Comparative study of the feeding damage caused by the South African biotypes of the Russian wheat aphid (Diuraphis noxia Kurdjumov) on resistant and non-resistant lines of barley (Hordeum vulgare L.)
- Authors: Jimoh, Mahboob Adekilekun
- Date: 2011
- Subjects: Aphids Russian wheat aphid -- Research -- South Africa Barley -- Diseases and pests -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4201 , http://hdl.handle.net/10962/d1003770
- Description: Cereal crop productivity is hampered when these plants are infested by phloem feeding aphids. A great deal of research has been carried out with the direct aim of a clearer understanding of the mechanism involved in the interaction between aphids and their host plants. Research has directly or indirectly been geared towards enhanced plant productivity and achieving sustainable agriculture. Barley (Hordeum vulgare L.) is an important small grain crop in South Africa, whose crop performance is negatively affected by fluctuations in weather patterns as well as by agricultural pests. One of the insect pests infesting barley is the Russian wheat aphid, Diuraphis noxia Kurdjumov (RWA), of which the two South African biotypes, codenamed RWASA1 and RWASA2, were studied in this thesis. During dry spells, RWA infestation becomes a more serious threat to barley productivity. Resistant plants have been used to combat RWA infestation of small grains. In South Africa, 27 RWA-resistant wheat cultivars are currently used in commercial cultivation, but no resistant barley lines have, unfortunately, been developed, in spite of this grain’s significant economic importance. This informed the study in this thesis, and this interest particularly focussed on three RWA-resistant lines developed by the USDA, testing their performance against South African RWA biotypes, for possible adaptation to South Africa. The aim was thus to examine the plant-aphid interactions, aphid breeding rates, plant damage and sustainability, evidence of resistance or tolerance and finally potential performance under elevated CO2 (a very real climate change threat). Two major avenues of research were undertaken. The first aspect involved examination of structural and functional damage caused by RWASA1 and RWASA2 on the three resistant and a non-resistant line. Aphid population growth and damage symptoms (chlorosis and leaf roll) of infestation by these aphid biotypes were evaluated. This was followed by a structural and functional approach in which the effects of feeding on the transport systems (phloem and xylem) of barley were investigated. Fluorescence microscopy techniques (using aniline blue fluorochrome, a specific stain for callose and 5,6-CFDA, a phloem-mobile probe) were applied to investigate the feeding-related damage caused by the aphids, through an examination of wound callose formation and related to this, the resultant reduction in phloem transport capacity. Transmission electron microscopy (TEM) techniques provided evidence of the extent of the feeding-related cell damage. The second aspect involved a study of the effect of changing CO2 concentrations ([CO2]) on the resistant and susceptible barley cultivars to feeding by the two RWA biotypes. Leaves of plants grown at ambient and two elevated levels of [CO2] were analysed to investigate the effect of changing [CO2] on biomass, leaf nitrogen content and C:N ratio of control (uninfested) and infested plants. The population growth studies showed that the populations of the two RWA biotypes as well as bird cherry-oat aphid (BCA, Rhopalosiphum padi L.) increased substantially on the four barley lines. BCA was included here, as it had been the subject of several previous studies. RWASA2 bred faster than RWASA1 on all lines. The breeding rates of the two RWA biotypes were both suppressed and at near-equivalent levels on the three resistant lines, compared to the non-resistant PUMA. This suggests that the resistant lines possessed an antibiosis resistance mechanism against the feeding aphids. Feeding by the aphids manifested in morphological damage symptoms of chlorosis and leaf roll. The two biotypes inflicted severe chlorosis and leaf roll on the non-resistant PUMA. In the resistant plants, leaf rolling was more severe because of RWASA2 feeding compared to RWASA1 feeding. In contrast, chlorosis symptoms were more severe during RWASA1 feeding than was the case with RWASA2 feeding. Investigation of the effect of aphid feeding on the plants showed that callose was deposited within 24h and that this increased with longer feeding exposure. Wound callose distribution is more extensive in the non-resistant PUMA than in the resistant plants. RWASA2 feeding on the resistant lines caused deposition of more callose than was evident with RWASA1 feeding. During long-term feeding, it was evident that variation in the intensity and amount of wound callose was visible in the longitudinal and transverse veins of the resistant plants. Of the three STARS plants, STARS-9301B had the least callose. Interestingly, wound callose occurred in both resistant and non-resistant plants, in sharp contrast to what has been reported on resistant wheat cultivars that were developed in South Africa. The relative reduction in the wound callose deposited in the resistant line, when compared to the non-resistant lines, suggests the presence of a mechanism in the resistant lines, which may prevent excessive callose formation. Alternatively, the mechanism may stimulate callose breakdown. RWASA2 feeding on the resistant lines deposited more wound callose than RWASA1 feeding. This evidence supports the hypothesis that RWASA2 is a resistance breaking and more aggressive feeder than RWASA1 is; and further underscores the urgent need for development of RWA-resistant barley cultivars. The ultrastructural investigation of the feeding damage showed that the two biotypes caused extensive vascular damage in non-resistant plants. There was extensive and severe cell disruption and often obliteration of cell structure of the vascular parenchyma, xylem and phloem elements. In sharp contrast, among the resistant plants, feeding-related cell damage appeared to be substantially reduced compared to the non-resistant PUMA. Low frequency of damaged cells indicated that majority of the cell components of the vascular tissues were intact and presumed functional. There was evidence of salivary material lining the secondary walls of the vascular tissue, which resulted in severe damage. Within xylem vessels, saliva material impregnated half-bordered pit pairs between the vessels and adjacent xylem parenchyma. This is believed to prevent solute exchange through this interface, thereby inducing leaf stress and vi leaf roll. A notable finding is that RWASA2 effectively induced more cell damage to vascular tissues in the resistant lines than did RWASA1. In general the experimental evidence (see Chapter 5) suggests that the resistant lines are possibly more tolerant (or able to cope with) to RWA feeding. Evidence for this is the reduction of wound callose and at the TEM level, a comparatively less obvious cell damage in the resistant lines, which suggests that they possess antibiosis and tolerance capacity. The apparent reduction of feeding-related cell damage from the TEM study confirmed the disruptive action of the feeding aphids in experiments using the phloem-mobile probe, 5,6-CF. Results showed that feeding by RWASA1 and RWASA2 reducedthe transport functionality of the phloem in all cases, but that RWASA2 feeding caused a more obvious reduction in the rate and distance that 5,6-carboxyfluorescein was transported, than did RWASA1. Investigation of the effect of changing [CO2] on the barley cultivars showed that in the absence of aphids and under elevated CO2 conditions, the plants grew more vigorously. In this series of experiments, the infested plants suffered significant reduction in biomass under ambient (as was expected) and under the two elevated CO2 regimes. Biomass loss was greater at elevated CO2 than under ambient [CO2]. The infested nonresistant PUMA plants showed a more significant biomass loss than did the resistant cultivars. Clearly, the benefits derived from elevated CO2 enrichment was thus redirected to the now-advantaged aphids. Uninfested vii plants showed an increase in leaf nitrogen under the experimental conditions. However, feeding aphids depleted leaf nitrogen content and this was more apparent on plants exposed to RWASA2 than was the case with RWASA1. The end result of this was that C:N ratio of infested plants were higher than uninfested plants. Clearly, the faster breeding rates of the aphids at elevated CO2 caused depletion of N and a resultant deficiency exacerbated chlorosis as well as leaf rolling due to the higher aphid population density under elevated CO2 than at ambient. By 28 days after infestation (DAI), majority of the plants exposed to enriched CO2 treatments had died. A major finding here was thus that although this study demonstrated that elevated CO2 resulted in an increase in biomass, this was detrimentally offset in plants infested by the aphids, with a decline in biomass and loss of functionality leading to plant death at 28DAI. The overriding conclusion from this study is a clear signal that the twin effects of CO2 enrichment (a feature of current climate change) and aphid infestations may precipitate potential grain shortages. A disastrous food security threat looms.
- Full Text:
- Date Issued: 2011
- Authors: Jimoh, Mahboob Adekilekun
- Date: 2011
- Subjects: Aphids Russian wheat aphid -- Research -- South Africa Barley -- Diseases and pests -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4201 , http://hdl.handle.net/10962/d1003770
- Description: Cereal crop productivity is hampered when these plants are infested by phloem feeding aphids. A great deal of research has been carried out with the direct aim of a clearer understanding of the mechanism involved in the interaction between aphids and their host plants. Research has directly or indirectly been geared towards enhanced plant productivity and achieving sustainable agriculture. Barley (Hordeum vulgare L.) is an important small grain crop in South Africa, whose crop performance is negatively affected by fluctuations in weather patterns as well as by agricultural pests. One of the insect pests infesting barley is the Russian wheat aphid, Diuraphis noxia Kurdjumov (RWA), of which the two South African biotypes, codenamed RWASA1 and RWASA2, were studied in this thesis. During dry spells, RWA infestation becomes a more serious threat to barley productivity. Resistant plants have been used to combat RWA infestation of small grains. In South Africa, 27 RWA-resistant wheat cultivars are currently used in commercial cultivation, but no resistant barley lines have, unfortunately, been developed, in spite of this grain’s significant economic importance. This informed the study in this thesis, and this interest particularly focussed on three RWA-resistant lines developed by the USDA, testing their performance against South African RWA biotypes, for possible adaptation to South Africa. The aim was thus to examine the plant-aphid interactions, aphid breeding rates, plant damage and sustainability, evidence of resistance or tolerance and finally potential performance under elevated CO2 (a very real climate change threat). Two major avenues of research were undertaken. The first aspect involved examination of structural and functional damage caused by RWASA1 and RWASA2 on the three resistant and a non-resistant line. Aphid population growth and damage symptoms (chlorosis and leaf roll) of infestation by these aphid biotypes were evaluated. This was followed by a structural and functional approach in which the effects of feeding on the transport systems (phloem and xylem) of barley were investigated. Fluorescence microscopy techniques (using aniline blue fluorochrome, a specific stain for callose and 5,6-CFDA, a phloem-mobile probe) were applied to investigate the feeding-related damage caused by the aphids, through an examination of wound callose formation and related to this, the resultant reduction in phloem transport capacity. Transmission electron microscopy (TEM) techniques provided evidence of the extent of the feeding-related cell damage. The second aspect involved a study of the effect of changing CO2 concentrations ([CO2]) on the resistant and susceptible barley cultivars to feeding by the two RWA biotypes. Leaves of plants grown at ambient and two elevated levels of [CO2] were analysed to investigate the effect of changing [CO2] on biomass, leaf nitrogen content and C:N ratio of control (uninfested) and infested plants. The population growth studies showed that the populations of the two RWA biotypes as well as bird cherry-oat aphid (BCA, Rhopalosiphum padi L.) increased substantially on the four barley lines. BCA was included here, as it had been the subject of several previous studies. RWASA2 bred faster than RWASA1 on all lines. The breeding rates of the two RWA biotypes were both suppressed and at near-equivalent levels on the three resistant lines, compared to the non-resistant PUMA. This suggests that the resistant lines possessed an antibiosis resistance mechanism against the feeding aphids. Feeding by the aphids manifested in morphological damage symptoms of chlorosis and leaf roll. The two biotypes inflicted severe chlorosis and leaf roll on the non-resistant PUMA. In the resistant plants, leaf rolling was more severe because of RWASA2 feeding compared to RWASA1 feeding. In contrast, chlorosis symptoms were more severe during RWASA1 feeding than was the case with RWASA2 feeding. Investigation of the effect of aphid feeding on the plants showed that callose was deposited within 24h and that this increased with longer feeding exposure. Wound callose distribution is more extensive in the non-resistant PUMA than in the resistant plants. RWASA2 feeding on the resistant lines caused deposition of more callose than was evident with RWASA1 feeding. During long-term feeding, it was evident that variation in the intensity and amount of wound callose was visible in the longitudinal and transverse veins of the resistant plants. Of the three STARS plants, STARS-9301B had the least callose. Interestingly, wound callose occurred in both resistant and non-resistant plants, in sharp contrast to what has been reported on resistant wheat cultivars that were developed in South Africa. The relative reduction in the wound callose deposited in the resistant line, when compared to the non-resistant lines, suggests the presence of a mechanism in the resistant lines, which may prevent excessive callose formation. Alternatively, the mechanism may stimulate callose breakdown. RWASA2 feeding on the resistant lines deposited more wound callose than RWASA1 feeding. This evidence supports the hypothesis that RWASA2 is a resistance breaking and more aggressive feeder than RWASA1 is; and further underscores the urgent need for development of RWA-resistant barley cultivars. The ultrastructural investigation of the feeding damage showed that the two biotypes caused extensive vascular damage in non-resistant plants. There was extensive and severe cell disruption and often obliteration of cell structure of the vascular parenchyma, xylem and phloem elements. In sharp contrast, among the resistant plants, feeding-related cell damage appeared to be substantially reduced compared to the non-resistant PUMA. Low frequency of damaged cells indicated that majority of the cell components of the vascular tissues were intact and presumed functional. There was evidence of salivary material lining the secondary walls of the vascular tissue, which resulted in severe damage. Within xylem vessels, saliva material impregnated half-bordered pit pairs between the vessels and adjacent xylem parenchyma. This is believed to prevent solute exchange through this interface, thereby inducing leaf stress and vi leaf roll. A notable finding is that RWASA2 effectively induced more cell damage to vascular tissues in the resistant lines than did RWASA1. In general the experimental evidence (see Chapter 5) suggests that the resistant lines are possibly more tolerant (or able to cope with) to RWA feeding. Evidence for this is the reduction of wound callose and at the TEM level, a comparatively less obvious cell damage in the resistant lines, which suggests that they possess antibiosis and tolerance capacity. The apparent reduction of feeding-related cell damage from the TEM study confirmed the disruptive action of the feeding aphids in experiments using the phloem-mobile probe, 5,6-CF. Results showed that feeding by RWASA1 and RWASA2 reducedthe transport functionality of the phloem in all cases, but that RWASA2 feeding caused a more obvious reduction in the rate and distance that 5,6-carboxyfluorescein was transported, than did RWASA1. Investigation of the effect of changing [CO2] on the barley cultivars showed that in the absence of aphids and under elevated CO2 conditions, the plants grew more vigorously. In this series of experiments, the infested plants suffered significant reduction in biomass under ambient (as was expected) and under the two elevated CO2 regimes. Biomass loss was greater at elevated CO2 than under ambient [CO2]. The infested nonresistant PUMA plants showed a more significant biomass loss than did the resistant cultivars. Clearly, the benefits derived from elevated CO2 enrichment was thus redirected to the now-advantaged aphids. Uninfested vii plants showed an increase in leaf nitrogen under the experimental conditions. However, feeding aphids depleted leaf nitrogen content and this was more apparent on plants exposed to RWASA2 than was the case with RWASA1. The end result of this was that C:N ratio of infested plants were higher than uninfested plants. Clearly, the faster breeding rates of the aphids at elevated CO2 caused depletion of N and a resultant deficiency exacerbated chlorosis as well as leaf rolling due to the higher aphid population density under elevated CO2 than at ambient. By 28 days after infestation (DAI), majority of the plants exposed to enriched CO2 treatments had died. A major finding here was thus that although this study demonstrated that elevated CO2 resulted in an increase in biomass, this was detrimentally offset in plants infested by the aphids, with a decline in biomass and loss of functionality leading to plant death at 28DAI. The overriding conclusion from this study is a clear signal that the twin effects of CO2 enrichment (a feature of current climate change) and aphid infestations may precipitate potential grain shortages. A disastrous food security threat looms.
- Full Text:
- Date Issued: 2011
The genetic diversity and conservation biology of the rare terrestrial snail genus Prestonella
- Authors: Fearon, Janine Lee
- Date: 2011
- Subjects: Snails -- South Africa , Snails -- Conservation -- South Africa , Snails -- Variation -- South Africa , Biodiversity -- South Africa , Snails -- Genetics -- South Africa , Snails -- Habitat -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4191 , http://hdl.handle.net/10962/d1003760 , Snails -- South Africa , Snails -- Conservation -- South Africa , Snails -- Variation -- South Africa , Biodiversity -- South Africa , Snails -- Genetics -- South Africa , Snails -- Habitat -- South Africa
- Description: Prestonella bowkeri and Prestonella nuptialis are montane specialists endemic to the southern Great Escarpment of South Africa. Phylogeographic analyses of these species based on mitochondrial markers CO1 and 16S reveal extremely high levels of divergence between populations indicating a lack of gene flow between populations. This is not surprising, because P. nuptialis and P. bowkeri have limited dispersal capacity, low vagility, a highly fragmented distribution and are habitat specialists that are restricted to isolated mesic refugia associated with waterfalls and montane seepages. A relaxed Bayesian clock estimate suggests that populations diverged from one another during the mid-late Miocene (12.5-7 MYA) which coincides with the modern trends of seasonal aridity which began during the Miocene. This result should be viewed with caution because the rates used are at best imprecise estimates of mutation rates in snails. There is no clear dichotomy between the two species and P. bowkeri is paraphyletic with respect to P. nuptialis, as a consequence the taxonomy is unclear. Due to the high levels of sequence divergence between populations they may be considered as evolutionary significant units (ESU’s). An assessment of haplotype diversity (h) and nucleotide diversity (π) reveals that populations in the western part of the Great Escarpment are more genetically depauperate than populations in the east. Correlations between genetic diversity and climatic variables show that genetically depauperate populations are found in areas that have lower annual rainfall, less reliable rainfall and higher potential evaporation, all factors associated with a drier, less mesic environment that increases the chances of a population bottleneck. This indicates that a shift towards a more arid environment may be a driver of genetic erosion. Historical climate change may thus have affected the amount and distribution of genetic diversity across the Great Escarpment since the Miocene. This has serious future implications for the survival of Prestonella. With predicted increase in global temperatures, climate change in South Africa is likely to result in range contraction and an eastward range shift for many species in the drier central and western areas (Erasmus et al. 2002) and regions along the Great Escarpment are likely to become more arid. Prestonella populations found living on inselbergs along the Great Escarpment are already restricted to site specific watercourses and seepages. An increase in the periods between stream flow, and increasing rainfall variability and mean annual potential evaporation are likely to have an adverse affect on species living in these habitats, resulting in further bottlenecks and possibly local extinction. An IUCN assessment of P. nuptialis and P. bowkeri suggests that these two species are probably endangered. The issue surrounding the conservation of Prestonella species is that they are threatened by global climate change, which cannot be simply restricted or prevented, which makes dealing with the threat of climate change difficult. Assisted migration (MA) may be considered as a method to prevent possible future extinctions of Prestonella populations, but will only be considered as a last resort. The thermal tolerance (Arrhenius breaking temperature and flat-line temperature) of individual snails from three Prestonella populations (one forest population and two thicket populations) were assessed using infrared sensors that detected changes in heart rate with increasing temperature. The forest population had a significantly lower Arrhenius breaking temperature (ABT) and flat-line temperature (FLT) than the two thicket population (p<0.05). Our results do not show a correlation between upper thermal limits and maximum habitat temperatures or other climatic variables in Prestonella populations. Although no correlation is found between ABT and maximum habitat temperature, it is likely that the differences seen between these populations are due to local micro-climate adaptation. The climatic variables used in this experiment are coarse estimates from GIS data and do not reflect actual microhabitat conditions. Forest environments are less heat stressed than thicket environments due to the forest canopy which may explain the lower ABT and FLT of the forest population.
- Full Text:
- Date Issued: 2011
- Authors: Fearon, Janine Lee
- Date: 2011
- Subjects: Snails -- South Africa , Snails -- Conservation -- South Africa , Snails -- Variation -- South Africa , Biodiversity -- South Africa , Snails -- Genetics -- South Africa , Snails -- Habitat -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4191 , http://hdl.handle.net/10962/d1003760 , Snails -- South Africa , Snails -- Conservation -- South Africa , Snails -- Variation -- South Africa , Biodiversity -- South Africa , Snails -- Genetics -- South Africa , Snails -- Habitat -- South Africa
- Description: Prestonella bowkeri and Prestonella nuptialis are montane specialists endemic to the southern Great Escarpment of South Africa. Phylogeographic analyses of these species based on mitochondrial markers CO1 and 16S reveal extremely high levels of divergence between populations indicating a lack of gene flow between populations. This is not surprising, because P. nuptialis and P. bowkeri have limited dispersal capacity, low vagility, a highly fragmented distribution and are habitat specialists that are restricted to isolated mesic refugia associated with waterfalls and montane seepages. A relaxed Bayesian clock estimate suggests that populations diverged from one another during the mid-late Miocene (12.5-7 MYA) which coincides with the modern trends of seasonal aridity which began during the Miocene. This result should be viewed with caution because the rates used are at best imprecise estimates of mutation rates in snails. There is no clear dichotomy between the two species and P. bowkeri is paraphyletic with respect to P. nuptialis, as a consequence the taxonomy is unclear. Due to the high levels of sequence divergence between populations they may be considered as evolutionary significant units (ESU’s). An assessment of haplotype diversity (h) and nucleotide diversity (π) reveals that populations in the western part of the Great Escarpment are more genetically depauperate than populations in the east. Correlations between genetic diversity and climatic variables show that genetically depauperate populations are found in areas that have lower annual rainfall, less reliable rainfall and higher potential evaporation, all factors associated with a drier, less mesic environment that increases the chances of a population bottleneck. This indicates that a shift towards a more arid environment may be a driver of genetic erosion. Historical climate change may thus have affected the amount and distribution of genetic diversity across the Great Escarpment since the Miocene. This has serious future implications for the survival of Prestonella. With predicted increase in global temperatures, climate change in South Africa is likely to result in range contraction and an eastward range shift for many species in the drier central and western areas (Erasmus et al. 2002) and regions along the Great Escarpment are likely to become more arid. Prestonella populations found living on inselbergs along the Great Escarpment are already restricted to site specific watercourses and seepages. An increase in the periods between stream flow, and increasing rainfall variability and mean annual potential evaporation are likely to have an adverse affect on species living in these habitats, resulting in further bottlenecks and possibly local extinction. An IUCN assessment of P. nuptialis and P. bowkeri suggests that these two species are probably endangered. The issue surrounding the conservation of Prestonella species is that they are threatened by global climate change, which cannot be simply restricted or prevented, which makes dealing with the threat of climate change difficult. Assisted migration (MA) may be considered as a method to prevent possible future extinctions of Prestonella populations, but will only be considered as a last resort. The thermal tolerance (Arrhenius breaking temperature and flat-line temperature) of individual snails from three Prestonella populations (one forest population and two thicket populations) were assessed using infrared sensors that detected changes in heart rate with increasing temperature. The forest population had a significantly lower Arrhenius breaking temperature (ABT) and flat-line temperature (FLT) than the two thicket population (p<0.05). Our results do not show a correlation between upper thermal limits and maximum habitat temperatures or other climatic variables in Prestonella populations. Although no correlation is found between ABT and maximum habitat temperature, it is likely that the differences seen between these populations are due to local micro-climate adaptation. The climatic variables used in this experiment are coarse estimates from GIS data and do not reflect actual microhabitat conditions. Forest environments are less heat stressed than thicket environments due to the forest canopy which may explain the lower ABT and FLT of the forest population.
- Full Text:
- Date Issued: 2011
Molecular analysis of genetic diversity in dometicated pigeonpea (Cajanus cajan (L.) Millsp.) and wild relatives
- Authors: Kassa, Mulualem Tamiru
- Date: 2011
- Subjects: Pigeon pea -- Variation Cajanus -- Variation Pigeon pea -- Genetics Cajanus -- Genetics Biodiversity Pigeon pea -- Phylogeny Cajanus -- Phylogeny Plant hybridization Plant diversity Transgenic plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4204 , http://hdl.handle.net/10962/d1003773
- Description: Cajanus cajan (L.) Millsp. (Pigeonpea) belongs to the Leguminosae genus Cajanus which is composed of 34 species. Pigeonpea is the only cultivated member of the genus, while the remaining species are wild relatives belonging mainly to the secondary gene pool. DNA sequence data from the nuclear ITS region and the chloroplast trnL-F spacer were utilized to investigate the phylogenetic relationships between Cajanus and five other allied genera in the subtribe Cajaninae. This study revealed the non-monophyly of Cajanus and Rhynchosia and supported the monophyly of Eriosema and Flemingia, but more sampling ,especially from the large genera of Rhynchosia and Eriosema, is recommend to adequately test the hypothesis of generic monophyly. The phylogenetic relationships within the genus Cajanus resolved Cajanus scarabaeoides (L.) Thouars as the most basal species in the Cajanus clade. The study also utilized Single Nucleotide Polymorphism (SNP) markers derived from low copy orthologous genes and genotyped using the high throughput SNP-OPA Illumina golden gate assay. The aim was to understand phylogenetic and domestication history, genetic structure, patterns of genetic diversity, gene flow and historical hybridization between Cajanus cajan (pigeonpea) and wild relatives. The neighbor-joining tree resolved well-supported clusters, which reflect the distinctiveness of species and congruence with their geographical origin. It supported the ITS based phylogeny and resolved C. scarabaeoides as basal to the Cajanus clade. The phylogenetic signal and genetic signatures revealed insights into the domestication history of pigeonpea. Our results supported Cajanus cajanifolius as the presumed progenitor of pigeonpea and we speculate that for pigeonpea there was a single major domestication event in India. Genetic admixture and historical hybridization were evident between pigeonpea and wild relatives. Abundant allelic variation and genetic diversity was found in the wild relatives, with the exception of wild species from Australia, as compared to the domesticated pigeonpea. There was a reduction of about 75% in genetic polymorphism in domesticated pigeonpea as compared to the wild relatives, indicating a severe “domestication bottleneck” during pigeonpea domestication. We discovered SNP markers associated with disease resistance (NBS-LRR) loci. The SNPs were mined in a comparison of BAC-end sequences (BES) of C. cajan and amplicons of the wild species, C. scarabaeoides. A total of ~3000 SNPs were identified from 304 BES. These SNPs could potentially be used in constructing a genetic map and for marker assisted breeding.
- Full Text:
- Date Issued: 2011
- Authors: Kassa, Mulualem Tamiru
- Date: 2011
- Subjects: Pigeon pea -- Variation Cajanus -- Variation Pigeon pea -- Genetics Cajanus -- Genetics Biodiversity Pigeon pea -- Phylogeny Cajanus -- Phylogeny Plant hybridization Plant diversity Transgenic plants
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4204 , http://hdl.handle.net/10962/d1003773
- Description: Cajanus cajan (L.) Millsp. (Pigeonpea) belongs to the Leguminosae genus Cajanus which is composed of 34 species. Pigeonpea is the only cultivated member of the genus, while the remaining species are wild relatives belonging mainly to the secondary gene pool. DNA sequence data from the nuclear ITS region and the chloroplast trnL-F spacer were utilized to investigate the phylogenetic relationships between Cajanus and five other allied genera in the subtribe Cajaninae. This study revealed the non-monophyly of Cajanus and Rhynchosia and supported the monophyly of Eriosema and Flemingia, but more sampling ,especially from the large genera of Rhynchosia and Eriosema, is recommend to adequately test the hypothesis of generic monophyly. The phylogenetic relationships within the genus Cajanus resolved Cajanus scarabaeoides (L.) Thouars as the most basal species in the Cajanus clade. The study also utilized Single Nucleotide Polymorphism (SNP) markers derived from low copy orthologous genes and genotyped using the high throughput SNP-OPA Illumina golden gate assay. The aim was to understand phylogenetic and domestication history, genetic structure, patterns of genetic diversity, gene flow and historical hybridization between Cajanus cajan (pigeonpea) and wild relatives. The neighbor-joining tree resolved well-supported clusters, which reflect the distinctiveness of species and congruence with their geographical origin. It supported the ITS based phylogeny and resolved C. scarabaeoides as basal to the Cajanus clade. The phylogenetic signal and genetic signatures revealed insights into the domestication history of pigeonpea. Our results supported Cajanus cajanifolius as the presumed progenitor of pigeonpea and we speculate that for pigeonpea there was a single major domestication event in India. Genetic admixture and historical hybridization were evident between pigeonpea and wild relatives. Abundant allelic variation and genetic diversity was found in the wild relatives, with the exception of wild species from Australia, as compared to the domesticated pigeonpea. There was a reduction of about 75% in genetic polymorphism in domesticated pigeonpea as compared to the wild relatives, indicating a severe “domestication bottleneck” during pigeonpea domestication. We discovered SNP markers associated with disease resistance (NBS-LRR) loci. The SNPs were mined in a comparison of BAC-end sequences (BES) of C. cajan and amplicons of the wild species, C. scarabaeoides. A total of ~3000 SNPs were identified from 304 BES. These SNPs could potentially be used in constructing a genetic map and for marker assisted breeding.
- Full Text:
- Date Issued: 2011
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