Applied phylogeography : mapping the genetic resource of Honeybush across the Cape Floristic Region
- Authors: Galuszynski, Nicholas C
- Date: 2020
- Subjects: Phylogeography -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/46771 , vital:39655
- Description: Aim: Honeybush tea is a herbal infusion made from members of the Cape Floristic Region (CFR) endemic Cyclopia Vent. Consting of 21 extant species, six are anthropogenically redistributed across the CFR for Honeybush cultivation. However, with no information regarding the distribution and levels of genetic diversity among wild Honeybush populations, anthropogenic translocation of cultivated genotypes may risk disrupting natural genetic diversity patterns. In this thesis, an applied phylogeographic approach is used to provide baseline insights into the spatial structuring of Honeybush genetic diversity. Location: The Cape Floristic Region (CFR), located along the southern Cape of South Africa. Methods: Spatial structuring of evolutionary lineages within Cyclopia are explored at a genus level using data generated from the combination of Anchored Hybrid Enrichment library preparation and high through put sequencing. A High Resolution Melt analysis (HRM) toolkit is then developed to screen haplotype variation across three non-coding chloroplast loci. This HRM toolkit coupled with haplotype confirmation Sanger sequencing is then applied to describe the spatial structuring of genetic diversity in wild populations for two Honeybush species (C. intermedia and C. subternata) and to compare genetic diversity among wild and cultivated populations of three Honeybush species (C. intermedia, C. subternata, and C. longifolia). A reanalysis of published phylogeographic data focusing on CFR plant taxa is used to test if phylogeographic breaks occur across the boundaries among adjacent Centers of Endemism (CoEs). Finally, the low-copy nuclear data set generated via AHE and high throughput sequencing is summarized. Results: Phylogeographic structuring was detected at the inter- and intraspecific levels in Cyclopia. The genus level analysis revealed a strong phylogenetic split between western CFR and eastern CFR endemic species in the nuclear and chloroplast genomes. At the intraspecific level, HRM proved to be a high throughput and accurate tool for haplotype detection, revealing phylogeographic structuring of genetic diversity in C.intermediaand C.subternata. This genetic diversity and structuring was not detected in cultivated Honeybush populations, which appear to have originated from a small number of founding individuals and have experienced a genetic bottleneck. Spatial structuring of genetic lineages is common among CFR plant taxa and the reanalysis of existing data found a significant trend for phylogeographic breaks tooccuracrosstheboundariesamongadjacentCoEs. Finally, over200000baseswere sequenced across 445 low-copy nuclear loci for 14 Cyclopia species. Conclusions: This thesis provides important baseline information on the spatial distribution of wild Honeybush genetic diversity. Phylogeographic structuring is present in wild Honeybush populations. This however is not accounted for in cultivated populations, which may place wild genetic diversity at risk of being disrupted if geneflow among wild and cultivated populations occurs. The results from this thesis should therefore be used to develop precautionary guidelines for the anthropogenic redistribution of Honeybush genetic material for cultivation. Furthermore, the novel insights into regional patterns of phylogeohraphic structuring of plant populations should encourage more efficient and hypothesis driven sampling designs in future phylogeographic work in the CFR. The high throughput sequence data generated in this thesis should transform Cyclopia to a model organism for the study of molecular evolution in the CFR.
- Full Text:
- Date Issued: 2020
- Authors: Galuszynski, Nicholas C
- Date: 2020
- Subjects: Phylogeography -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/46771 , vital:39655
- Description: Aim: Honeybush tea is a herbal infusion made from members of the Cape Floristic Region (CFR) endemic Cyclopia Vent. Consting of 21 extant species, six are anthropogenically redistributed across the CFR for Honeybush cultivation. However, with no information regarding the distribution and levels of genetic diversity among wild Honeybush populations, anthropogenic translocation of cultivated genotypes may risk disrupting natural genetic diversity patterns. In this thesis, an applied phylogeographic approach is used to provide baseline insights into the spatial structuring of Honeybush genetic diversity. Location: The Cape Floristic Region (CFR), located along the southern Cape of South Africa. Methods: Spatial structuring of evolutionary lineages within Cyclopia are explored at a genus level using data generated from the combination of Anchored Hybrid Enrichment library preparation and high through put sequencing. A High Resolution Melt analysis (HRM) toolkit is then developed to screen haplotype variation across three non-coding chloroplast loci. This HRM toolkit coupled with haplotype confirmation Sanger sequencing is then applied to describe the spatial structuring of genetic diversity in wild populations for two Honeybush species (C. intermedia and C. subternata) and to compare genetic diversity among wild and cultivated populations of three Honeybush species (C. intermedia, C. subternata, and C. longifolia). A reanalysis of published phylogeographic data focusing on CFR plant taxa is used to test if phylogeographic breaks occur across the boundaries among adjacent Centers of Endemism (CoEs). Finally, the low-copy nuclear data set generated via AHE and high throughput sequencing is summarized. Results: Phylogeographic structuring was detected at the inter- and intraspecific levels in Cyclopia. The genus level analysis revealed a strong phylogenetic split between western CFR and eastern CFR endemic species in the nuclear and chloroplast genomes. At the intraspecific level, HRM proved to be a high throughput and accurate tool for haplotype detection, revealing phylogeographic structuring of genetic diversity in C.intermediaand C.subternata. This genetic diversity and structuring was not detected in cultivated Honeybush populations, which appear to have originated from a small number of founding individuals and have experienced a genetic bottleneck. Spatial structuring of genetic lineages is common among CFR plant taxa and the reanalysis of existing data found a significant trend for phylogeographic breaks tooccuracrosstheboundariesamongadjacentCoEs. Finally, over200000baseswere sequenced across 445 low-copy nuclear loci for 14 Cyclopia species. Conclusions: This thesis provides important baseline information on the spatial distribution of wild Honeybush genetic diversity. Phylogeographic structuring is present in wild Honeybush populations. This however is not accounted for in cultivated populations, which may place wild genetic diversity at risk of being disrupted if geneflow among wild and cultivated populations occurs. The results from this thesis should therefore be used to develop precautionary guidelines for the anthropogenic redistribution of Honeybush genetic material for cultivation. Furthermore, the novel insights into regional patterns of phylogeohraphic structuring of plant populations should encourage more efficient and hypothesis driven sampling designs in future phylogeographic work in the CFR. The high throughput sequence data generated in this thesis should transform Cyclopia to a model organism for the study of molecular evolution in the CFR.
- Full Text:
- Date Issued: 2020
Phylogeny and phylogeography of dominant South African surf diatoms
- Authors: Du Preez, Debbie Patricia
- Date: 2017
- Subjects: Phylogeography -- South Africa , Diatoms -- South Africa Phylogeny -- Molecular aspects DNA -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/29794 , vital:30778
- Description: Genetic studies of diatoms are lagging behind those on other organisms. The reasons for this are, among others, that many species are difficult to culture (some surf diatoms in particular), single cells contain only a small amount of DNA, and selection of single cells from environmental samples is difficult. Globally, there are only a few species of surf diatom: Anaulus australis Drebes et Schultz, Asterionellopsis glacialis (Castracane) Round, Asterionellopsis socialis (Lewin & Norris) Crawford & Gardner, Attheya armata (West) Crawford, Aulacodiscus petersii Ehrenberg, Aulacodiscus johnsonii Arnott, Aulacodiscus kittonii Arnott ex Ralfs and Aulacodiscus africanus Cottam. These species occur exclusively in the surf zone with the exception of A. glacialis, which is also found in coastal and oceanic waters. The overall aim of this project was to investigate the phylogeny or the phylogeography of the two dominant South African surf diatoms. In order to do this, it was necessary to develop methods for extracting DNA from environmental surf diatom patch samples. This method was then used in the investigation into the phylogeography of Anaulus australis along the South African coastal zone. Due to other recent work into diatom phylogeny and phylogeography there was also the opportunity to compare the genetics of the South African surf diatom Asterionellopsis found at Sundays River Beach to localities recently sampled and sequenced around the globe in order to identify the species found in South Africa. The direct method was successful for DNA extraction from Anaulus australis cells preserved using either isopropanol, ethanol or glutaraldehyde (provided the samples were not older than two years), but not formalin. Extraction of DNA from Asterionellopsis glacialis s.l. cells was successful with the direct method for ethanol-preserved samples, but the extraction from Attheya armata cells required some modifications be added to the method. Sequencing of the ITS region of Anaulus australis cells from along the southern coast of South Africa indicated connectivity between the various populations rather than the expected isolation of populations in the surf zones of the log-spiral bays. Comparison of the ITS and RbcL regions of the species of Asterionellopsis found at the Sundays River Beach indicated that it is Asterionellopsis lenisilicea which is a species only ii recorded from Canada to date and which has only recently been discovered and recognised as a new species within the genus. This genetic identification was supported by morphological measurements of frustules.
- Full Text:
- Date Issued: 2017
- Authors: Du Preez, Debbie Patricia
- Date: 2017
- Subjects: Phylogeography -- South Africa , Diatoms -- South Africa Phylogeny -- Molecular aspects DNA -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/29794 , vital:30778
- Description: Genetic studies of diatoms are lagging behind those on other organisms. The reasons for this are, among others, that many species are difficult to culture (some surf diatoms in particular), single cells contain only a small amount of DNA, and selection of single cells from environmental samples is difficult. Globally, there are only a few species of surf diatom: Anaulus australis Drebes et Schultz, Asterionellopsis glacialis (Castracane) Round, Asterionellopsis socialis (Lewin & Norris) Crawford & Gardner, Attheya armata (West) Crawford, Aulacodiscus petersii Ehrenberg, Aulacodiscus johnsonii Arnott, Aulacodiscus kittonii Arnott ex Ralfs and Aulacodiscus africanus Cottam. These species occur exclusively in the surf zone with the exception of A. glacialis, which is also found in coastal and oceanic waters. The overall aim of this project was to investigate the phylogeny or the phylogeography of the two dominant South African surf diatoms. In order to do this, it was necessary to develop methods for extracting DNA from environmental surf diatom patch samples. This method was then used in the investigation into the phylogeography of Anaulus australis along the South African coastal zone. Due to other recent work into diatom phylogeny and phylogeography there was also the opportunity to compare the genetics of the South African surf diatom Asterionellopsis found at Sundays River Beach to localities recently sampled and sequenced around the globe in order to identify the species found in South Africa. The direct method was successful for DNA extraction from Anaulus australis cells preserved using either isopropanol, ethanol or glutaraldehyde (provided the samples were not older than two years), but not formalin. Extraction of DNA from Asterionellopsis glacialis s.l. cells was successful with the direct method for ethanol-preserved samples, but the extraction from Attheya armata cells required some modifications be added to the method. Sequencing of the ITS region of Anaulus australis cells from along the southern coast of South Africa indicated connectivity between the various populations rather than the expected isolation of populations in the surf zones of the log-spiral bays. Comparison of the ITS and RbcL regions of the species of Asterionellopsis found at the Sundays River Beach indicated that it is Asterionellopsis lenisilicea which is a species only ii recorded from Canada to date and which has only recently been discovered and recognised as a new species within the genus. This genetic identification was supported by morphological measurements of frustules.
- Full Text:
- Date Issued: 2017
Comparative phylogeography of five swallowtail butterfly species (Lepidoptera: Papilionidae) in South Africa : ecological and taxonomic implications.
- Authors: Neef, Götz-Georg
- Date: 2014
- Subjects: Swallowtail butterflies , Papilionidae -- South Africa , Phylogeography -- South Africa , Swallowtail butterflies -- Effect of habitat modification on , Biodiversity -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4266 , http://hdl.handle.net/10962/d1013574
- Description: With current biota under constant threat of extinction, it is important to ascertain where and how biological diversity is generated and partitioned. Phylogeographic studies can assist in the identification of places and processes that indicate the origin and maintenance of biodiversity. Forest fragmentation has a big effect on local extinction and loss of genetic diversity of forest-restricted taxa, along with divergence and speciation of forest biota. This study aims to understand the effects of these processes on a number of forest-dwelling butterflies using a comparative phylogeographic approach. Mitochondrial DNA of five different Papilio species with different degrees of forest specificity was analysed using phylogenetic methods. In addition, the subspecific taxonomy of P. ophidicephalus was investigated using morphometrics of discal spots on the wings and nuclear DNA analysis along with mitochondrial DNA analysis. The results show that the forest-restricted species (P. ophidicephalus and P. echerioides) have more genetic structure and less genetic diversity than the more generalist species (P. dardanus, P. demodocus and P. nireus). This could be due to inbreeding depression and bottlenecks caused by forest fragmentation. As forest patches become smaller, the population size is affected and that causes a loss in genetic diversity, and increasing habitat fragmentation disrupts gene flow. The intraspecific taxonomy of P. ophidicephalus is far from revealed. However, this study shows there is evidence for the different subspecies when comparing morphological results and genetic results. From the evidence provided here it is suggested that P. ophidicephalus should be divided into two separate species rather than five subspecies.
- Full Text:
- Date Issued: 2014
- Authors: Neef, Götz-Georg
- Date: 2014
- Subjects: Swallowtail butterflies , Papilionidae -- South Africa , Phylogeography -- South Africa , Swallowtail butterflies -- Effect of habitat modification on , Biodiversity -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4266 , http://hdl.handle.net/10962/d1013574
- Description: With current biota under constant threat of extinction, it is important to ascertain where and how biological diversity is generated and partitioned. Phylogeographic studies can assist in the identification of places and processes that indicate the origin and maintenance of biodiversity. Forest fragmentation has a big effect on local extinction and loss of genetic diversity of forest-restricted taxa, along with divergence and speciation of forest biota. This study aims to understand the effects of these processes on a number of forest-dwelling butterflies using a comparative phylogeographic approach. Mitochondrial DNA of five different Papilio species with different degrees of forest specificity was analysed using phylogenetic methods. In addition, the subspecific taxonomy of P. ophidicephalus was investigated using morphometrics of discal spots on the wings and nuclear DNA analysis along with mitochondrial DNA analysis. The results show that the forest-restricted species (P. ophidicephalus and P. echerioides) have more genetic structure and less genetic diversity than the more generalist species (P. dardanus, P. demodocus and P. nireus). This could be due to inbreeding depression and bottlenecks caused by forest fragmentation. As forest patches become smaller, the population size is affected and that causes a loss in genetic diversity, and increasing habitat fragmentation disrupts gene flow. The intraspecific taxonomy of P. ophidicephalus is far from revealed. However, this study shows there is evidence for the different subspecies when comparing morphological results and genetic results. From the evidence provided here it is suggested that P. ophidicephalus should be divided into two separate species rather than five subspecies.
- Full Text:
- Date Issued: 2014
Phylogeography and epifauna of two intertidal seaweeds on the coast of South Africa
- Authors: Mmonwa, Lucas Kolobe
- Date: 2009
- Subjects: Phylogeography -- South Africa , Marine algae -- South Africa , Red algae -- South Africa , Gelidium -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5738 , http://hdl.handle.net/10962/d1005424 , Phylogeography -- South Africa , Marine algae -- South Africa , Red algae -- South Africa , Gelidium -- South Africa
- Description: Southern African biogeographic boundaries delimit the phylogeographic distribution of some coastal and estuarine invertebrates. This study investigated the impact of these boundaries on the phylogeographic distribution of two intertidal red seaweeds, Gelidium pristoides and Hypnea spicifera using the mitochondrial Cox2-3 spacer and the nuclear ITS1 regions. G. pristoides spores have short distance-dispersal, while long distance-dispersal is more likely in H. spicifera via spores and drifting fertile thallus fragments. Both markers revealed a south-western and south-eastern lineage within G. pristoides but the breaks between lineages do not coincide with any recognised biogeographic limits. The Cox2-3 spacer revealed a boundary between the two lineages at the Alexandria Coastal Dunefield (ACD) and ITS1 at the Gamtoos-Van Stadens Dunefields (GVD) which is approximately 80km west of the ACD. The minor difference between the two markers regarding location of the phylogeographic boundary is probably due to the dating differences between the two dunefields. The ACD as developed currently is superimposed on the ancient dunefields which formed during the Pleistocene, coinciding with the Cox2-3 spacer sequences divergence which dates back 500,000 - 580,000 years. The GVD formed during the Holocene (6,500 - 4,000 years ago), coinciding with the ITS1 sequences divergence which dates 4,224 - 4,928 years ago. Thus, these phylogeographic boundaries probably appeared without the influence of biogeographic boundaries, but rather due to the lack of suitable habitat in the dunefields, coupled with short dispersal-distances of the spores. Analysis of the ITS1 and Cox2-3 spacer regions in H. spicifera revealed that the species is characterized by uniform genetic structure along the coastline. This reflects the species`s potential for long range expansion as it inhabits both the intertidal and subtidal zones; and this presumably leads to high gene flow among populations. The ITS1 sequences showed minimal genetic variation of one substitution between the gametophyte and tetrasporophyte generations within H. spicifera. This suggests the predominance of asexual reproduction, which reduces gene flow and fixes alleles between generations. ANOSIM and Bray-Curtis cluster analyses showed scale-dependant variation in the abundances of epifauna (mainly amphipod, isopod, mollusc and polychaete species) on G. pristoides. At small local (within site) and large (among sites) scales, there were weak and no structure in epifaunal abundances respectively. However, at larger, biogeographic scales, samples from the same biogeographic region tended to be clustered together. Thus, there was a group containing predominantly south coast samples and a group containing east coast samples mixed with the remaining south coast samples. Such scale-dependant variation in epifaunal abundances is probably due to the effects of factors driving species richness at small local (within site) scales (e.g. wave exposure, seaweed biomass) and at larger, biogeographic scales (e.g. surface sea temperature). Moreover, at very small (individual samples) scales; there was no correlation between epifauna composition and genotype of the seaweed. Seaweed samples characterized by distinct ITS1 or Cox2-3 spacer sequences did not show any significant differences in epifaunal composition. Although the distributional pattern of the epifaunal community observed at large biogeographic scale is not clear, it seems to be associated with the biogeographic regions. However, phylogeographic distribution of Gelidium pristoides is not connected to biogeographic regions. Thus, at larger, biogeographic scales, there is no correlation between phylogeographic distribution of G. pristoides and distribution of the associated fauna
- Full Text:
- Date Issued: 2009
- Authors: Mmonwa, Lucas Kolobe
- Date: 2009
- Subjects: Phylogeography -- South Africa , Marine algae -- South Africa , Red algae -- South Africa , Gelidium -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5738 , http://hdl.handle.net/10962/d1005424 , Phylogeography -- South Africa , Marine algae -- South Africa , Red algae -- South Africa , Gelidium -- South Africa
- Description: Southern African biogeographic boundaries delimit the phylogeographic distribution of some coastal and estuarine invertebrates. This study investigated the impact of these boundaries on the phylogeographic distribution of two intertidal red seaweeds, Gelidium pristoides and Hypnea spicifera using the mitochondrial Cox2-3 spacer and the nuclear ITS1 regions. G. pristoides spores have short distance-dispersal, while long distance-dispersal is more likely in H. spicifera via spores and drifting fertile thallus fragments. Both markers revealed a south-western and south-eastern lineage within G. pristoides but the breaks between lineages do not coincide with any recognised biogeographic limits. The Cox2-3 spacer revealed a boundary between the two lineages at the Alexandria Coastal Dunefield (ACD) and ITS1 at the Gamtoos-Van Stadens Dunefields (GVD) which is approximately 80km west of the ACD. The minor difference between the two markers regarding location of the phylogeographic boundary is probably due to the dating differences between the two dunefields. The ACD as developed currently is superimposed on the ancient dunefields which formed during the Pleistocene, coinciding with the Cox2-3 spacer sequences divergence which dates back 500,000 - 580,000 years. The GVD formed during the Holocene (6,500 - 4,000 years ago), coinciding with the ITS1 sequences divergence which dates 4,224 - 4,928 years ago. Thus, these phylogeographic boundaries probably appeared without the influence of biogeographic boundaries, but rather due to the lack of suitable habitat in the dunefields, coupled with short dispersal-distances of the spores. Analysis of the ITS1 and Cox2-3 spacer regions in H. spicifera revealed that the species is characterized by uniform genetic structure along the coastline. This reflects the species`s potential for long range expansion as it inhabits both the intertidal and subtidal zones; and this presumably leads to high gene flow among populations. The ITS1 sequences showed minimal genetic variation of one substitution between the gametophyte and tetrasporophyte generations within H. spicifera. This suggests the predominance of asexual reproduction, which reduces gene flow and fixes alleles between generations. ANOSIM and Bray-Curtis cluster analyses showed scale-dependant variation in the abundances of epifauna (mainly amphipod, isopod, mollusc and polychaete species) on G. pristoides. At small local (within site) and large (among sites) scales, there were weak and no structure in epifaunal abundances respectively. However, at larger, biogeographic scales, samples from the same biogeographic region tended to be clustered together. Thus, there was a group containing predominantly south coast samples and a group containing east coast samples mixed with the remaining south coast samples. Such scale-dependant variation in epifaunal abundances is probably due to the effects of factors driving species richness at small local (within site) scales (e.g. wave exposure, seaweed biomass) and at larger, biogeographic scales (e.g. surface sea temperature). Moreover, at very small (individual samples) scales; there was no correlation between epifauna composition and genotype of the seaweed. Seaweed samples characterized by distinct ITS1 or Cox2-3 spacer sequences did not show any significant differences in epifaunal composition. Although the distributional pattern of the epifaunal community observed at large biogeographic scale is not clear, it seems to be associated with the biogeographic regions. However, phylogeographic distribution of Gelidium pristoides is not connected to biogeographic regions. Thus, at larger, biogeographic scales, there is no correlation between phylogeographic distribution of G. pristoides and distribution of the associated fauna
- Full Text:
- Date Issued: 2009
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