Sequencing, assembly and annotation of the mitochondrial and plastid genomes of Gelidium pristoides (Turner) Kützing from Kenton-on-Sea, South Africa
- Authors: Mangali, Sandisiwe
- Date: 2019
- Subjects: Gelidium -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/19109 , vital:39883
- Description: The genome is the complete set of an organism's hereditary information that contains all the information necessary for the functioning of that organism. Complete nuclear, mitochondrial and plastid DNA constitute the three main types of genomes which play interconnected roles in an organism. Genome sequencing enables researchers to understand the regulation and expression of the various genes and the proteins they encode. It allows researchers to extract and analyse genes of interests for a variety of studies including molecular, biotechnological, bioinformatics and conservation and evolutionary studies. Genome sequencing of Rhodophyta has received little attention. To date, no published studies are focusing on both whole genome sequencing and sequencing of the organellar genomes of Rhodophyta species found in along the South African coastline. This study focused on genome sequencing, assembly and annotation mitochondrial and plastid genomes of Gelidium pristoides. Gelidium pristoides was collected from Kenton-on-Sea and was morphologically identified at Rhodes University. Its genomic DNA was extracted using the Nucleospin® Plant II kit and quantified using Qubit 2.0, Nanodrop and 1% agarose gel electrophoresis. The Ion Plus Fragment Library kit was used for the preparation of a 600 bp library, which was sequenced in two separate runs through the Ion S5 platform. The produced reads were quality-controlled through the Ion Torrent server version 5.6. and assessed using the FASTQC program. The SPAdes version 3.11.1 assembler was used to assemble the quality-controlled reads, and the resultant genome assembly was quality-assessed using the QUAST 4.1 software. The mitochondrial genome was selected from the produced Gelidium pristoides draft genome using mitochondrial genomes of other Gelidiales as search queries on the local BLAST algorithm of the BioEdit software. Contigs matching the organellar genomes were ordered according to the mitochondrial genomes of other Gelidiales using the trial version of Geneious R11.12 software. The plastid genome was also selected following the same approach but using plastid genomes of Gelidium elegans and Gelidium vagum as search queries. Gaps observed in the organellar genomes were closed by amplification of the relevant gap using polymerase chain reaction with newly designed primers and Sanger sequencing. Open reading frames for both organellar genomes were annotated using the NCBI ORF-Finder and alignments obtained from BlastN and BlastX searches from the NCBI database, while the tRNAs and rRNAs were identified using the tRNAscan-SE1.21 vi and the RNAmmer 1.2 servers. The circular physical map of the mitochondrial genome was constructed using the CGView server. Lastly, in silico analysis of cytochrome c oxidase 3 and Heat Shock Protein 70 was performed using the PRIMO and the SWISS-MODEL pipelines respectively. Their phylogenies were analysed through Clustal omega and the trees viewed on TreeView 1.6.6 software. Qubit and Nanodrop genomic DNA qualification revealed A260/A280 and A230/A260 ratios of 1.81 and 1.52 respectively. The 1% agarose gel electrophoresis further confirmed the good quality of the genomic DNA used for library preparation and sequencing. Pre-assembly quality control of reads resulted in a total of 30 792 074 high-quality reads which were assembled into a total of 94140 contigs, making up an estimated genome length of 217.06 Mb. The largest contig covered up to 13.17 kb of the draft genome, and an N50 statistic value of 3.17 kb was obtained. The G.pristoides mitochondrial genome mapped into a circular molecule of 25012 bp, with an overall GC content of 31.04% and a total of 45 genes distributed into 20 tRNA-coding, 2 rRNAcoding genes and 23 protein-coding genes, mostly adopting the modified genetic code of Rhodophyta. The SecY and rps12 genes overlapped by 41 bp. This study presents a partial plastid genome composed of 89 (38%) fully annotated genes, of which 71 are protein-coding, and 18 are distributed among 15 tRNA-coding, 2 rRNA-coding and 1 RNaseP RNA-coding genes. Sixty-one (26%) partial protein-coding genes were predicted, while approximately 84 (36%) genes are not yet predicted. In silico analysis of the cytochrome c oxidase and heat shock protein 70 showed that the gene sequences obtained in this study and the resultant transcribed protein have sequences and structures that are similar to those from several other different species, thus validating the integrity of the genome sequences. This study provides genomic data necessary for understanding the genomic constituent of G.pristoides and serve as a foundation for studies of individual genes and for resolving evolutionary relationships.
- Full Text:
- Date Issued: 2019
- Authors: Mangali, Sandisiwe
- Date: 2019
- Subjects: Gelidium -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/19109 , vital:39883
- Description: The genome is the complete set of an organism's hereditary information that contains all the information necessary for the functioning of that organism. Complete nuclear, mitochondrial and plastid DNA constitute the three main types of genomes which play interconnected roles in an organism. Genome sequencing enables researchers to understand the regulation and expression of the various genes and the proteins they encode. It allows researchers to extract and analyse genes of interests for a variety of studies including molecular, biotechnological, bioinformatics and conservation and evolutionary studies. Genome sequencing of Rhodophyta has received little attention. To date, no published studies are focusing on both whole genome sequencing and sequencing of the organellar genomes of Rhodophyta species found in along the South African coastline. This study focused on genome sequencing, assembly and annotation mitochondrial and plastid genomes of Gelidium pristoides. Gelidium pristoides was collected from Kenton-on-Sea and was morphologically identified at Rhodes University. Its genomic DNA was extracted using the Nucleospin® Plant II kit and quantified using Qubit 2.0, Nanodrop and 1% agarose gel electrophoresis. The Ion Plus Fragment Library kit was used for the preparation of a 600 bp library, which was sequenced in two separate runs through the Ion S5 platform. The produced reads were quality-controlled through the Ion Torrent server version 5.6. and assessed using the FASTQC program. The SPAdes version 3.11.1 assembler was used to assemble the quality-controlled reads, and the resultant genome assembly was quality-assessed using the QUAST 4.1 software. The mitochondrial genome was selected from the produced Gelidium pristoides draft genome using mitochondrial genomes of other Gelidiales as search queries on the local BLAST algorithm of the BioEdit software. Contigs matching the organellar genomes were ordered according to the mitochondrial genomes of other Gelidiales using the trial version of Geneious R11.12 software. The plastid genome was also selected following the same approach but using plastid genomes of Gelidium elegans and Gelidium vagum as search queries. Gaps observed in the organellar genomes were closed by amplification of the relevant gap using polymerase chain reaction with newly designed primers and Sanger sequencing. Open reading frames for both organellar genomes were annotated using the NCBI ORF-Finder and alignments obtained from BlastN and BlastX searches from the NCBI database, while the tRNAs and rRNAs were identified using the tRNAscan-SE1.21 vi and the RNAmmer 1.2 servers. The circular physical map of the mitochondrial genome was constructed using the CGView server. Lastly, in silico analysis of cytochrome c oxidase 3 and Heat Shock Protein 70 was performed using the PRIMO and the SWISS-MODEL pipelines respectively. Their phylogenies were analysed through Clustal omega and the trees viewed on TreeView 1.6.6 software. Qubit and Nanodrop genomic DNA qualification revealed A260/A280 and A230/A260 ratios of 1.81 and 1.52 respectively. The 1% agarose gel electrophoresis further confirmed the good quality of the genomic DNA used for library preparation and sequencing. Pre-assembly quality control of reads resulted in a total of 30 792 074 high-quality reads which were assembled into a total of 94140 contigs, making up an estimated genome length of 217.06 Mb. The largest contig covered up to 13.17 kb of the draft genome, and an N50 statistic value of 3.17 kb was obtained. The G.pristoides mitochondrial genome mapped into a circular molecule of 25012 bp, with an overall GC content of 31.04% and a total of 45 genes distributed into 20 tRNA-coding, 2 rRNAcoding genes and 23 protein-coding genes, mostly adopting the modified genetic code of Rhodophyta. The SecY and rps12 genes overlapped by 41 bp. This study presents a partial plastid genome composed of 89 (38%) fully annotated genes, of which 71 are protein-coding, and 18 are distributed among 15 tRNA-coding, 2 rRNA-coding and 1 RNaseP RNA-coding genes. Sixty-one (26%) partial protein-coding genes were predicted, while approximately 84 (36%) genes are not yet predicted. In silico analysis of the cytochrome c oxidase and heat shock protein 70 showed that the gene sequences obtained in this study and the resultant transcribed protein have sequences and structures that are similar to those from several other different species, thus validating the integrity of the genome sequences. This study provides genomic data necessary for understanding the genomic constituent of G.pristoides and serve as a foundation for studies of individual genes and for resolving evolutionary relationships.
- Full Text:
- Date Issued: 2019
Investigating the use of Arbuscular Mycorrhizas and Plant Growth Promoting Bacteria to improve the drought tolerance of maize (Zea mays L.)
- Authors: Moore, Nicolle Maureen
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54587 , vital:26591
- Description: Maize (Zea mays L.) is a direct staple food crop in Africa and remains an essential component of global food security, with maize crops accounting for over 60% of the total harvested area of annual food crops. Stress caused by drought and high soil salinity limits crop growth and productivity more than any other single environmental factor, with grain yield reductions up to 76% depending on the severity of the drought and the plant growth stage. Arbuscular mycorrhizal (AM) fungi and Plant Growth Promotion Rhizobacteria (PGPR) have previously been shown to improve tolerance of plants to drought stress through a number of chemical and physiological processes. The aim of this investigation was to determine whether mycorrhizal fungi and rhizobacteria adapted to drought and saline conditions and possessing plant growth promoting (PGP) traits were able to stimulate plant growth responses when applied to Zea mays seeds growing under greenhouse conditions Bacterial isolates selected were tolerant to concentrations of NaCl up to 600 mM and maintained 50% growth at low water potentials (-1.44 MPa). They were positive for Indole Acetic Acid (IAA) production, phosphate solubilisation and secretion of siderophores. Bacterial isolates showing plant growth promoting potential were identified using 16S rDNA gene sequencing as Achromobacter xylosoxidans strains A8 and C54 and Klebsiella oxytoca strain M1. Mixed inoculum was prepared from indigenous communities of mycorrhizas in soils sampled from the Cerebos Salt Pan and the Kalahari Desert. Mycorrhizal diversity was investigated using 454-Pyrosequencing which revealed that the community composition was dominated by species in the Ambispora, Glomus and Paraglomus genera with a rare component represented by species in the Redeckera, Archaeospora and Geosiphon genera. Microscopic examination of plant roots at the end of the trial revealed the presence of diagnostic mycorrhizal structures within the root cells, confirming that colonization was successful. Plant growth response to microbial inoculation was assessed by monitoring changes in plant photosynthetic capacity over the duration of a 7 week pot trial. A significant difference in photosynthetic and biomass data was observed between drought and well-watered groups but no mycorrhizal or bacterial treatment effect was evident within the groups, despite the high levels of colonization by mycorrhizas. These results suggest that the beneficial effects of mycorrhizal colonization may be primarily attributed to improved nutrient and mineral uptake in conditions where nutrients are limiting, resulting in improved growth. The improved growth may then have secondary effects on the plant‟s ability to withstand drought. Having controlled for nutrient deficiency, it was not evident in this study that mycorrhizal fungi were able to stimulate a change in plant physiology and confer drought tolerance under the conditions imposed.
- Full Text:
- Date Issued: 2016
- Authors: Moore, Nicolle Maureen
- Date: 2016
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54587 , vital:26591
- Description: Maize (Zea mays L.) is a direct staple food crop in Africa and remains an essential component of global food security, with maize crops accounting for over 60% of the total harvested area of annual food crops. Stress caused by drought and high soil salinity limits crop growth and productivity more than any other single environmental factor, with grain yield reductions up to 76% depending on the severity of the drought and the plant growth stage. Arbuscular mycorrhizal (AM) fungi and Plant Growth Promotion Rhizobacteria (PGPR) have previously been shown to improve tolerance of plants to drought stress through a number of chemical and physiological processes. The aim of this investigation was to determine whether mycorrhizal fungi and rhizobacteria adapted to drought and saline conditions and possessing plant growth promoting (PGP) traits were able to stimulate plant growth responses when applied to Zea mays seeds growing under greenhouse conditions Bacterial isolates selected were tolerant to concentrations of NaCl up to 600 mM and maintained 50% growth at low water potentials (-1.44 MPa). They were positive for Indole Acetic Acid (IAA) production, phosphate solubilisation and secretion of siderophores. Bacterial isolates showing plant growth promoting potential were identified using 16S rDNA gene sequencing as Achromobacter xylosoxidans strains A8 and C54 and Klebsiella oxytoca strain M1. Mixed inoculum was prepared from indigenous communities of mycorrhizas in soils sampled from the Cerebos Salt Pan and the Kalahari Desert. Mycorrhizal diversity was investigated using 454-Pyrosequencing which revealed that the community composition was dominated by species in the Ambispora, Glomus and Paraglomus genera with a rare component represented by species in the Redeckera, Archaeospora and Geosiphon genera. Microscopic examination of plant roots at the end of the trial revealed the presence of diagnostic mycorrhizal structures within the root cells, confirming that colonization was successful. Plant growth response to microbial inoculation was assessed by monitoring changes in plant photosynthetic capacity over the duration of a 7 week pot trial. A significant difference in photosynthetic and biomass data was observed between drought and well-watered groups but no mycorrhizal or bacterial treatment effect was evident within the groups, despite the high levels of colonization by mycorrhizas. These results suggest that the beneficial effects of mycorrhizal colonization may be primarily attributed to improved nutrient and mineral uptake in conditions where nutrients are limiting, resulting in improved growth. The improved growth may then have secondary effects on the plant‟s ability to withstand drought. Having controlled for nutrient deficiency, it was not evident in this study that mycorrhizal fungi were able to stimulate a change in plant physiology and confer drought tolerance under the conditions imposed.
- Full Text:
- Date Issued: 2016
Assessing the genetic diversity of catface rockcod epinephelus andersoni in the subtropical Western Indian Ocean and modelling the effects of climate change on their distribution
- Authors: Coppinger, Christine Rose
- Date: 2014
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54499 , vital:26570
- Description: The catface rockcod Epinephelus andersoni is a range-restricted species that is endemic to the southeast coast of Africa from Quissico in Mozambique (subtropical) to Knysna in South Africa (warmtemperate). Its complex life-history, long-lived nature and high residency make E. andersoni potentially vulnerable to over-exploitation. Epinephelus andersoni is an important fishery species and has shown signs of depletion. Due to inadequate information necessary for management and conservation, further research is vital, particularly in the face of potentially significant climatic changes which could put further pressure on E. andersoni. The aim of the study was to provide information for the management of E. andersoni, with considerations for the possibly detrimental effects of future climate change. The objectives of this study were to describe the genetic structure and diversity of E. andersoni and to determine possible range shifts of E. andersoni with future changes in sea surface temperature. Genetic samples were collected throughout the distribution of E. andersoni. Standard DNA extraction and PCR using universal primers were conducted and nuclear (RPS7-1) and mitochondrial (cytochrome b) data were analysed to determine genetic diversity. A combination of nuclear and mitochondrial markers was used to ensure that the results were robust. RPS7-1 haplotype diversity was high (0.801) and an AMOVA on the RPS7-1 data showed significantly high among group variation (ΦCT = 0.204, p < 0.05) between five groups: 1. Quissico to Inhaca; 2. Cape Vidal to Port Edward; 3 Port St Johns to Coffee Bay; 4. Mbashe; 5. Port Alfred. This geographic structuring could be attributed to low gene flow across barriers such as the Port Alfred upwelling cell, the Mozambique Channel eddies and smaller more localised upwelling cells such as the Port St Johns cell. The cytochrome b results contrastingly indicate low haplotype diversity (0.309) and no differentiation (ΦCT = 0.265, p = 0.074) between groups and support the hypothesis of a historical population bottleneck. This may be due to an unusually slower mutation rate of the cytochrome b region than the RPS7-1 region, resulting in the RPS7-1 data showing a more recent picture of diversification. To complement the genetic results, niche modelling techniques were used to determine range shifts of E. andersoni with future temperature trends using species distribution and climatic data. The model illustrated a contraction of the E. andersoni distribution as well as future intensification of various upwelling cells along the south-east African coast including the Port Alfred upwelling cell. Due to the low gene flow across these barriers this intensification could decrease the resilience of E. andersoni, as its range becomes more limited with global change. The genetic data and modelling results combined provide useful information on which to base future fisheries management.
- Full Text:
- Date Issued: 2014
- Authors: Coppinger, Christine Rose
- Date: 2014
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/54499 , vital:26570
- Description: The catface rockcod Epinephelus andersoni is a range-restricted species that is endemic to the southeast coast of Africa from Quissico in Mozambique (subtropical) to Knysna in South Africa (warmtemperate). Its complex life-history, long-lived nature and high residency make E. andersoni potentially vulnerable to over-exploitation. Epinephelus andersoni is an important fishery species and has shown signs of depletion. Due to inadequate information necessary for management and conservation, further research is vital, particularly in the face of potentially significant climatic changes which could put further pressure on E. andersoni. The aim of the study was to provide information for the management of E. andersoni, with considerations for the possibly detrimental effects of future climate change. The objectives of this study were to describe the genetic structure and diversity of E. andersoni and to determine possible range shifts of E. andersoni with future changes in sea surface temperature. Genetic samples were collected throughout the distribution of E. andersoni. Standard DNA extraction and PCR using universal primers were conducted and nuclear (RPS7-1) and mitochondrial (cytochrome b) data were analysed to determine genetic diversity. A combination of nuclear and mitochondrial markers was used to ensure that the results were robust. RPS7-1 haplotype diversity was high (0.801) and an AMOVA on the RPS7-1 data showed significantly high among group variation (ΦCT = 0.204, p < 0.05) between five groups: 1. Quissico to Inhaca; 2. Cape Vidal to Port Edward; 3 Port St Johns to Coffee Bay; 4. Mbashe; 5. Port Alfred. This geographic structuring could be attributed to low gene flow across barriers such as the Port Alfred upwelling cell, the Mozambique Channel eddies and smaller more localised upwelling cells such as the Port St Johns cell. The cytochrome b results contrastingly indicate low haplotype diversity (0.309) and no differentiation (ΦCT = 0.265, p = 0.074) between groups and support the hypothesis of a historical population bottleneck. This may be due to an unusually slower mutation rate of the cytochrome b region than the RPS7-1 region, resulting in the RPS7-1 data showing a more recent picture of diversification. To complement the genetic results, niche modelling techniques were used to determine range shifts of E. andersoni with future temperature trends using species distribution and climatic data. The model illustrated a contraction of the E. andersoni distribution as well as future intensification of various upwelling cells along the south-east African coast including the Port Alfred upwelling cell. Due to the low gene flow across these barriers this intensification could decrease the resilience of E. andersoni, as its range becomes more limited with global change. The genetic data and modelling results combined provide useful information on which to base future fisheries management.
- Full Text:
- Date Issued: 2014
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
The molecular microbial ecology of sulfate reduction in the Rhodes BioSURE process
- Authors: Chauke, Chesa Gift
- Date: 2002
- Subjects: Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4082 , http://hdl.handle.net/10962/d1007475 , Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Description: The research reported here investigated the use of a Baffle Reactor in order to study aspects of the biological sulfur cycle, where a floating sulfur biofilm formation occurs and where complex organic compounds provide electron donor sources. The development of a laboratory-scale Baffle Reactor model system satisfied the requirements for sulfate reducing bacterial biomass growth and sulfur biofilm formation. Since relatively little is known about the microbial ecology of floating sulfur biofilm systems, this study was undertaken to describe the sulfate reducing sludge population of the system together with its performance. A combination of culture- and molecular-based techniques were applied in this study in order to investigate the microbial ecology of the sulfate-reducing bacteria component of the system. These techniques enabled the identification and the analysis of the distribution of different sulfate reducing bacterial strains found within the sludge bioreactors. Strains isolated from the sludge were characterised based on culture appearance, gram staining and scanning electron microscopy morphology. Molecular methods based on the PCR-amplified 16S rRNA including denaturing gradient gel electrophoresis were employed in order to characterise sulfate-reducing bacteria within the reactors. Three novel Gram negative sulfate-reducing bacteria strains were isolated from the sludge population. Strains isolated were tentatively named Desulfomonas rhodensis, Desulfomonas makanaiensis, and Clostridium sulforhodensis. Results obtained from the Baffle Reactor showed that three dominant species were isolated from the DNA extracted from the whole bacterial population by peR. Three of these were similar to those mentioned above. The presence of these three novel unidentified species suggest that there are a range of other novel organisms involved in sulfate reduction processes.
- Full Text:
- Date Issued: 2002
- Authors: Chauke, Chesa Gift
- Date: 2002
- Subjects: Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4082 , http://hdl.handle.net/10962/d1007475 , Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Description: The research reported here investigated the use of a Baffle Reactor in order to study aspects of the biological sulfur cycle, where a floating sulfur biofilm formation occurs and where complex organic compounds provide electron donor sources. The development of a laboratory-scale Baffle Reactor model system satisfied the requirements for sulfate reducing bacterial biomass growth and sulfur biofilm formation. Since relatively little is known about the microbial ecology of floating sulfur biofilm systems, this study was undertaken to describe the sulfate reducing sludge population of the system together with its performance. A combination of culture- and molecular-based techniques were applied in this study in order to investigate the microbial ecology of the sulfate-reducing bacteria component of the system. These techniques enabled the identification and the analysis of the distribution of different sulfate reducing bacterial strains found within the sludge bioreactors. Strains isolated from the sludge were characterised based on culture appearance, gram staining and scanning electron microscopy morphology. Molecular methods based on the PCR-amplified 16S rRNA including denaturing gradient gel electrophoresis were employed in order to characterise sulfate-reducing bacteria within the reactors. Three novel Gram negative sulfate-reducing bacteria strains were isolated from the sludge population. Strains isolated were tentatively named Desulfomonas rhodensis, Desulfomonas makanaiensis, and Clostridium sulforhodensis. Results obtained from the Baffle Reactor showed that three dominant species were isolated from the DNA extracted from the whole bacterial population by peR. Three of these were similar to those mentioned above. The presence of these three novel unidentified species suggest that there are a range of other novel organisms involved in sulfate reduction processes.
- Full Text:
- Date Issued: 2002
Ichthyofaunal community structures in different types of Eastern Cape estuaries
- Authors: Vorwerk, Paul D
- Date: 2001
- Subjects: Estuarine fishes -- South Africa -- Eastern Cape , Estuarine fishes -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5377 , http://hdl.handle.net/10962/d1016241
- Description: This investigation had three major objectives; firstly to document the estuarine fishes from several poorly studied Eastern Cape estuaries, secondly, to identify similarities and differences in the fish communities from permanently open and intermittently open estuaries, and thirdly, to establish which physico-chemical or environmental variables are responsible for the structuring of these communities. The study area incorporated 10 estuaries along a 70 km long stretch of the Eastern Cape coastline between the towns of Seafield (33° 32' 42" S, 27° 03' 05" E) in the south-east and Hamburg (33° 16' 45" S, 27° 29' 50" E) in the north-west. The systems investigated included two permanently open estuaries and eight intermittently open systems of varying sizes. Overall fish abundance and species richness in the study area revealed no significant seasonal variation. A contrast between open and closed estuaries was evident in the dominance of species from different estuarine dependence categories. Open estuaries comprised mainly category Ib, lla and IIb species, with a high proportion of category IV species. Closed estuaries were dominated by category IIa species, with few category IV species represented. Clear longitudinal trends in relative abundance were demonstrated for individual species. Margalefs species richness index for seine net catches, and a combination of the seine and gill net data, were significantly correlated with catchment size, mean annual runoff, estuarine area and linear length. Fish assemblages in the permanently open and intermittently open estuaries were significantly different (p<0.001) when analysed on a community and density basis. Similarly, the communities in large and small intermittently open estuaries differed significantly on a community (p=0.01) and density basis (p<0.001). These differences where caused by changes in abundance of the dominant estuarine resident and marine migrant species. The environmental variable that was dominant in accounting for these differences was the estuary mouth status. Estuarine resident species had smaller overall body lengths when compared with the marine migrant species. In addition, the estuarine resident species had smaller modal size classes in the closed estuaries when compared with the open estuaries. These differences may be linked to the foraging strategies of the species, as well as food availability and different levels of predation in the different estuary types. The marine migrant species did not show any trends in this respect although the closed systems had the largest individuals for all four dominant marine species. This finding may be linked to the inability of these species to breed in closed estuaries, thus channelling reproductive energy resources into somatic growth. In addition, prolonged periods of mouth closure prevent the emigration of large individuals back to the sea. Otter trawling was conducted in fiye estuaries and captured mostly demersal species. The otter trawl data confirmed the seine net community analysis, with significantly different communities identified in the permanently open and intermittently open estuaries (p=0.02), as well as in the small and large intermittently open estuaries (p=0.03). The environmental variables responsible for these community differences were mouth status and variables associated with estuarine size. The identification of estuary mouth status by this study as the most influential environmental factor is compounded by this "ariable affecting or being indicative of numerous other physico-chemical features. The large differences in the fish communities in different estuary types indicate the importance of each estuary type to various species. This should be taken into account before allowing any form of commercial fishing in these important nursery areas.
- Full Text:
- Date Issued: 2001
- Authors: Vorwerk, Paul D
- Date: 2001
- Subjects: Estuarine fishes -- South Africa -- Eastern Cape , Estuarine fishes -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5377 , http://hdl.handle.net/10962/d1016241
- Description: This investigation had three major objectives; firstly to document the estuarine fishes from several poorly studied Eastern Cape estuaries, secondly, to identify similarities and differences in the fish communities from permanently open and intermittently open estuaries, and thirdly, to establish which physico-chemical or environmental variables are responsible for the structuring of these communities. The study area incorporated 10 estuaries along a 70 km long stretch of the Eastern Cape coastline between the towns of Seafield (33° 32' 42" S, 27° 03' 05" E) in the south-east and Hamburg (33° 16' 45" S, 27° 29' 50" E) in the north-west. The systems investigated included two permanently open estuaries and eight intermittently open systems of varying sizes. Overall fish abundance and species richness in the study area revealed no significant seasonal variation. A contrast between open and closed estuaries was evident in the dominance of species from different estuarine dependence categories. Open estuaries comprised mainly category Ib, lla and IIb species, with a high proportion of category IV species. Closed estuaries were dominated by category IIa species, with few category IV species represented. Clear longitudinal trends in relative abundance were demonstrated for individual species. Margalefs species richness index for seine net catches, and a combination of the seine and gill net data, were significantly correlated with catchment size, mean annual runoff, estuarine area and linear length. Fish assemblages in the permanently open and intermittently open estuaries were significantly different (p<0.001) when analysed on a community and density basis. Similarly, the communities in large and small intermittently open estuaries differed significantly on a community (p=0.01) and density basis (p<0.001). These differences where caused by changes in abundance of the dominant estuarine resident and marine migrant species. The environmental variable that was dominant in accounting for these differences was the estuary mouth status. Estuarine resident species had smaller overall body lengths when compared with the marine migrant species. In addition, the estuarine resident species had smaller modal size classes in the closed estuaries when compared with the open estuaries. These differences may be linked to the foraging strategies of the species, as well as food availability and different levels of predation in the different estuary types. The marine migrant species did not show any trends in this respect although the closed systems had the largest individuals for all four dominant marine species. This finding may be linked to the inability of these species to breed in closed estuaries, thus channelling reproductive energy resources into somatic growth. In addition, prolonged periods of mouth closure prevent the emigration of large individuals back to the sea. Otter trawling was conducted in fiye estuaries and captured mostly demersal species. The otter trawl data confirmed the seine net community analysis, with significantly different communities identified in the permanently open and intermittently open estuaries (p=0.02), as well as in the small and large intermittently open estuaries (p=0.03). The environmental variables responsible for these community differences were mouth status and variables associated with estuarine size. The identification of estuary mouth status by this study as the most influential environmental factor is compounded by this "ariable affecting or being indicative of numerous other physico-chemical features. The large differences in the fish communities in different estuary types indicate the importance of each estuary type to various species. This should be taken into account before allowing any form of commercial fishing in these important nursery areas.
- Full Text:
- Date Issued: 2001
Aspects of the anaerobic digestion of wattle tannins
- Authors: Hurlow, Elton Lloyd
- Date: 1987
- Subjects: Tanneries -- Waste disposal
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4300 , http://hdl.handle.net/10962/d1004909
- Description: The aqueous extract from the bark of the black wattle tree, (Acacia mearnsii De Wild)(1), is used extensively in the tanning industry for the production of sole and other heavy leathers as well as in lighter leathers and skins. The commercial extract is available in an easily dissolvable spray dried form and in deference to its genus name Mimosaceae is referred to in the trade as Mimosa(3). Mimosa extract consists primarily of high molecular weight polymeric flavonoid units which also constitutes the active tanning ingredient. Lower phenolics, gums and sugars or the so called "non-tannins" are present as impurities and contribute up to 40% to (3 ) the total mass of the extract.
- Full Text:
- Date Issued: 1987
- Authors: Hurlow, Elton Lloyd
- Date: 1987
- Subjects: Tanneries -- Waste disposal
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4300 , http://hdl.handle.net/10962/d1004909
- Description: The aqueous extract from the bark of the black wattle tree, (Acacia mearnsii De Wild)(1), is used extensively in the tanning industry for the production of sole and other heavy leathers as well as in lighter leathers and skins. The commercial extract is available in an easily dissolvable spray dried form and in deference to its genus name Mimosaceae is referred to in the trade as Mimosa(3). Mimosa extract consists primarily of high molecular weight polymeric flavonoid units which also constitutes the active tanning ingredient. Lower phenolics, gums and sugars or the so called "non-tannins" are present as impurities and contribute up to 40% to (3 ) the total mass of the extract.
- Full Text:
- Date Issued: 1987
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