 s-1, respectively, using soluble starch (AMG). For endo-1,4-P-xylanase, the Km and Vmax were 0.93 mg/ml and 8.54 U/ml, respectively, using beechwood xylan (endo-1,4-P-xylanase) as substrate. Additionally, crude extracts of five root endophytes with unique morphological characteristics were screened for antibacterial properties and was followed by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). L. incrustata (ChemRU330) and Chaetomium sp. extracts exhibited varying degrees of inhibition against two Gram-positive and Gram-negative bacteria. The crude extract of L. incrustata was the most effective which was found to inhibit Staphylococcus aureus (MIC: 1 mg/ml), Bacillus subtilis (MIC: 2 mg/ml) and Proteus vulgaris (MIC: 16 mg/ml). The L. incrustata displayed potential for antibacterial production and could be considered as an additional source of new antimicrobial agents in drug and food preservation. Also, the three isolates used for enzyme production were identified to genus and species levels, i.e., Leohumicola incrustata (ChemRU330), Leohumicola sp. (EdRU083) and Oidiodendron sp. (EdRU002) using both ITS and Cox1 DNA regions. The molecular analysis results indicated that these ERM mycorrhizal fungi were similar to those successfully described by some researchers in South Africa and Australia. Therefore, this study opens new opportunities for exploring ERM fungal biomolecules for the bio-economy. The promising physicochemical properties, starch and xylan hydrolysis end- products, and being non-pathogenic make AMG and endo-1,4-P-xylanase potential candidates for future applications as additives in the food industry for the production of glucose, glucose syrups, high-fructose corn syrups, and as well as the production of bioethanol. Finally, the findings of this study revealed that it is possible to produce hydrolytic enzymes from ERM fungi in vitro using chemically defined media. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
- Authors: Adeoyo, Olusegun Richard
- Date: 2018
- Subjects: Mycorrhizal fungi , Hydrolases , Ericaceae South Africa , Ericaceae Molecular aspects
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/64327 , vital:28533
- Description: South Africa is one of the most productive areas for ericaceous plants with about 850 identified species in the Cape Floral Region. The Albany Centre of Endemism where all fungi used in this study were isolated from, falls within this region. Ericaceous plants interact with some fungi via an association called the ericoid mycorrhizal (ERM) association. All fungi used in this study were isolated from roots of six ericaceous plants; Erica cerinthoides, Erica demissa, Erica chamissonis, Erica glumiflora, Erica caffra and Erica nemorosa. Fungal enzymes are known to play a significant role in the food, brewing, detergent, pharmaceutical and biofuel industries. The enzyme industry is among the major sectors of the world, and additional novel sources are being explored from time to time. This study focussed on amylases (amyloglucosidase, AMG), cellulases (endoglucanase) and xylanases (endo-1,4-P-xylanase) production from ERM fungal isolates. Out of the fifty-one (51), fungal isolates screened, ChemRU330 (Leohumicola sp.), EdRU083 and EdRU002 were among the fungi that had the highest activities of all the enzymes. They were tested for the ability to produce amylases and cellulases under different pH and nutritional conditions that included: carbon sources, nitrogen sources and metal ions, at an optimum temperature of 28°C in a modified Melin-Norkrans (MMN) liquid medium. Cellulase specific activity of 3.99, 2.18 and 4.31 (U/mg protein) for isolates EdRU083, EdRU002 and ChemRU330, respectively, was produced at an optimal pH of 5.0. For amylase, ChemRU330 had the highest specific activity of 1.11 U/mg protein while EdRU083 and EdRU02 had a specific activity of 0.80 and 0.92 U/mg protein, respectively, at the same pH with corresponding biomass yield of 113, 125 and 97 mg/50 ml, respectively. Increased enzyme activities and improved mycelial biomass production were obtained in the presence of supplements such as potassium, sodium, glucose, maltose, cellobiose, tryptone and peptone, while NaFe-EDTA and cobalt inhibited enzyme activity. ChemRU330 was selected to determine the consistency and amount of amylase, cellulase and xylanase formed after several in vitro subculturing events. AMG and endo-1,4-P-xylanase were found to have the most consistent production throughout the study period. The AMG was stable at 45oC (pH 5.0), retaining approximately 65% activity over a period of 24 h. The molecular mass of AMG and endo-1,4-P-xylanase were estimated to be 101 kDa and 72 kDa, respectively. The Km and kcat were 0.38 mg/ml and 70 s-1, respectively, using soluble starch (AMG). For endo-1,4-P-xylanase, the Km and Vmax were 0.93 mg/ml and 8.54 U/ml, respectively, using beechwood xylan (endo-1,4-P-xylanase) as substrate. Additionally, crude extracts of five root endophytes with unique morphological characteristics were screened for antibacterial properties and was followed by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). L. incrustata (ChemRU330) and Chaetomium sp. extracts exhibited varying degrees of inhibition against two Gram-positive and Gram-negative bacteria. The crude extract of L. incrustata was the most effective which was found to inhibit Staphylococcus aureus (MIC: 1 mg/ml), Bacillus subtilis (MIC: 2 mg/ml) and Proteus vulgaris (MIC: 16 mg/ml). The L. incrustata displayed potential for antibacterial production and could be considered as an additional source of new antimicrobial agents in drug and food preservation. Also, the three isolates used for enzyme production were identified to genus and species levels, i.e., Leohumicola incrustata (ChemRU330), Leohumicola sp. (EdRU083) and Oidiodendron sp. (EdRU002) using both ITS and Cox1 DNA regions. The molecular analysis results indicated that these ERM mycorrhizal fungi were similar to those successfully described by some researchers in South Africa and Australia. Therefore, this study opens new opportunities for exploring ERM fungal biomolecules for the bio-economy. The promising physicochemical properties, starch and xylan hydrolysis end- products, and being non-pathogenic make AMG and endo-1,4-P-xylanase potential candidates for future applications as additives in the food industry for the production of glucose, glucose syrups, high-fructose corn syrups, and as well as the production of bioethanol. Finally, the findings of this study revealed that it is possible to produce hydrolytic enzymes from ERM fungi in vitro using chemically defined media. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
Production, purification, and characterisation of proteases from an ericoid mycorrhizal fungus, Oidiodendron maius
- Authors: Manyumwa, Colleen Varaidzo
- Date: 2018
- Subjects: Ascomycetes , Mycorrhizal fungi , Ericaceae , Proteolytic enzymes , Silver Recycling
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/62833 , vital:28298
- Description: The aim of this study was to produce, purify and characterise proteases from the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b/KP119480), as well as to explore their potential application in the recovery of silver from X-ray film. Firstly, the growth of the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b), was studied, and its ability to produce proteolytic enzymes was investigated. O. maius proved to grow well in the dark, submerged in Modified Melin Norkran’s liquid medium at a pH of 5 and at 25°C. Pure cultures of the fungus were maintained on Potato Dextrose Agar (PDA). The fungus grew on PDA plates containing different substrates including haemoglobin, casein, gelatin as well as azocasein. Zones of clearance, however, were only observed on plates containing gelatin after treatment with mercuric chloride, HgCl2. Proteases were successfully produced after 14 days when gelatin was incorporated into the growth medium. After production of the proteases, purification and characterisation of the enzymes was performed. Purification of the enzymes was performed by acetone precipitation followed by ultrafiltration with 50 kDa and 30 kDa cut off membrane filters. A final purification fold of approximately 37.6 was achieved. Unusual yields of above 100% were observed after each purification step with the final yield achieved being 196% with a final specific activity of 2707 U/mg. SDS-PAGE revealed a protease band of 35 kDa which was also visible on the zymogram at approximately 36 kDa. The zymogram showed clear hydrolysis bands against a blue background after staining with Coomassie Brilliant Blue. Physico-chemical characterisation of the protease revealed its pH optimum to be pH 3.0 and its temperature optimum 68°C. Another peak was observed on the pH profile at pH 7.0. The protease exhibited high thermostability at temperatures 37°C, 80°C as well as 100°C with the enzyme retaining close to 50% of its initial activity after 4 h of exposure to all three temperatures. All ions tested for their effects on the proteases, except Ca2+, enhanced protease activity. Ca2+ did not exhibit any significant effect on the enzyme’s activity while Zn2+ had the highest effect, enhancing enzyme activity by 305%. The proteases, however, were not significantly inhibited by EDTA, a metal chelating agent and a known metalloprotease inhibitor. The enzyme was classified as an aspartic protease due to complete inhibition by 25 μM of pepstatin A, coupled to its low pH optimum of 3.0. Addition of trans-Epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64), a cysteine protease inhibitor, and 2-mercaptoethanol increased protease activity. The proteases exhibited a narrow substrate specificity towards gelatin and no other substrate. Substrate kinetics values were plotted on a Michaelis-Menten Graph and showed that the enzyme had a Vmax of 55.25 U/ml and a Km of 2.7 mg/ml gelatin. A low Km indicated that the protease had a high affinity for gelatin. Silver recovery studies from X-ray film revealed the proteases’ capability to remove silver from X-ray film, leaving the film intact. The recovery of silver was perceived visually, by film observation, as well as by scan electron microscopy (SEM) images, where clearance of the film was observed after incubation with the enzyme. Energy dispersive X-ray spectroscopy (EDS) profiles also confirmed removal of silver from the film, with a Ag peak showing on the profile of the film before treatment with the proteases and no peak after treatment. The crude protease sample was, however, catalytically more efficient compared to the partially purified sample. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
- Authors: Manyumwa, Colleen Varaidzo
- Date: 2018
- Subjects: Ascomycetes , Mycorrhizal fungi , Ericaceae , Proteolytic enzymes , Silver Recycling
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/62833 , vital:28298
- Description: The aim of this study was to produce, purify and characterise proteases from the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b/KP119480), as well as to explore their potential application in the recovery of silver from X-ray film. Firstly, the growth of the ericoid mycorrhizal fungus, Oidiodendron maius (CafRU082b), was studied, and its ability to produce proteolytic enzymes was investigated. O. maius proved to grow well in the dark, submerged in Modified Melin Norkran’s liquid medium at a pH of 5 and at 25°C. Pure cultures of the fungus were maintained on Potato Dextrose Agar (PDA). The fungus grew on PDA plates containing different substrates including haemoglobin, casein, gelatin as well as azocasein. Zones of clearance, however, were only observed on plates containing gelatin after treatment with mercuric chloride, HgCl2. Proteases were successfully produced after 14 days when gelatin was incorporated into the growth medium. After production of the proteases, purification and characterisation of the enzymes was performed. Purification of the enzymes was performed by acetone precipitation followed by ultrafiltration with 50 kDa and 30 kDa cut off membrane filters. A final purification fold of approximately 37.6 was achieved. Unusual yields of above 100% were observed after each purification step with the final yield achieved being 196% with a final specific activity of 2707 U/mg. SDS-PAGE revealed a protease band of 35 kDa which was also visible on the zymogram at approximately 36 kDa. The zymogram showed clear hydrolysis bands against a blue background after staining with Coomassie Brilliant Blue. Physico-chemical characterisation of the protease revealed its pH optimum to be pH 3.0 and its temperature optimum 68°C. Another peak was observed on the pH profile at pH 7.0. The protease exhibited high thermostability at temperatures 37°C, 80°C as well as 100°C with the enzyme retaining close to 50% of its initial activity after 4 h of exposure to all three temperatures. All ions tested for their effects on the proteases, except Ca2+, enhanced protease activity. Ca2+ did not exhibit any significant effect on the enzyme’s activity while Zn2+ had the highest effect, enhancing enzyme activity by 305%. The proteases, however, were not significantly inhibited by EDTA, a metal chelating agent and a known metalloprotease inhibitor. The enzyme was classified as an aspartic protease due to complete inhibition by 25 μM of pepstatin A, coupled to its low pH optimum of 3.0. Addition of trans-Epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64), a cysteine protease inhibitor, and 2-mercaptoethanol increased protease activity. The proteases exhibited a narrow substrate specificity towards gelatin and no other substrate. Substrate kinetics values were plotted on a Michaelis-Menten Graph and showed that the enzyme had a Vmax of 55.25 U/ml and a Km of 2.7 mg/ml gelatin. A low Km indicated that the protease had a high affinity for gelatin. Silver recovery studies from X-ray film revealed the proteases’ capability to remove silver from X-ray film, leaving the film intact. The recovery of silver was perceived visually, by film observation, as well as by scan electron microscopy (SEM) images, where clearance of the film was observed after incubation with the enzyme. Energy dispersive X-ray spectroscopy (EDS) profiles also confirmed removal of silver from the film, with a Ag peak showing on the profile of the film before treatment with the proteases and no peak after treatment. The crude protease sample was, however, catalytically more efficient compared to the partially purified sample. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
The diversity of root fungi associated with Erica species occurring in the Albany Centre of Endemism
- Authors: Bizabani, Christine
- Date: 2015
- Subjects: Ericaceae , Ericas , Roots (Botany) -- Diseases and pests , Mycorrhizal fungi , Polymerase chain reaction , Fungi -- Classification
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4160 , http://hdl.handle.net/10962/d1018575
- Description: South Africa has the highest species diversity of ericaceous plants belonging to the Erica genus. There are over 850 identified species in the Cape Floral Region. The Albany Centre of Endemism (ACOE) is located within this region and is a hotspot of diversity consisting of various plant genera. The success of Erica plants is ubiquitously attributed to mycorrhizal relationships they engage in with a diverse group of fungi. This symbiosis is known as the ericoid mycorrhizal (ERM) association. The overall aim of this study was to establish the diversity of root fungi associated with Erica plants using morphological, molecular and 454 pyrosequencing techniques. Six Erica species were identified using leaf and flower morphology according to taxonomic keys. The identified plants were Erica cerinthoides, Erica demissa, Erica chamissonis, Erica glumiflora, Erica caffra and Erica nemorosa. Roots from sampled plants were stained and examined microscopically to determine their mycorrhizal status. Ericoid mycorrhizal associations together with dark septate endophyte (DSE) structures and hyphae that did not form any specific structure were observed in all the roots. In addition arbuscular mycorrhizal (AM) structures in the form of vesicles were detected in E. glumiflora and E. cerinthoides. In order to identify the culturable fungi associated with the respective hosts, sterilised roots were placed on various culture media for cultivation. Thereafter isolated fungi were morphologically classified into 67 morphotypes. These were mostly sterile and darkly pigmented. Non-sporulating mycelia of variable colouration such as white, cream-yellowish, beige, green and brown were also observed. Further identification was carried out using molecular techniques. DNA was extracted separately from pure cultures and amplified using ITS1 and ITS4 primers in a polymerase chain reaction (PCR). Thereafter sequencing and Basic Local Alignment Search Tool (BLAST) were used to identify the isolates to generic level. The fungi were taxonomically classified into 54 operational taxonomic units and 94 percent were Ascomycetes and Helotiales was the dominant order. Unclassified Helotiales with affinities to fungi currently identified as Epacrid root fungus was common in all hosts. Other isolates that were identified included Oidiodendron, Meliniomyces, Phialocephala, Cadophora, Lachnum, Leohumicola Cryptosporiopsis, Chaetomium, Acremonium and Epicoccum species. Basidiomycetes were represented by two OTUs belonging to the genus Mycena. Four OTUs comprised fungi that had no significant alignments in the reference databases. Direct root DNA extraction together with 454 pyrosequencing was used to detect the diversity of culturable and unculturable fungi associated with the identified hosts. The ITS2 region was targeted for sequencing. Although Ascomycetes remained the dominant phyla, Basidiomycetes were also detected in all host plants. Glomeromycota was present in E. caffra and E. cerinthoides. Helotiales was dominant in all Erica plants with the exception of E. cerinthoides and E. chamissonis which were dominated by the order Chaetothyriales. The OTUs identified to genus level included Epacris pulchella root fungus, Oidiodendron cf. maius, Acremonium implicatum, Leohumicola, Lachnum, Capronia and Mycena species. Culture-based techniques and pyrosequencing detected similar fungal composition comprising Ascomycetes, while, pyrosequencing was able to detect Glomeromycetes and Basidiomycetes.
- Full Text:
- Date Issued: 2015
The diversity of root fungi associated with Erica species occurring in the Albany Centre of Endemism
- Authors: Bizabani, Christine
- Date: 2015
- Subjects: Ericaceae , Ericas , Roots (Botany) -- Diseases and pests , Mycorrhizal fungi , Polymerase chain reaction , Fungi -- Classification
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4160 , http://hdl.handle.net/10962/d1018575
- Description: South Africa has the highest species diversity of ericaceous plants belonging to the Erica genus. There are over 850 identified species in the Cape Floral Region. The Albany Centre of Endemism (ACOE) is located within this region and is a hotspot of diversity consisting of various plant genera. The success of Erica plants is ubiquitously attributed to mycorrhizal relationships they engage in with a diverse group of fungi. This symbiosis is known as the ericoid mycorrhizal (ERM) association. The overall aim of this study was to establish the diversity of root fungi associated with Erica plants using morphological, molecular and 454 pyrosequencing techniques. Six Erica species were identified using leaf and flower morphology according to taxonomic keys. The identified plants were Erica cerinthoides, Erica demissa, Erica chamissonis, Erica glumiflora, Erica caffra and Erica nemorosa. Roots from sampled plants were stained and examined microscopically to determine their mycorrhizal status. Ericoid mycorrhizal associations together with dark septate endophyte (DSE) structures and hyphae that did not form any specific structure were observed in all the roots. In addition arbuscular mycorrhizal (AM) structures in the form of vesicles were detected in E. glumiflora and E. cerinthoides. In order to identify the culturable fungi associated with the respective hosts, sterilised roots were placed on various culture media for cultivation. Thereafter isolated fungi were morphologically classified into 67 morphotypes. These were mostly sterile and darkly pigmented. Non-sporulating mycelia of variable colouration such as white, cream-yellowish, beige, green and brown were also observed. Further identification was carried out using molecular techniques. DNA was extracted separately from pure cultures and amplified using ITS1 and ITS4 primers in a polymerase chain reaction (PCR). Thereafter sequencing and Basic Local Alignment Search Tool (BLAST) were used to identify the isolates to generic level. The fungi were taxonomically classified into 54 operational taxonomic units and 94 percent were Ascomycetes and Helotiales was the dominant order. Unclassified Helotiales with affinities to fungi currently identified as Epacrid root fungus was common in all hosts. Other isolates that were identified included Oidiodendron, Meliniomyces, Phialocephala, Cadophora, Lachnum, Leohumicola Cryptosporiopsis, Chaetomium, Acremonium and Epicoccum species. Basidiomycetes were represented by two OTUs belonging to the genus Mycena. Four OTUs comprised fungi that had no significant alignments in the reference databases. Direct root DNA extraction together with 454 pyrosequencing was used to detect the diversity of culturable and unculturable fungi associated with the identified hosts. The ITS2 region was targeted for sequencing. Although Ascomycetes remained the dominant phyla, Basidiomycetes were also detected in all host plants. Glomeromycota was present in E. caffra and E. cerinthoides. Helotiales was dominant in all Erica plants with the exception of E. cerinthoides and E. chamissonis which were dominated by the order Chaetothyriales. The OTUs identified to genus level included Epacris pulchella root fungus, Oidiodendron cf. maius, Acremonium implicatum, Leohumicola, Lachnum, Capronia and Mycena species. Culture-based techniques and pyrosequencing detected similar fungal composition comprising Ascomycetes, while, pyrosequencing was able to detect Glomeromycetes and Basidiomycetes.
- Full Text:
- Date Issued: 2015
Investigating the role of mycorrhizal fungi and associated bacteria in promoting growth of citrus seedlings
- Authors: Sitole, Phumeza
- Date: 2014
- Subjects: Mycorrhizal fungi , Citrus -- South Africa , Citrus -- Diseases and pests -- Biological control -- South Africa , Fungi as biological pest control agents , Bacteria , Phytophthora , Pythium , Indoleacetic acid
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4111 , http://hdl.handle.net/10962/d1013033
- Description: South Africa is the world's second largest exporter of fresh citrus and is ranked 14th in citrus production. Fungal pathogens such as Phytophthora and Pythium cause economic losses as a result of root rot and brown rot. Mycorrhizal fungi are specialized members of the fungal community forming a mutualistic relationship with plant roots. Mycorrhizal fungal structures are known to associate with other soil microorganisms and these may contribute to improved plant growth. A diverse group of bacteria that interact with the mycorrhizal fungi are known as Mycorrhizal Helper Bacteria (MHB). The aim of this study was to investigate the role of arbuscular mycorrhiza and associated bacteria isolated from spores and determine whether they had any plant growth promoting potential. A total of 19 bacteria were isolated from arbuscular mycorrhizal spores and were molecularly identified as belonging to several Bacillus, Micrococcus, Onchrobactrum and Staphylococcus sp. All bacterial isolates were tested for plant growth promotion abilities. One Bacillus isolate was able to solubilise phosphate. Four isolates Micrococcus sp, Micrococcus leteus, Ochrobacterum sp and Ochrobacterum antropi were able to produce Indole Acetic Acid and three isolates showed potential to reduce growth of Phytophthora nicotianae, P. citrocola and P. citrophthora in in vitro plate cultures. Further tests using culture supernatants of the Bacillus sp, Micrococcus sp and Bacillus cereus confirmed their ability to inhibit or reduce growth of the three Phytophthora species in a 96 well bioassay. Bacillus sp and Bacillus cereus were able to inhibit Phytophthora spp by 95 to 100 % and Micrococcus spp was able to decrease pathogen growth by 60 to 94 %. These bacterial isolates were further evaluated for plant growth promoting abilities on citrus rough lemon seedlings alone or in combination with arbuscular mycorrhizal inoculum. Bacterial and mycorrhizal inoculants influence the increase in shoot and root biomass. Bacillus cereus in combination with mycorrhizal inoculum significantly increased seedling shoot to root ratio while root biomass was significantly increased with mycorrhizal inoculation. Due to the short duration of the trial mycorrhizal colonisation could not be assessed. It is evident that selected combinations of bacteria and mycorrhizal fungi could promote citrus seedling growth and potentially improve seedling health. Further studies under nursery conditions are recommended.
- Full Text:
- Date Issued: 2014
- Authors: Sitole, Phumeza
- Date: 2014
- Subjects: Mycorrhizal fungi , Citrus -- South Africa , Citrus -- Diseases and pests -- Biological control -- South Africa , Fungi as biological pest control agents , Bacteria , Phytophthora , Pythium , Indoleacetic acid
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4111 , http://hdl.handle.net/10962/d1013033
- Description: South Africa is the world's second largest exporter of fresh citrus and is ranked 14th in citrus production. Fungal pathogens such as Phytophthora and Pythium cause economic losses as a result of root rot and brown rot. Mycorrhizal fungi are specialized members of the fungal community forming a mutualistic relationship with plant roots. Mycorrhizal fungal structures are known to associate with other soil microorganisms and these may contribute to improved plant growth. A diverse group of bacteria that interact with the mycorrhizal fungi are known as Mycorrhizal Helper Bacteria (MHB). The aim of this study was to investigate the role of arbuscular mycorrhiza and associated bacteria isolated from spores and determine whether they had any plant growth promoting potential. A total of 19 bacteria were isolated from arbuscular mycorrhizal spores and were molecularly identified as belonging to several Bacillus, Micrococcus, Onchrobactrum and Staphylococcus sp. All bacterial isolates were tested for plant growth promotion abilities. One Bacillus isolate was able to solubilise phosphate. Four isolates Micrococcus sp, Micrococcus leteus, Ochrobacterum sp and Ochrobacterum antropi were able to produce Indole Acetic Acid and three isolates showed potential to reduce growth of Phytophthora nicotianae, P. citrocola and P. citrophthora in in vitro plate cultures. Further tests using culture supernatants of the Bacillus sp, Micrococcus sp and Bacillus cereus confirmed their ability to inhibit or reduce growth of the three Phytophthora species in a 96 well bioassay. Bacillus sp and Bacillus cereus were able to inhibit Phytophthora spp by 95 to 100 % and Micrococcus spp was able to decrease pathogen growth by 60 to 94 %. These bacterial isolates were further evaluated for plant growth promoting abilities on citrus rough lemon seedlings alone or in combination with arbuscular mycorrhizal inoculum. Bacterial and mycorrhizal inoculants influence the increase in shoot and root biomass. Bacillus cereus in combination with mycorrhizal inoculum significantly increased seedling shoot to root ratio while root biomass was significantly increased with mycorrhizal inoculation. Due to the short duration of the trial mycorrhizal colonisation could not be assessed. It is evident that selected combinations of bacteria and mycorrhizal fungi could promote citrus seedling growth and potentially improve seedling health. Further studies under nursery conditions are recommended.
- Full Text:
- Date Issued: 2014
Interactions of arbuscular mycorrhizal fungi and spore-associated bacteria
- Authors: Ridsdale, Carmen Jane
- Date: 2013
- Subjects: Mycorrhizal fungi , Host plants , Bacteria
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4159 , http://hdl.handle.net/10962/d1018269
- Description: Arbuscular mycorrhizal (AM) fungi are naturally occurring in roots of terrestrial plants. AM fungi are capable of benefiting the host plant through various mechanisms such as enhanced nutrient supply, alleviation of environmental stress and inhibition of plant fungal pathogens. AM fungal spore-associated bacteria have been previously isolated and shown to have plant growthpromoting (PGP) abilities by several authors. Some bacterial isolates are able to promote AM fungal colonisation of host plants and are known to be mycorrhizal helper bacteria (MHB). This study focused on the isolation of AM fungal spore-associated bacteria, characterization of the isolates according to plant growth promoting abilities and evaluation of their potential to enhance plant growth and mycorrhizal colonisation. AM fungi were extracted from soils sampled from natural indigenous forest sources, raspberry (Rubus idaeus cv. Heritage) and strawberry (Fragaria ananassa) farms in South Africa and from a raspberry (Rubus idaeus cv. Autumn Bliss) plantation in Argentina. A total of 52 sporeassociated bacteria were isolated from the external and internal surfaces of AM fungal spore morphotypes from the two countries. The bacterial isolates were evaluated for their PGP abilities such as phosphate solubilisation, indole-3-acetic acid production, ammonia production and inhibition of the fungal pathogens Fusarium oxysporum and Phythophthora nicotianae through mechanisms such as siderophore and/ or hydrolytic enzyme production. A total of 23 bacterial isolates from both South Africa and Argentina showing the most potential to be PGP, were identified molecularly as belonging to the genera Acinetobacter, Alcaligenes, Bacillus, Microbacterium, Micrococcus, Serratia and Staphylococcus. The ability of ten selected bacterial isolates showing multiple PGP capacity were evaluated for their plant growth promotion and mycorrhizal colonisation enhancement ability on raspberry (Rubus idaeus cv. Meeker). Significant differences in increased shoot and root dry weights were shown by the treatments compared to the uninoculated control. The highest increase in shoot and root dry weights were shown by South African (Bacillus mycoides) and Argentinean (Alcaligenes faecalis) isolates. AM fungal colonisation was significantly enhanced by the South African (Bacillus mycoides) and Argentinean (Micrococcus luteus) isolates compared to the AM fungal singly inoculated control.
- Full Text:
- Date Issued: 2013
- Authors: Ridsdale, Carmen Jane
- Date: 2013
- Subjects: Mycorrhizal fungi , Host plants , Bacteria
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4159 , http://hdl.handle.net/10962/d1018269
- Description: Arbuscular mycorrhizal (AM) fungi are naturally occurring in roots of terrestrial plants. AM fungi are capable of benefiting the host plant through various mechanisms such as enhanced nutrient supply, alleviation of environmental stress and inhibition of plant fungal pathogens. AM fungal spore-associated bacteria have been previously isolated and shown to have plant growthpromoting (PGP) abilities by several authors. Some bacterial isolates are able to promote AM fungal colonisation of host plants and are known to be mycorrhizal helper bacteria (MHB). This study focused on the isolation of AM fungal spore-associated bacteria, characterization of the isolates according to plant growth promoting abilities and evaluation of their potential to enhance plant growth and mycorrhizal colonisation. AM fungi were extracted from soils sampled from natural indigenous forest sources, raspberry (Rubus idaeus cv. Heritage) and strawberry (Fragaria ananassa) farms in South Africa and from a raspberry (Rubus idaeus cv. Autumn Bliss) plantation in Argentina. A total of 52 sporeassociated bacteria were isolated from the external and internal surfaces of AM fungal spore morphotypes from the two countries. The bacterial isolates were evaluated for their PGP abilities such as phosphate solubilisation, indole-3-acetic acid production, ammonia production and inhibition of the fungal pathogens Fusarium oxysporum and Phythophthora nicotianae through mechanisms such as siderophore and/ or hydrolytic enzyme production. A total of 23 bacterial isolates from both South Africa and Argentina showing the most potential to be PGP, were identified molecularly as belonging to the genera Acinetobacter, Alcaligenes, Bacillus, Microbacterium, Micrococcus, Serratia and Staphylococcus. The ability of ten selected bacterial isolates showing multiple PGP capacity were evaluated for their plant growth promotion and mycorrhizal colonisation enhancement ability on raspberry (Rubus idaeus cv. Meeker). Significant differences in increased shoot and root dry weights were shown by the treatments compared to the uninoculated control. The highest increase in shoot and root dry weights were shown by South African (Bacillus mycoides) and Argentinean (Alcaligenes faecalis) isolates. AM fungal colonisation was significantly enhanced by the South African (Bacillus mycoides) and Argentinean (Micrococcus luteus) isolates compared to the AM fungal singly inoculated control.
- Full Text:
- Date Issued: 2013
Interaction between arbuscular mycorrhizal fungi and soil microbial populations in the rhizosphere
- Authors: Ike-Izundu, Nnenna Esther
- Date: 2008
- Subjects: Mycorrhizas , Mycorrhizal fungi , Vesicular-arbuscular mycorrhizas , Soil microbiology , Rhizosphere , Revegetation , Restoration ecology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3962 , http://hdl.handle.net/10962/d1004021 , Mycorrhizas , Mycorrhizal fungi , Vesicular-arbuscular mycorrhizas , Soil microbiology , Rhizosphere , Revegetation , Restoration ecology
- Description: This study examined the rehabilitation potential of AM fungi with organic and inorganic fertilisers under pot and field trial conditions as well as their interaction with rhizospheric organisms and specific functional groups. In addition, the study highlighted the effects of land-use management on AM fungal populations in soil and the mycorrhizal status of some selected plants from one of the study sites. The study focussed on two sites that differ in operational activities and these included a mined area that was to be rehabilitated and a commercial farming site. A pot trial was conducted using an overburdened soil resulting from kaolin clay mining. Pots were seeded with Cynodon dactylon and treated with either Organic Tea or NPK (3:1:5) fertiliser, with or without AM fungal inoculum. The compatibility of these fertilisers with AM fungi was assessed by plant growth and percentage root colonisation. Maximum shoot height and plant biomass were observed at the 28th week with NPK (3:1:5) fertiliser supporting mycorrhizal colonisation by 80%. The result indicated the potential of AM fungi to be used in rehabilitation with minimal phosphate fertiliser. Similarly, a field trial was set-up using 17 x 17 m[superscript 2] plots in the mining site that were treated with the same organic and inorganic fertilisers as well as with AM fungal inoculum in different combinations. The interaction between AM fungi and soil microbial population was determined using culture dependent and culture independent techniques. The culture dependent technique involved the use of soil dilution and plating on general purpose and selective media. The result showed that there was no change in the total culturable bacterial number in the untreated and AM fungal treated plots, while a change in species composition was observed in the functional groups. Different functional groups identified included nitrogen fixing bacteria, pseudomonads, actinomycetes, phosphate solubilisers and the fungal counterparts. Gram-positive bacteria were observed as the predominant phenotypic type, while nitrogen fixers and actinomycetes were the predominant functional groups. Species identified from each functional group were Pseudomonas fulva, Bacillus megaterium, Streptomyces and actinomycetales bacteria. Meanwhile, fungi such as Ampelomyces, Fusarium, Penicillium, Aspergillus, Cephalosporium and Exserohilium were identified morphologically and molecularly. Furthermore, the mining site had a significantly higher bacterial number than the farming site thereby indicating the effects of land-use management on culturable bacterial numbers. The culture independent technique was carried out by cloning of the bacterial 16S rDNA and sequencing. Identified clones were Bradyrhizobium, Propionibacterium and Sporichthya. A cladogram constructed with the nucleotides sequences of identified functional species, clones and closely related nucleotide sequences from the Genbank indicated that nucleotide sequences differed in terms of the method used. The activity and establishment of the introduced AM fungal population was determined by spore enumeration, infectivity assay, percentage root colonisation and assessment of glomalin concentrations. The results indicated that the two land use types affected AM fungal populations. However, the establishment of AM fungi in the farming site was more successful than in the mining site as indicated by the higher infectivity pontential. Selected host plants, which were collected around the mine area, were observed to be mainly colonised by AM fungi and these were identified as Pentzia incana, Elytropappus rhinocerotis, Euphorbia meloformis, Selago corymbosa, Albuca canadensis and Helichrysum rosum. These plant species were able to thrive under harsh environmental conditions, thereby indicating their potential use as rehabilitation host plants. Generally, the findings of this study has provided an insight into the interaction between arbuscular mycorrhizal fungi and other soil microorganisms in two fields with differing land use management practices.
- Full Text:
- Date Issued: 2008
- Authors: Ike-Izundu, Nnenna Esther
- Date: 2008
- Subjects: Mycorrhizas , Mycorrhizal fungi , Vesicular-arbuscular mycorrhizas , Soil microbiology , Rhizosphere , Revegetation , Restoration ecology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3962 , http://hdl.handle.net/10962/d1004021 , Mycorrhizas , Mycorrhizal fungi , Vesicular-arbuscular mycorrhizas , Soil microbiology , Rhizosphere , Revegetation , Restoration ecology
- Description: This study examined the rehabilitation potential of AM fungi with organic and inorganic fertilisers under pot and field trial conditions as well as their interaction with rhizospheric organisms and specific functional groups. In addition, the study highlighted the effects of land-use management on AM fungal populations in soil and the mycorrhizal status of some selected plants from one of the study sites. The study focussed on two sites that differ in operational activities and these included a mined area that was to be rehabilitated and a commercial farming site. A pot trial was conducted using an overburdened soil resulting from kaolin clay mining. Pots were seeded with Cynodon dactylon and treated with either Organic Tea or NPK (3:1:5) fertiliser, with or without AM fungal inoculum. The compatibility of these fertilisers with AM fungi was assessed by plant growth and percentage root colonisation. Maximum shoot height and plant biomass were observed at the 28th week with NPK (3:1:5) fertiliser supporting mycorrhizal colonisation by 80%. The result indicated the potential of AM fungi to be used in rehabilitation with minimal phosphate fertiliser. Similarly, a field trial was set-up using 17 x 17 m[superscript 2] plots in the mining site that were treated with the same organic and inorganic fertilisers as well as with AM fungal inoculum in different combinations. The interaction between AM fungi and soil microbial population was determined using culture dependent and culture independent techniques. The culture dependent technique involved the use of soil dilution and plating on general purpose and selective media. The result showed that there was no change in the total culturable bacterial number in the untreated and AM fungal treated plots, while a change in species composition was observed in the functional groups. Different functional groups identified included nitrogen fixing bacteria, pseudomonads, actinomycetes, phosphate solubilisers and the fungal counterparts. Gram-positive bacteria were observed as the predominant phenotypic type, while nitrogen fixers and actinomycetes were the predominant functional groups. Species identified from each functional group were Pseudomonas fulva, Bacillus megaterium, Streptomyces and actinomycetales bacteria. Meanwhile, fungi such as Ampelomyces, Fusarium, Penicillium, Aspergillus, Cephalosporium and Exserohilium were identified morphologically and molecularly. Furthermore, the mining site had a significantly higher bacterial number than the farming site thereby indicating the effects of land-use management on culturable bacterial numbers. The culture independent technique was carried out by cloning of the bacterial 16S rDNA and sequencing. Identified clones were Bradyrhizobium, Propionibacterium and Sporichthya. A cladogram constructed with the nucleotides sequences of identified functional species, clones and closely related nucleotide sequences from the Genbank indicated that nucleotide sequences differed in terms of the method used. The activity and establishment of the introduced AM fungal population was determined by spore enumeration, infectivity assay, percentage root colonisation and assessment of glomalin concentrations. The results indicated that the two land use types affected AM fungal populations. However, the establishment of AM fungi in the farming site was more successful than in the mining site as indicated by the higher infectivity pontential. Selected host plants, which were collected around the mine area, were observed to be mainly colonised by AM fungi and these were identified as Pentzia incana, Elytropappus rhinocerotis, Euphorbia meloformis, Selago corymbosa, Albuca canadensis and Helichrysum rosum. These plant species were able to thrive under harsh environmental conditions, thereby indicating their potential use as rehabilitation host plants. Generally, the findings of this study has provided an insight into the interaction between arbuscular mycorrhizal fungi and other soil microorganisms in two fields with differing land use management practices.
- Full Text:
- Date Issued: 2008
Investigating the effect of Glomus etunicatum colonization on structure and phloem transport in roots of Eragrostis curvula (Umgeni)
- Authors: Skinner, Amy
- Date: 2007
- Subjects: Glomus (Fungi) , Phloem , Plant translocation , Weeping lovegrass , Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Mycorrhizas
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4227 , http://hdl.handle.net/10962/d1003796 , Glomus (Fungi) , Phloem , Plant translocation , Weeping lovegrass , Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Mycorrhizas
- Description: The symbiotic unit of an arbuscular mycorrhizal fungus and its host is able toachieve and maintain far higher inflow of nutrients than non-mycorrhizal roots. The colonization strategy of the mycobiont within the plant is intrinsic to the symbiosis with respect to both structural adaptations and nutrient exchange. An investigation into the effect of Glomus etunicatum colonization on the structure and phloem transport in Eragrostis curvula (Umgeni) allowed for greater insight into the dynamic of the symbiosis. The combined use of stains (such as Trypan Blue, Chlorazol Black, Safranin and Fast Green), and techniques, (such as freeze-microtome transverse sectioning and permanent slide preparations) contributed to a successful general observation of an intermediate colonization strategy using light microscopy methods. However, clarity into structural detail of mycorrhizal forms required electron microscopy studies. The SEM method used with freeze fracture was a relatively quick and simple method allowing for the observation of surface and internal features. The TEM method allowed for highresolution images providing insight into the variations in the apoplasmic compartmental form, and how this may relate to the function of the symbiosis with regard to fungal coils or arbuscules. The apoplasmic nature of mycorrhizas was substantiated and no symplasmic connections were found between symbionts. Fluorescence studies demonstrated that 5,6-carboxyfluorescein was transported through the phloem into the roots of E. curvula, but remained predominantly in the root phloem. Unloading only occurred in optimal nutrient exchange areas of meristimatic lateral or apical growth regions. It was not possible, using fluorescence techniques and related equipment available, to conclusively establish if there were symplasmic connections between the mycobiont and its host or if bidirectional transfer of nutrients occurred at the same interface.
- Full Text:
- Date Issued: 2007
- Authors: Skinner, Amy
- Date: 2007
- Subjects: Glomus (Fungi) , Phloem , Plant translocation , Weeping lovegrass , Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Mycorrhizas
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4227 , http://hdl.handle.net/10962/d1003796 , Glomus (Fungi) , Phloem , Plant translocation , Weeping lovegrass , Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Mycorrhizas
- Description: The symbiotic unit of an arbuscular mycorrhizal fungus and its host is able toachieve and maintain far higher inflow of nutrients than non-mycorrhizal roots. The colonization strategy of the mycobiont within the plant is intrinsic to the symbiosis with respect to both structural adaptations and nutrient exchange. An investigation into the effect of Glomus etunicatum colonization on the structure and phloem transport in Eragrostis curvula (Umgeni) allowed for greater insight into the dynamic of the symbiosis. The combined use of stains (such as Trypan Blue, Chlorazol Black, Safranin and Fast Green), and techniques, (such as freeze-microtome transverse sectioning and permanent slide preparations) contributed to a successful general observation of an intermediate colonization strategy using light microscopy methods. However, clarity into structural detail of mycorrhizal forms required electron microscopy studies. The SEM method used with freeze fracture was a relatively quick and simple method allowing for the observation of surface and internal features. The TEM method allowed for highresolution images providing insight into the variations in the apoplasmic compartmental form, and how this may relate to the function of the symbiosis with regard to fungal coils or arbuscules. The apoplasmic nature of mycorrhizas was substantiated and no symplasmic connections were found between symbionts. Fluorescence studies demonstrated that 5,6-carboxyfluorescein was transported through the phloem into the roots of E. curvula, but remained predominantly in the root phloem. Unloading only occurred in optimal nutrient exchange areas of meristimatic lateral or apical growth regions. It was not possible, using fluorescence techniques and related equipment available, to conclusively establish if there were symplasmic connections between the mycobiont and its host or if bidirectional transfer of nutrients occurred at the same interface.
- Full Text:
- Date Issued: 2007
The role of arbuscular mycorrhizal fungi in the biotransformation of coal and application in dump rehabilitation
- Mukasa-Mugerwa, Thomas Tendo
- Authors: Mukasa-Mugerwa, Thomas Tendo
- Date: 2007
- Subjects: Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Fungi -- Biotechnology , Bermuda grass , Coal mines and mining -- Environmental aspects , Acid mine drainage
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3999 , http://hdl.handle.net/10962/d1004059 , Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Fungi -- Biotechnology , Bermuda grass , Coal mines and mining -- Environmental aspects , Acid mine drainage
- Description: Fundamental processes underpinning the biotransformation of coal by fungal biocatalysts have been intensively investigated, however, limited large-scale industrial applications using such systems have been reported. The un-anticipated sporadic growth of Cynodon dactylon on the surface of un-rehabilitated discard coal dumps has been noted and this was found to be coupled with the breakdown of coal into a humic soil-like material in the top 1.5 metres of the dumps. Extensive fungal growth was observed to be associated with the Cynodon dactylon root system and examination of plant roots indicated the presence of mycorrhizal fungi. Analysis of the Cynodon dactylon plant roots around which coal biotransformation was occurring confirmed the presence of arbuscular mycorrhizal colonisation with the species Glomus clarum, Paraglomus occultum, Gigaspora gigantea and Glomus mosseae identified to be associated with the plants. Further molecular characterisation of non-mycorrhizal rhizospheric fungi showed the presence of fungal species with coal-degrading capabilities that most likely played a role in the coal biotransformation observed. The discard coal dump environment was simulated in pot and column studies and coal biotransformation was reproduced, with this process enhanced by the addition of mycorrhizal and non-mycorrhizal rhizospheric fungal inocula to the environment. Mycorrhizal and non-mycorrhizal species in the inoculum were re-isolated from the simulated environment fulfilling a number of Koch’s postulates and indicating a causal role in the biotransformation of coal. An inversion of conventional mycorrhizal colonisation was demonstrated in this system with reduction in extraradicular presence and an increase in intracellular colonisation compared to soil controls. A descriptive model was formulated suggesting a two-part fungal system involving organic carbon and nutrient exchange between the plant, mycorrhizal fungi and non-mycorrhizal coal-degrading rhizospheric fungi ultimately resulting in the biotransformation of coal. The biotransformation observed was comparable to reports of “rock-eating fungi”. Results suggest that the biological degradation of coal in situ with the production of a soil-like substrate could provide a feasible method of discard coal dump rehabilitation as well as provide a humic-rich substrate that can be utilised in further industrial applications.
- Full Text:
- Date Issued: 2007
- Authors: Mukasa-Mugerwa, Thomas Tendo
- Date: 2007
- Subjects: Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Fungi -- Biotechnology , Bermuda grass , Coal mines and mining -- Environmental aspects , Acid mine drainage
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3999 , http://hdl.handle.net/10962/d1004059 , Vesicular-arbuscular mycorrhizas , Mycorrhizal fungi , Fungi -- Biotechnology , Bermuda grass , Coal mines and mining -- Environmental aspects , Acid mine drainage
- Description: Fundamental processes underpinning the biotransformation of coal by fungal biocatalysts have been intensively investigated, however, limited large-scale industrial applications using such systems have been reported. The un-anticipated sporadic growth of Cynodon dactylon on the surface of un-rehabilitated discard coal dumps has been noted and this was found to be coupled with the breakdown of coal into a humic soil-like material in the top 1.5 metres of the dumps. Extensive fungal growth was observed to be associated with the Cynodon dactylon root system and examination of plant roots indicated the presence of mycorrhizal fungi. Analysis of the Cynodon dactylon plant roots around which coal biotransformation was occurring confirmed the presence of arbuscular mycorrhizal colonisation with the species Glomus clarum, Paraglomus occultum, Gigaspora gigantea and Glomus mosseae identified to be associated with the plants. Further molecular characterisation of non-mycorrhizal rhizospheric fungi showed the presence of fungal species with coal-degrading capabilities that most likely played a role in the coal biotransformation observed. The discard coal dump environment was simulated in pot and column studies and coal biotransformation was reproduced, with this process enhanced by the addition of mycorrhizal and non-mycorrhizal rhizospheric fungal inocula to the environment. Mycorrhizal and non-mycorrhizal species in the inoculum were re-isolated from the simulated environment fulfilling a number of Koch’s postulates and indicating a causal role in the biotransformation of coal. An inversion of conventional mycorrhizal colonisation was demonstrated in this system with reduction in extraradicular presence and an increase in intracellular colonisation compared to soil controls. A descriptive model was formulated suggesting a two-part fungal system involving organic carbon and nutrient exchange between the plant, mycorrhizal fungi and non-mycorrhizal coal-degrading rhizospheric fungi ultimately resulting in the biotransformation of coal. The biotransformation observed was comparable to reports of “rock-eating fungi”. Results suggest that the biological degradation of coal in situ with the production of a soil-like substrate could provide a feasible method of discard coal dump rehabilitation as well as provide a humic-rich substrate that can be utilised in further industrial applications.
- Full Text:
- Date Issued: 2007
The presence and role of arbuscular mycorrhizal fungi in coastal sand dune systems
- Authors: Haller, Anjanette H. A
- Date: 2000
- Subjects: Mycorrhizas , Sand dune ecology -- South Africa , Mycorrhizal fungi
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4196 , http://hdl.handle.net/10962/d1003765 , Mycorrhizas , Sand dune ecology -- South Africa , Mycorrhizal fungi
- Description: Arbuscular mycorrhizas (AM) are mutually beneficial symbiotic associations between the roots of plants and certain Zygomycetous fungi. The role of AM fungi in coastal sand dunes has been explored in many parts of the world, though little work has been conducted in South African dune systems. This study aimed to investigate the presence and extent of mycorrhizal colonisation of a coastal sand dune in South Africa. The roots of five plant species (Scaevola plumieri, Arctotheca populifolia, Ipomoea pes-caprae, Ehrharta villosa and Chrysanthemoides monilifera) were sampled along a foredune profile at Old Woman's River in the Eastern Cape. These roots were assessed for the percentage mycorrhizal colonisation they supported. Spores extracted from the rhizosphere sand of each plant species were counted and identified to genus level. Results were related to seasonality and the position of the plants along the profile. All plant species were found to be mycorrhizal. Percentage colonisation ranged from 0-92%, depending on plant species and season. Mycorrhizal colonisation was generally highest in the winter months, and especially so in I pes-caprae and E. villosa. The extent of various mycorrhizal structures in root tissue varied between plant species. Spore numbers ranged from 0-48 spores 100g-1 sand with highest numbers occurring in winter. S. plumieri and A. populifolia were associated with greatest spore abundance. Four fungal genera (Glomus, Acaulospora, Scutellospora and Gigaspora) were identified. Distribution of these genera showed seasonal variations between plant species. A bioassay, using Sorghum, was conducted to test the inoculum potentials of sand from the Scaevola hummock and the IpomoealEhrharta dune. Highest percentage colonisation occurred in plants grown in the Scaevola sand, which also had the lowest root and shoot measurements. The bioassay confirmed that AM propagules are present and viable, even in the mobile sand of the foredune. This study showed that mycorrhizal colonisation and spore numbers varied seasonally, but that the extent of this was dependent on plant species. The position of plants along the foredune profile tended to be less important than plant species. It is thought that the growth cycle and rooting system of each plant species determines seasonal cycles and abundance of AM fungi. Variation within fungal populations probably also impacts on this. Knowledge of the presence and distribution of AM fungi in this system paves the way for more detailed studies which need to examine the role of these endophytes in South African sand dunes.
- Full Text:
- Date Issued: 2000
- Authors: Haller, Anjanette H. A
- Date: 2000
- Subjects: Mycorrhizas , Sand dune ecology -- South Africa , Mycorrhizal fungi
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4196 , http://hdl.handle.net/10962/d1003765 , Mycorrhizas , Sand dune ecology -- South Africa , Mycorrhizal fungi
- Description: Arbuscular mycorrhizas (AM) are mutually beneficial symbiotic associations between the roots of plants and certain Zygomycetous fungi. The role of AM fungi in coastal sand dunes has been explored in many parts of the world, though little work has been conducted in South African dune systems. This study aimed to investigate the presence and extent of mycorrhizal colonisation of a coastal sand dune in South Africa. The roots of five plant species (Scaevola plumieri, Arctotheca populifolia, Ipomoea pes-caprae, Ehrharta villosa and Chrysanthemoides monilifera) were sampled along a foredune profile at Old Woman's River in the Eastern Cape. These roots were assessed for the percentage mycorrhizal colonisation they supported. Spores extracted from the rhizosphere sand of each plant species were counted and identified to genus level. Results were related to seasonality and the position of the plants along the profile. All plant species were found to be mycorrhizal. Percentage colonisation ranged from 0-92%, depending on plant species and season. Mycorrhizal colonisation was generally highest in the winter months, and especially so in I pes-caprae and E. villosa. The extent of various mycorrhizal structures in root tissue varied between plant species. Spore numbers ranged from 0-48 spores 100g-1 sand with highest numbers occurring in winter. S. plumieri and A. populifolia were associated with greatest spore abundance. Four fungal genera (Glomus, Acaulospora, Scutellospora and Gigaspora) were identified. Distribution of these genera showed seasonal variations between plant species. A bioassay, using Sorghum, was conducted to test the inoculum potentials of sand from the Scaevola hummock and the IpomoealEhrharta dune. Highest percentage colonisation occurred in plants grown in the Scaevola sand, which also had the lowest root and shoot measurements. The bioassay confirmed that AM propagules are present and viable, even in the mobile sand of the foredune. This study showed that mycorrhizal colonisation and spore numbers varied seasonally, but that the extent of this was dependent on plant species. The position of plants along the foredune profile tended to be less important than plant species. It is thought that the growth cycle and rooting system of each plant species determines seasonal cycles and abundance of AM fungi. Variation within fungal populations probably also impacts on this. Knowledge of the presence and distribution of AM fungi in this system paves the way for more detailed studies which need to examine the role of these endophytes in South African sand dunes.
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- Date Issued: 2000
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