Ericoid mycorrhizal fungi and potential for inoculation of commercial berry species (Vaccinium corymbosium L.)
- Authors: Bizabani, Christine
- Date: 2011
- Subjects: Ericaceae , Mycorrhizas , Fynbos
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4136 , http://hdl.handle.net/10962/d1016127
- Description: Ericaceous plants are the richest growth form of the fynbos vegetation of South Africa. The fynbos is characterized by highly leached acidic soils, low mineral nutrients and climatically it is a winter rainfall and dry summer region. Ericoid mycorrhizal fungi associate with Erica species enhancing their ability to access essential nutrients for survival under unfavourable growth conditions. The aim of this study was to select local Ericaceae plant species and to isolate, identify and characterize the ericoid endophytes and assess these isolates as potential inocula for commercial berry species. Two ericaceous plants Erica cerinthoides L. and Erica demmissa Klotzsch ex Benth. were identified from the Mountain Drive area of Grahamstown, Eastern Cape. Root staining was used to confirm the mycorrhizal status of both plants. Hyphal coils typical of ericoid association were observed within the epidermal cells of the hair roots under a light microscope. The endophytes were successfully isolated in pure culture on 2% malt extract agar (MEA) and modified Fontana medium. Cultural morphology and microscopy were used for initial identification. Two slow growing isolates were selected. These isolates were further subjected to molecular identification; extracted DNA was amplified using ITS1 and ITS4 fungal primers. The rDNA gene internal transcriber spacer (ITS) was then sequenced and analyzed by comparison to sequences in the GenBank. On the basis of percentage sequence identity Lachnum Retz. species and Cadophora Lagerb. & Melin species were identified as the ericoid endophytes of E. cerinthoides and E. demmissa respectively. The optimum growth parameters of the fungal isolates were determined in 2% MEA incubated at varying temperatures and pH. It was established that both species had optimum growth at 27⁰C and pH 5. The Ericaceae species are sometimes found in metal contaminated sites were ericoid fungi have been proved to alleviate toxicity of their host. The fungal isolates were grown in increasing concentration of Cu²⁺ and Zn²⁺ in 2% MEA. The growth of Lachnum species decreased with increasing Zn²⁺ ions above 2.7 mM while Cadophora species showed a change in morphology and also decreased in growth with increased ion concentration. However there were no significant differences recorded in the growth of Cadophora and Lachnum species on increasing Cu²⁺ concentration. Lachnum and Cadophora isolates were formulated into a semi solid inoculum and inoculated onto micropropagated Vaccinni corymbosum L. plantlets of 5 different varieties. Colonization was low for all varieties, Elliott and Brightwell varieties recorded the highest colonization of 35% and 31% respectively. Lachnum species infected roots showed potential ericoid structures while the Cadophora inoculated plantlets had hyphal coils within the cortical cells typical of ericoid mycorrhizas. Inoculation significantly enhanced the shoot growth of Brightwell and Elliott varieties. The Chandler variety inoculated with Lachnum species showed improved shoot dry weight. The Bluecrop and Elliott varieties inoculated with Cadophora and Lachnum accumulated more root biomass. All inoculated Bluecrop plantlets had an improved canopy growth index. Brightwell plantlets inoculated with Lachnum species also had an enhanced canopy growth index. The growth responses were variable within varieties and between varieties. Treatments with the Cadophora and Lachnum have shown potential in the promotion of growth of the Blueberry species. The findings indicate the need to conduct trials under conditions which simulate the commercial growth conditions so as explore the optimum potential of the isolates.
- Full Text:
- Date Issued: 2011
- Authors: Bizabani, Christine
- Date: 2011
- Subjects: Ericaceae , Mycorrhizas , Fynbos
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4136 , http://hdl.handle.net/10962/d1016127
- Description: Ericaceous plants are the richest growth form of the fynbos vegetation of South Africa. The fynbos is characterized by highly leached acidic soils, low mineral nutrients and climatically it is a winter rainfall and dry summer region. Ericoid mycorrhizal fungi associate with Erica species enhancing their ability to access essential nutrients for survival under unfavourable growth conditions. The aim of this study was to select local Ericaceae plant species and to isolate, identify and characterize the ericoid endophytes and assess these isolates as potential inocula for commercial berry species. Two ericaceous plants Erica cerinthoides L. and Erica demmissa Klotzsch ex Benth. were identified from the Mountain Drive area of Grahamstown, Eastern Cape. Root staining was used to confirm the mycorrhizal status of both plants. Hyphal coils typical of ericoid association were observed within the epidermal cells of the hair roots under a light microscope. The endophytes were successfully isolated in pure culture on 2% malt extract agar (MEA) and modified Fontana medium. Cultural morphology and microscopy were used for initial identification. Two slow growing isolates were selected. These isolates were further subjected to molecular identification; extracted DNA was amplified using ITS1 and ITS4 fungal primers. The rDNA gene internal transcriber spacer (ITS) was then sequenced and analyzed by comparison to sequences in the GenBank. On the basis of percentage sequence identity Lachnum Retz. species and Cadophora Lagerb. & Melin species were identified as the ericoid endophytes of E. cerinthoides and E. demmissa respectively. The optimum growth parameters of the fungal isolates were determined in 2% MEA incubated at varying temperatures and pH. It was established that both species had optimum growth at 27⁰C and pH 5. The Ericaceae species are sometimes found in metal contaminated sites were ericoid fungi have been proved to alleviate toxicity of their host. The fungal isolates were grown in increasing concentration of Cu²⁺ and Zn²⁺ in 2% MEA. The growth of Lachnum species decreased with increasing Zn²⁺ ions above 2.7 mM while Cadophora species showed a change in morphology and also decreased in growth with increased ion concentration. However there were no significant differences recorded in the growth of Cadophora and Lachnum species on increasing Cu²⁺ concentration. Lachnum and Cadophora isolates were formulated into a semi solid inoculum and inoculated onto micropropagated Vaccinni corymbosum L. plantlets of 5 different varieties. Colonization was low for all varieties, Elliott and Brightwell varieties recorded the highest colonization of 35% and 31% respectively. Lachnum species infected roots showed potential ericoid structures while the Cadophora inoculated plantlets had hyphal coils within the cortical cells typical of ericoid mycorrhizas. Inoculation significantly enhanced the shoot growth of Brightwell and Elliott varieties. The Chandler variety inoculated with Lachnum species showed improved shoot dry weight. The Bluecrop and Elliott varieties inoculated with Cadophora and Lachnum accumulated more root biomass. All inoculated Bluecrop plantlets had an improved canopy growth index. Brightwell plantlets inoculated with Lachnum species also had an enhanced canopy growth index. The growth responses were variable within varieties and between varieties. Treatments with the Cadophora and Lachnum have shown potential in the promotion of growth of the Blueberry species. The findings indicate the need to conduct trials under conditions which simulate the commercial growth conditions so as explore the optimum potential of the isolates.
- Full Text:
- Date Issued: 2011
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 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.
- Full Text:
- Date Issued: 2000
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