Evaluation of pharmacological properties and anti-neoplastic potentials of pelargonium iquinans and opuntia stricta extracts using acute leukemia cell lines
- Authors: Izuegbuna, Ogochukwu Osita
- Date: 2018
- Subjects: Pelargoniums Medicinal plants
- Language: English
- Type: Thesis , Masters , M Sc
- Identifier: http://hdl.handle.net/10353/10372 , vital:35442
- Description: Leukaemia like other cancers is a public health issue, and coupled with the issue of drug resistance, toxicity and refractory disease novel treatments both as definitive and supportive care become necessary. The dissertation was designed to evaluate two medicinal plants (Opuntia stricta and Pelargonium inquinans) and their antineoplastic potentials. The phytochemicals analyses were done using standard methods with three different solvents (acetone, aqueous and ethanol). Vitamins analyses was also done using standard methods. The result revealed that vitamin A,C and E were present in the plants. Cell based anti-inflammatory activity were carried out using RAW264.7 macrophages cell, and COX-2 and NO expression as inflammatory markers. Cell based cytotoxicity and antineoplastic activity was investigated using RAW 264.7 macrophages cell, U937 and Jurkat cells. The results of the phytochemical analyses using GC/MS showed that phenols, flavonoids, flavonol, tannin, proanthocyanidin, saponin, alkaloids and phytate were present in these plants. In vitro antioxidant activity was carried out using nitric oxide (NO), alpha, alpha-diphenyl- β-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2), phosphomolybdenum activity, and iron reducing power (FRAP) techniques. The results of the antioxidant activity showed both plants had antioxidant properties that can be attributed to the phytochemical and vitamins contents. The results showed that both plants had anti-inflammatory properties which can also be attributed to their phytochemicals. The GC-MS analysis of the essential oils of both plants showed the presence of monoterpenes, sesquiterpenes, fatty acids, and esters. The results showed that O. stricta extracts have mild cytotoxicity, while P. inquinans have strong cytotoxicity. The cell cycle analysis showed Pelargonium inquinans extracts induced G0/G1 phase arrest and cleaved caspase-3. This study has shown the antineoplastic potential of P. inquinans which may be attributed to some phytochemicals in it such as carophyllene, humulene, beta myrcene and beta pinene.
- Full Text:
- Authors: Izuegbuna, Ogochukwu Osita
- Date: 2018
- Subjects: Pelargoniums Medicinal plants
- Language: English
- Type: Thesis , Masters , M Sc
- Identifier: http://hdl.handle.net/10353/10372 , vital:35442
- Description: Leukaemia like other cancers is a public health issue, and coupled with the issue of drug resistance, toxicity and refractory disease novel treatments both as definitive and supportive care become necessary. The dissertation was designed to evaluate two medicinal plants (Opuntia stricta and Pelargonium inquinans) and their antineoplastic potentials. The phytochemicals analyses were done using standard methods with three different solvents (acetone, aqueous and ethanol). Vitamins analyses was also done using standard methods. The result revealed that vitamin A,C and E were present in the plants. Cell based anti-inflammatory activity were carried out using RAW264.7 macrophages cell, and COX-2 and NO expression as inflammatory markers. Cell based cytotoxicity and antineoplastic activity was investigated using RAW 264.7 macrophages cell, U937 and Jurkat cells. The results of the phytochemical analyses using GC/MS showed that phenols, flavonoids, flavonol, tannin, proanthocyanidin, saponin, alkaloids and phytate were present in these plants. In vitro antioxidant activity was carried out using nitric oxide (NO), alpha, alpha-diphenyl- β-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2), phosphomolybdenum activity, and iron reducing power (FRAP) techniques. The results of the antioxidant activity showed both plants had antioxidant properties that can be attributed to the phytochemical and vitamins contents. The results showed that both plants had anti-inflammatory properties which can also be attributed to their phytochemicals. The GC-MS analysis of the essential oils of both plants showed the presence of monoterpenes, sesquiterpenes, fatty acids, and esters. The results showed that O. stricta extracts have mild cytotoxicity, while P. inquinans have strong cytotoxicity. The cell cycle analysis showed Pelargonium inquinans extracts induced G0/G1 phase arrest and cleaved caspase-3. This study has shown the antineoplastic potential of P. inquinans which may be attributed to some phytochemicals in it such as carophyllene, humulene, beta myrcene and beta pinene.
- Full Text:
Pollution potential of on-site dry sanitation systems associated with the Mzimvubu Water Project, Eastern Cape, South Africa
- Authors: Mamera, Matthew
- Date: 2018
- Subjects: Water resources development Sustainable development -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/6038 , vital:29485
- Description: Protecting the environment requires tools that can be used to monitor environmental conditions for proper sustainable management of water resources. The Mzimvubu River is the largest undeveloped river, in the poorest region of South Africa. The South African government announced plans to construct two large storage dams (Ntabelanga and Laleni) in the Tsitsa River, one of the largest tributaries to the Mzimvubu River. The dams and associated infrastructure is termed the Mzimvubu Water Project (MWP). In developing countries like South Africa, several households use pit latrines. Moreover, many people also rely upon untreated groundwater supplies for their drinking water. The shared utilization of both pit latrines and groundwater resources, can lead to human and ecological health risks from microbial and chemical pollutants. These threats can be accelerated due to a permanent rise in the water table associated with dam constructions. Four hillslope sites with pit latrines (MT1, MT2, MT3 and MT4) above 1st order tributaries to the Tsitsa River, in the vicinity of the proposed MWP were chosen for this study. The soils were described and classified at selected locations below the pit latrine and above the tributary. Undisturbed core samples were from representative soil horizons on which hydraulic properties were determined in the laboratory. Samples were also collected to determine the feacal coliform and other bacterial concentrations during a winter and summer period. Mechanistic modelling with Hydrus 2D for a 121 day period was conducted to determine the risk of pollution to surface and groundwater from the various sites. Apedal soils of the Clovelly form dominated upper slopes of MT1 and the lower slopes of MT2. A Clovelly form is hydropedologically termed a recharge soil, as morphological indicators of saturation are absent. Hydromorphic soil properties were observed in the sub-surface B horizons of a Tukulu form in MT1-3 and MT4. High clay contents and strong structure soils of the Sepane form were dominant in MT3. The soils of MT1 and MT2 are relatively sandy with slightly higher hydraulic conductivities compared to MT3 and MT4. In general higher measured faecal coliform concentrations were higher in winter as compared to summer seasons but in most sites both winter and summer seasons had counts < 1 CFU/g. The highest concentrations of total bacteria (9 x 106 CFU/ g soil) was observed in MT3-1. Other sites with potential pollution include MT3-3, with a count above 4 x 104 CFU /g soil in winter and MT1-1 with a count above 2.2 X 104 CFU /g soil in summer. In most of the sites, the E. coli bacteria detections were low (< 1 CFU/g) during both seasons. The highest value in summer of 1.8 x 102 CFU/g was observed in MT3-3 which increased from 1.2 x102 CFU/g in winter. As the soil water flow rate increase the mobility of bacteria increase as well. Hydrus simulations showed that surface water resources are threatened by pollution from MT3 and MT4 due to the prominence of lateral flow. Groundwater pollution is more likely to occur on MT1 and MT2 as the soils are freely drained. Future work should focus on the more direct measurements and modelling of the migration of bacterial pollutants from pit latrines to various water resources.
- Full Text:
- Authors: Mamera, Matthew
- Date: 2018
- Subjects: Water resources development Sustainable development -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/6038 , vital:29485
- Description: Protecting the environment requires tools that can be used to monitor environmental conditions for proper sustainable management of water resources. The Mzimvubu River is the largest undeveloped river, in the poorest region of South Africa. The South African government announced plans to construct two large storage dams (Ntabelanga and Laleni) in the Tsitsa River, one of the largest tributaries to the Mzimvubu River. The dams and associated infrastructure is termed the Mzimvubu Water Project (MWP). In developing countries like South Africa, several households use pit latrines. Moreover, many people also rely upon untreated groundwater supplies for their drinking water. The shared utilization of both pit latrines and groundwater resources, can lead to human and ecological health risks from microbial and chemical pollutants. These threats can be accelerated due to a permanent rise in the water table associated with dam constructions. Four hillslope sites with pit latrines (MT1, MT2, MT3 and MT4) above 1st order tributaries to the Tsitsa River, in the vicinity of the proposed MWP were chosen for this study. The soils were described and classified at selected locations below the pit latrine and above the tributary. Undisturbed core samples were from representative soil horizons on which hydraulic properties were determined in the laboratory. Samples were also collected to determine the feacal coliform and other bacterial concentrations during a winter and summer period. Mechanistic modelling with Hydrus 2D for a 121 day period was conducted to determine the risk of pollution to surface and groundwater from the various sites. Apedal soils of the Clovelly form dominated upper slopes of MT1 and the lower slopes of MT2. A Clovelly form is hydropedologically termed a recharge soil, as morphological indicators of saturation are absent. Hydromorphic soil properties were observed in the sub-surface B horizons of a Tukulu form in MT1-3 and MT4. High clay contents and strong structure soils of the Sepane form were dominant in MT3. The soils of MT1 and MT2 are relatively sandy with slightly higher hydraulic conductivities compared to MT3 and MT4. In general higher measured faecal coliform concentrations were higher in winter as compared to summer seasons but in most sites both winter and summer seasons had counts < 1 CFU/g. The highest concentrations of total bacteria (9 x 106 CFU/ g soil) was observed in MT3-1. Other sites with potential pollution include MT3-3, with a count above 4 x 104 CFU /g soil in winter and MT1-1 with a count above 2.2 X 104 CFU /g soil in summer. In most of the sites, the E. coli bacteria detections were low (< 1 CFU/g) during both seasons. The highest value in summer of 1.8 x 102 CFU/g was observed in MT3-3 which increased from 1.2 x102 CFU/g in winter. As the soil water flow rate increase the mobility of bacteria increase as well. Hydrus simulations showed that surface water resources are threatened by pollution from MT3 and MT4 due to the prominence of lateral flow. Groundwater pollution is more likely to occur on MT1 and MT2 as the soils are freely drained. Future work should focus on the more direct measurements and modelling of the migration of bacterial pollutants from pit latrines to various water resources.
- Full Text:
Development and validation of DNA barcoding for biodiversity assessment and conservation of red algae in the Algoa Bay region
- Authors: Mshiywa, Faith Masilive
- Date: 2017
- Subjects: Red algae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/9011 , vital:34182
- Description: DNA barcoding is a molecular technique that uses a short DNA sequence for species identification. It harnesses global community efforts to establish large-scale public reference libraries to allow species identification. Red algae are a complex group of species and are difficult to identify on morphological grounds only. Red algae have the highest production of halogenated compounds, and are used in the medical and pharmaceutical industries. This study therefore aimed at developing DNA barcodes, universal primers and a Kenton-on-Sea red algae species list, which will aid as a baseline for red algae biodiversity and conservation research. A potential DNA barcode region was designed from the rbcL gene, because rbcL is easy to amplify, easy to sequence and it can differentiate organisms to species level. Primers were designed from multiple red algae rbcL sequences and they were evaluated using bioinformatics tools. Red algae from the Algoa Bay region were screened with these primers and a 77% PCR amplification success was obtained. The PCR products were sequenced and 96% of the amplicons were successfully sequenced and resulted in a barcode sequence length above 700bp, which could be used to identify red algae species. Twenty-six (26) red algae species from Kenton-on-Sea, near Algoa Bay, were successfully identified using the Barcode sequence amplified from a primer set developed from the rbcL gene and were uploaded on the BOLD database for public access. This barcode is, therefore, suitable for use by the South African National Biodiversity Institute or other marine researchers, to identify South African red algae species and ensure the maintenance of their biodiversity and conservation.
- Full Text:
- Authors: Mshiywa, Faith Masilive
- Date: 2017
- Subjects: Red algae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/9011 , vital:34182
- Description: DNA barcoding is a molecular technique that uses a short DNA sequence for species identification. It harnesses global community efforts to establish large-scale public reference libraries to allow species identification. Red algae are a complex group of species and are difficult to identify on morphological grounds only. Red algae have the highest production of halogenated compounds, and are used in the medical and pharmaceutical industries. This study therefore aimed at developing DNA barcodes, universal primers and a Kenton-on-Sea red algae species list, which will aid as a baseline for red algae biodiversity and conservation research. A potential DNA barcode region was designed from the rbcL gene, because rbcL is easy to amplify, easy to sequence and it can differentiate organisms to species level. Primers were designed from multiple red algae rbcL sequences and they were evaluated using bioinformatics tools. Red algae from the Algoa Bay region were screened with these primers and a 77% PCR amplification success was obtained. The PCR products were sequenced and 96% of the amplicons were successfully sequenced and resulted in a barcode sequence length above 700bp, which could be used to identify red algae species. Twenty-six (26) red algae species from Kenton-on-Sea, near Algoa Bay, were successfully identified using the Barcode sequence amplified from a primer set developed from the rbcL gene and were uploaded on the BOLD database for public access. This barcode is, therefore, suitable for use by the South African National Biodiversity Institute or other marine researchers, to identify South African red algae species and ensure the maintenance of their biodiversity and conservation.
- Full Text:
Genetics, physiology, proteomics of quality protein maize inbred lines under drought and heat stress
- Authors: Pfunde, Cleopatra Nyaradzo
- Date: 2016
- Subjects: Corn Drought-tolerant plants Proteomics
- Language: English
- Type: Thesis , Masters/Doctoral , Crop Science
- Identifier: http://hdl.handle.net/10353/11696 , vital:39098
- Description: When plants perceive heat stress alone or in combination with drought, they activate several responses to synchronize development and molecular activities to ensure survival. In this study, a multidiscipline approach was adopted to investigate the plant genetics, proteomics and physiology for understanding the mechanisms underlying the adaptive response to combined drought and heat stress. The quality protein maize (QPM) inbred lines used had varied degrees of tolerance to both heat stress alone and in combination with drought. The overall objective of this study was to identify drought and heat tolerant germplasm that combine well under drought and heat stress for use in breeding programs to develop new QPM varieties. Initially, the genetic diversity among available inbred lines was estimated using two molecular markers, simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). The results indicated that moderate diversity exists among the QPM inbred lines which can be exploited to generate high performing hybrids. Cross combinations between QS26 and CIM2 and those between QS6 and CIM14 were selected as crosses likely to give substantial heterosis in the field because of the high genetic distances that were found between them using SNPs. Screening of inbred lines for drought and heat tolerance was done at the early germination and seedling stages. Differences in heat tolerance at both stages of growth were observed. Heat tolerant inbred lines at early germination stage had longer coleoptiles and radicles after recovery. The following inbred lines exhibited acquired thermo-tolerance, QS6, CIM7, QS17 and QS19, while CIM12, QS1, CIM21 and QS32 showed basal thermo-tolerance. At the early seedling stage inbred lines with significant differences (P< 0.001) in morpho-physiological traits were noted under heat stress alone and under combined drought and heat stress. However, no significant differences were noted for morpho-physiological measurements in the control treatment. Electron transport rate, sub-stomatal CO2, and stomatal conductance were significantly (P<0.001) reduced in response to combined drought and heat stress. Thus, drought coupled with heat stress resulted in stomatal closure, decreased photosynthetic rate, enhanced respiration and increased leaf temperature for susceptible inbred lines. The combined effects of drought and heat stress were more deleterious than the individual effect of heat stress alone as indicated by the reduction in dry weight. Based on the physiological parameters, biomass and tolerance indices, inbred lines QS22, CIM18 and QS6 showed tolerance to combined drought and heat stress, while CIM18, QS14 and QS30 exhibited thermo-tolerance. Inbred line CIM18 showed cross tolerance to combined drought and heat stress, and to heat stress alone. Further investigations into the comparative proteomic analysis of CIM18 (tolerant) with QS21 (susceptible) under combined drought and heat stress at seedling stage revealed that photosynthetic and stress related proteins were upregulated in CIM18. In addition, the abundance of heat shock proteins such as HSP70 and sHSP 17.8 kDa meant that CIM18 was better able to maintain cellular homeostasis and metabolism. The presence of metabolism related proteins meant that the energy needs of photosynthesis and other activities were met under drought and heat stress. Anti-oxidant enzymes such as glutathione s transferase were responsible for detoxification of reactive oxygen species induced by oxidative stress. These proteins mediated tolerance to combined stress in CIM18. The proteins that were micro-sequenced have previously been identified in individual drought and heat stress studies. The expression of these proteins under combined stress indicated that mechanisms from both stresses were pooled to confer tolerance in CIM18. Accordingly, the study proposes that CIM18 is a drought and heat tolerant QPM inbred line and is recommended for inclusion in breeding programs aimed at the development of drought and heat tolerant varieties. QPM has the potential to improve human nutrition in several rural households. The combining ability analysis of inbred lines revealed that GCA effects under combined drought and heat stress were significant for ears per plant, chlorophyll content, canopy temperature, anthesis silking interval, days to 50percent silking and ear length. Specific combining ability effects were significant for grain yield, days to 50percent anthesis, chlorophyll content and senescence. The study revealed that non-additive gene action was more important in determining grain yield under combined drought and heat stress. Inbred lines L2, L3, L7 and L3, L7 and L11 were good sources of genes for high grain yield under well-watered and combined drought and heat stress respectively. Cross combinations L3 x L13 (3.05 t/ha) and L5 x L9 (2.95 t/ha) were the best performing single cross hybrids for grain yield while cross combination L2 x L10 (1.33t/ha) was the least performing hybrid under combined stress conditions. Under well-watered conditions, the highest yielding crosses were between L2 x L13 (6.51t/ha) and L2 x L8 (6.46 t/ha). The inbred lines identified as sources of drought and heat tolerance and the high yielding cross combinations maybe useful for improving tolerance by generating hybrids or synthetic QPM maize varieties. The inbred lines which exhibited tolerance to both drought and heat stress were genetically diverse and were able to maintain photosynthesis and hence growth during stressful conditions. The ability of these genotypes to accumulate biomass or produce yield under stressful conditions highlights underlying mechanisms which need to be further exploited. While partial proteomic work was done in this study, a more comprehensive study looking into greater depth at the genes responsible for these mechanisms is paramount to fully utilize and/or transfer these genes to other desirable genotypes.
- Full Text:
Genetics, physiology, proteomics of quality protein maize inbred lines under drought and heat stress
- Authors: Pfunde, Cleopatra Nyaradzo
- Date: 2016
- Subjects: Corn Drought-tolerant plants Proteomics
- Language: English
- Type: Thesis , Masters/Doctoral , Crop Science
- Identifier: http://hdl.handle.net/10353/11696 , vital:39098
- Description: When plants perceive heat stress alone or in combination with drought, they activate several responses to synchronize development and molecular activities to ensure survival. In this study, a multidiscipline approach was adopted to investigate the plant genetics, proteomics and physiology for understanding the mechanisms underlying the adaptive response to combined drought and heat stress. The quality protein maize (QPM) inbred lines used had varied degrees of tolerance to both heat stress alone and in combination with drought. The overall objective of this study was to identify drought and heat tolerant germplasm that combine well under drought and heat stress for use in breeding programs to develop new QPM varieties. Initially, the genetic diversity among available inbred lines was estimated using two molecular markers, simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). The results indicated that moderate diversity exists among the QPM inbred lines which can be exploited to generate high performing hybrids. Cross combinations between QS26 and CIM2 and those between QS6 and CIM14 were selected as crosses likely to give substantial heterosis in the field because of the high genetic distances that were found between them using SNPs. Screening of inbred lines for drought and heat tolerance was done at the early germination and seedling stages. Differences in heat tolerance at both stages of growth were observed. Heat tolerant inbred lines at early germination stage had longer coleoptiles and radicles after recovery. The following inbred lines exhibited acquired thermo-tolerance, QS6, CIM7, QS17 and QS19, while CIM12, QS1, CIM21 and QS32 showed basal thermo-tolerance. At the early seedling stage inbred lines with significant differences (P< 0.001) in morpho-physiological traits were noted under heat stress alone and under combined drought and heat stress. However, no significant differences were noted for morpho-physiological measurements in the control treatment. Electron transport rate, sub-stomatal CO2, and stomatal conductance were significantly (P<0.001) reduced in response to combined drought and heat stress. Thus, drought coupled with heat stress resulted in stomatal closure, decreased photosynthetic rate, enhanced respiration and increased leaf temperature for susceptible inbred lines. The combined effects of drought and heat stress were more deleterious than the individual effect of heat stress alone as indicated by the reduction in dry weight. Based on the physiological parameters, biomass and tolerance indices, inbred lines QS22, CIM18 and QS6 showed tolerance to combined drought and heat stress, while CIM18, QS14 and QS30 exhibited thermo-tolerance. Inbred line CIM18 showed cross tolerance to combined drought and heat stress, and to heat stress alone. Further investigations into the comparative proteomic analysis of CIM18 (tolerant) with QS21 (susceptible) under combined drought and heat stress at seedling stage revealed that photosynthetic and stress related proteins were upregulated in CIM18. In addition, the abundance of heat shock proteins such as HSP70 and sHSP 17.8 kDa meant that CIM18 was better able to maintain cellular homeostasis and metabolism. The presence of metabolism related proteins meant that the energy needs of photosynthesis and other activities were met under drought and heat stress. Anti-oxidant enzymes such as glutathione s transferase were responsible for detoxification of reactive oxygen species induced by oxidative stress. These proteins mediated tolerance to combined stress in CIM18. The proteins that were micro-sequenced have previously been identified in individual drought and heat stress studies. The expression of these proteins under combined stress indicated that mechanisms from both stresses were pooled to confer tolerance in CIM18. Accordingly, the study proposes that CIM18 is a drought and heat tolerant QPM inbred line and is recommended for inclusion in breeding programs aimed at the development of drought and heat tolerant varieties. QPM has the potential to improve human nutrition in several rural households. The combining ability analysis of inbred lines revealed that GCA effects under combined drought and heat stress were significant for ears per plant, chlorophyll content, canopy temperature, anthesis silking interval, days to 50percent silking and ear length. Specific combining ability effects were significant for grain yield, days to 50percent anthesis, chlorophyll content and senescence. The study revealed that non-additive gene action was more important in determining grain yield under combined drought and heat stress. Inbred lines L2, L3, L7 and L3, L7 and L11 were good sources of genes for high grain yield under well-watered and combined drought and heat stress respectively. Cross combinations L3 x L13 (3.05 t/ha) and L5 x L9 (2.95 t/ha) were the best performing single cross hybrids for grain yield while cross combination L2 x L10 (1.33t/ha) was the least performing hybrid under combined stress conditions. Under well-watered conditions, the highest yielding crosses were between L2 x L13 (6.51t/ha) and L2 x L8 (6.46 t/ha). The inbred lines identified as sources of drought and heat tolerance and the high yielding cross combinations maybe useful for improving tolerance by generating hybrids or synthetic QPM maize varieties. The inbred lines which exhibited tolerance to both drought and heat stress were genetically diverse and were able to maintain photosynthesis and hence growth during stressful conditions. The ability of these genotypes to accumulate biomass or produce yield under stressful conditions highlights underlying mechanisms which need to be further exploited. While partial proteomic work was done in this study, a more comprehensive study looking into greater depth at the genes responsible for these mechanisms is paramount to fully utilize and/or transfer these genes to other desirable genotypes.
- Full Text:
A preliminary study on the effects of elevated CO2 on aphid resistance of Tugela Dn and the population dynamics of the Russian wheat aphid (Homoptera: Aphididae), Diuraphis noxia
- Authors: Mundondo, Daphine
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11402 , http://hdl.handle.net/10353/d1020244
- Description: Food security is of major importance due to the increasing world population with 8.9 billion people expected by 2050 (Cohen, 2003). Diuraphis noxia (RWA), have caused aggravating, massive losses to wheat farmers in many areas of the world. If unchecked, RWA are able to destroy plants resulting in major economic impacts (Botha, 2013). Due to ineffective use of other control methods, the Small Grains Institute in Bethlehem, South Africa, have therefore developed resistant cultivars to the known RWA subtypes over the past decades through intensive breeding programmes (Tolmay et al., 2006). Climate change has however become a major factor threatening food security especially with the observed increase in CO2 from less than 300 ppm in pre-industrial period to the current 385 ppm and is predicted to reach 550 ppm by 2050 (IPCC, 2007; Meehl et al., 2007). Elevated CO2 concentration may affect individual species of a community hence the need to understand the wheat-aphid interactions. In this study, population growth rates and virulence of RWA SA1 at ambient (385 ppm) and elevated (450 ppm) CO2 concentration were evaluated on two wheat cultivars: Tugela Dn (resistant) and Scheepers (susceptible). Fluorescence microscopy techniques using aniline blue were used to investigate feeding related damage caused by RWA SA1 through an examination of callose deposition at the two CO2 concentration. A two-dimensional gel electrophoresis method was developed in order to determine the effect of RWA SA1 on the wheat cultivars proteome at the two CO2 concentration. Differentially expressed proteins that were up or down regulated more than two fold were identified using PDQuestTM Basic 2D Gel analysis software. Populations of RWA SA1 increased significantly on the two wheat cultivars at both CO2 concentration. Although the population growth rate for RWA SA1 on both cultivars was generally exponential at all treatments, growth at elevated CO2 concentration was noticeably faster with populations increasing 3 fold in 14 days as compared to the 2 times at ambient CO2 concentration. Hence, both cultivars provided a better quality host for RWA SA1 at 450 ppm than 385 ppm. There was no significant difference between RWA SA1 population on Tugela Dn and on Scheepers at elevated CO2 concentration on day 14 after infestation which means there was a change in the resistance mechanism in Tugela Dn at this condition. Approximately 70% of the total leaf showed chlorosis by 21 days of aphid infestation for both cultivars although the susceptible cultivar was more vulnerable. There was low callose deposition in the controls (uninfested plants) but heavy callose in infested plants due to aphid feeding. A proteomics approach was used as a pilot study to investigate whether it would be possible to identify the changes in the resistance mechanism during aphid infestation under elevated CO2 levels. The major changes in the proteome of the control group (uninfested Tugela Dn at ambient versus elevated CO2 concentration) occurred in the early events (day 1-7) in the molecular weight range of approximately 25 kDa to 55 kDa are mainly within the basic to neutral pH range. This was suggested to be a result of mechanisms to adjust to the CO2 concentration. Elevated CO2 results in instant higher photosynthetic rates and C:N ratios as well as changes in expression levels of SA-dependant defense genes (Lindroth 1995; Hughes and Bazzaz, 2001; Sun et al., 2013). Because most of these changes are directly regulated by proteins, it is expected that the most differential protein expression will occur immediately after the atmospheric changes (early events) as was shown in the study. Infested plants under elevated and ambient conditions showed that the stress conditions gave rise to differentially regulated proteins within the wheat proteome. Most changes occurred elevated CO2 levels. It can be suggested that the changes were a result of differentially regulated plant defence proteins which fall in this range (25 kDa - 80 kDa) such as peroxidases, chitinases and β-1.3-glucanases as well as protein kinases, heat-shock proteins and photosynthetic proteins. These results indicate that there has been changes in the resistance due to elevated CO2 because of the evident changes in the proteome. If so, then the results will be similar to those documented by Louw (2007) where up-regulation was due to putative storage proteins, proteins involved in photosynthesis, heat shock proteins and defense proteins. Of course, the pI value and molecular mass of the proteins and the identification of the proteins in these spots, must be determined in future work to specifically identify whether these suggestions are authentic. However, Louw (2007) also reports that the susceptible Betta wheat cultivar, displayed a defence response similar to the HR although it was unable to up-regulate specific defensive proteins against RWA infestation but proteins for broad resistance. Although the changes in the proteins in infested Tugela Dn under elevated CO2 concentration were not accurately identified, the defense mechanism is similar to that portrayed by the susceptible Betta wheat cultivar which shows that the resistance mechanism had been overcome. Because this was a pilot study and preliminary results were obtained due to limited funding and time constraints, suggestions were made on how to further develop the method to obtain statistically significant results.
- Full Text:
- Authors: Mundondo, Daphine
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11402 , http://hdl.handle.net/10353/d1020244
- Description: Food security is of major importance due to the increasing world population with 8.9 billion people expected by 2050 (Cohen, 2003). Diuraphis noxia (RWA), have caused aggravating, massive losses to wheat farmers in many areas of the world. If unchecked, RWA are able to destroy plants resulting in major economic impacts (Botha, 2013). Due to ineffective use of other control methods, the Small Grains Institute in Bethlehem, South Africa, have therefore developed resistant cultivars to the known RWA subtypes over the past decades through intensive breeding programmes (Tolmay et al., 2006). Climate change has however become a major factor threatening food security especially with the observed increase in CO2 from less than 300 ppm in pre-industrial period to the current 385 ppm and is predicted to reach 550 ppm by 2050 (IPCC, 2007; Meehl et al., 2007). Elevated CO2 concentration may affect individual species of a community hence the need to understand the wheat-aphid interactions. In this study, population growth rates and virulence of RWA SA1 at ambient (385 ppm) and elevated (450 ppm) CO2 concentration were evaluated on two wheat cultivars: Tugela Dn (resistant) and Scheepers (susceptible). Fluorescence microscopy techniques using aniline blue were used to investigate feeding related damage caused by RWA SA1 through an examination of callose deposition at the two CO2 concentration. A two-dimensional gel electrophoresis method was developed in order to determine the effect of RWA SA1 on the wheat cultivars proteome at the two CO2 concentration. Differentially expressed proteins that were up or down regulated more than two fold were identified using PDQuestTM Basic 2D Gel analysis software. Populations of RWA SA1 increased significantly on the two wheat cultivars at both CO2 concentration. Although the population growth rate for RWA SA1 on both cultivars was generally exponential at all treatments, growth at elevated CO2 concentration was noticeably faster with populations increasing 3 fold in 14 days as compared to the 2 times at ambient CO2 concentration. Hence, both cultivars provided a better quality host for RWA SA1 at 450 ppm than 385 ppm. There was no significant difference between RWA SA1 population on Tugela Dn and on Scheepers at elevated CO2 concentration on day 14 after infestation which means there was a change in the resistance mechanism in Tugela Dn at this condition. Approximately 70% of the total leaf showed chlorosis by 21 days of aphid infestation for both cultivars although the susceptible cultivar was more vulnerable. There was low callose deposition in the controls (uninfested plants) but heavy callose in infested plants due to aphid feeding. A proteomics approach was used as a pilot study to investigate whether it would be possible to identify the changes in the resistance mechanism during aphid infestation under elevated CO2 levels. The major changes in the proteome of the control group (uninfested Tugela Dn at ambient versus elevated CO2 concentration) occurred in the early events (day 1-7) in the molecular weight range of approximately 25 kDa to 55 kDa are mainly within the basic to neutral pH range. This was suggested to be a result of mechanisms to adjust to the CO2 concentration. Elevated CO2 results in instant higher photosynthetic rates and C:N ratios as well as changes in expression levels of SA-dependant defense genes (Lindroth 1995; Hughes and Bazzaz, 2001; Sun et al., 2013). Because most of these changes are directly regulated by proteins, it is expected that the most differential protein expression will occur immediately after the atmospheric changes (early events) as was shown in the study. Infested plants under elevated and ambient conditions showed that the stress conditions gave rise to differentially regulated proteins within the wheat proteome. Most changes occurred elevated CO2 levels. It can be suggested that the changes were a result of differentially regulated plant defence proteins which fall in this range (25 kDa - 80 kDa) such as peroxidases, chitinases and β-1.3-glucanases as well as protein kinases, heat-shock proteins and photosynthetic proteins. These results indicate that there has been changes in the resistance due to elevated CO2 because of the evident changes in the proteome. If so, then the results will be similar to those documented by Louw (2007) where up-regulation was due to putative storage proteins, proteins involved in photosynthesis, heat shock proteins and defense proteins. Of course, the pI value and molecular mass of the proteins and the identification of the proteins in these spots, must be determined in future work to specifically identify whether these suggestions are authentic. However, Louw (2007) also reports that the susceptible Betta wheat cultivar, displayed a defence response similar to the HR although it was unable to up-regulate specific defensive proteins against RWA infestation but proteins for broad resistance. Although the changes in the proteins in infested Tugela Dn under elevated CO2 concentration were not accurately identified, the defense mechanism is similar to that portrayed by the susceptible Betta wheat cultivar which shows that the resistance mechanism had been overcome. Because this was a pilot study and preliminary results were obtained due to limited funding and time constraints, suggestions were made on how to further develop the method to obtain statistically significant results.
- Full Text:
Identification of agricultural and industrial pollutants in the Kat River, Eastern Cape and their effect on agricultural products found along the river banks
- Authors: Mutingwende, Nhamo
- Date: 2015
- Subjects: Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11291 , http://hdl.handle.net/10353/d1020242 , Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Description: There is growing concern that commonly used Pharmaceuticals and Personal Care Products (PPCPs) and pesticides are entering and contaminating drinking water supplies. The use of targeted quantitation of PPCP has been well established but there is an emerging trend to also screen for and identify unexpected environmental pollutants. Chemicals like pesticides hormones and antibiotics are especially of interest because of proven endocrine disrupting effects and a possible development of bacterial resistance. Powerful screening methods are required to detect and quantify the presence of these compounds in our environment. PPCP encompass a wide range of pollutants, including Endocrine Disrupting Compounds (EDC), pesticides, hormones, antibiotics, drugs of abuse, x-ray contrast agents and drinking water disinfection by-products to name a few. In order to properly assess the effects of these compounds on our environment, it is necessary to accurately monitor their presence. The diversity of chemical properties of these compounds makes method development challenging. LC/MS/MS is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range. The new AB SCIEX TripleTOF™5600 LC/MS/MS was used to profile environmental samples for unexpected pollutants, to identify and characterise the chemical composition and structure of the pollutants, and to quantify (based on intensity) the concentration in collected water samples. Liquid Chromatography coupled to tandem Mass Spectrometry (LCMS/ MS) is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range, such as pesticides, antibiotics, drugs of abuse, x-ray contrast agents, drinking water disinfection by-products etc. More recently there is a growing interest from environmental researchers to also screen for and identify non-targeted compounds in environmental samples, including metabolites and degradates, but also completely unexpected pollutants. The new AB SCIEX TripleTOF™5600 LC/MS/MS system is capable of performing highly sensitive and fast MS scanning experiments to search for unknown molecular ions while also performing selective and characteristic MS/MS scanning for further compound identification and, therefore, is the instrument of choice for this challenging task. General unknown screening workflows do not use a target analyte list and compound detection is not based on any prior knowledge, including retention times and information on possible molecular and fragment ions. Therefore, acquired chromatograms are very rich in information and can easily contain thousands of ions from both any compounds present in the sample as well as from the sample matrix itself. Thus, powerful software tools are needed to explore such data to identify the unexpected compound. Water samples were collected both upstream and downstream of two WWTPs (Seymour and Fort Beaufort) and were directly injected on the AB SCIEX TripleTOF™5600 LC/MS/MS after being filtered. 15 sample points along the Kat River, ranging from a point as close to the source as possible to a point just before it joins the Great Fish River were used. The samples collected from the source were used as the control in each of the experiments, the assumption being the closer you get to the source, the less contaminated the water would be for the analysis of pesticides. Points were selected where the Kat River crosses the R67 or on farms where the river was accessible using farm roads. Samples were collected from October 2013 to November 2014.The Peak view software and Analyst software were used in the analysis of PPCPs. The XIC Manager allows you to manage large lists of compounds and perform automatic extracted ion chromatogram (XIC) calculations and review results operations. The results were displayed in the chromatogram pane and the XIC table (see results). The results reported here in this thesis indicate that there is contamination in the Kat River water due to both pesticides and PPCPs. The results also indicate that the food products are also contaminated and hence both the Kat River agricultural produce and its water need to be closely monitored for both pesticide and PPCPs contaminants. Further studies to investigate the quantitative levels of pesticides and PPCPs in the Kat river water to determine if the concentration levels of the detected pesticides are below the reported Maximum Residues Limits will be explored in the future.
- Full Text:
- Authors: Mutingwende, Nhamo
- Date: 2015
- Subjects: Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11291 , http://hdl.handle.net/10353/d1020242 , Environmental toxicology , Rivers -- Environmental aspects -- South Africa , Water -- Pollution -- Toxicology -- South Africa
- Description: There is growing concern that commonly used Pharmaceuticals and Personal Care Products (PPCPs) and pesticides are entering and contaminating drinking water supplies. The use of targeted quantitation of PPCP has been well established but there is an emerging trend to also screen for and identify unexpected environmental pollutants. Chemicals like pesticides hormones and antibiotics are especially of interest because of proven endocrine disrupting effects and a possible development of bacterial resistance. Powerful screening methods are required to detect and quantify the presence of these compounds in our environment. PPCP encompass a wide range of pollutants, including Endocrine Disrupting Compounds (EDC), pesticides, hormones, antibiotics, drugs of abuse, x-ray contrast agents and drinking water disinfection by-products to name a few. In order to properly assess the effects of these compounds on our environment, it is necessary to accurately monitor their presence. The diversity of chemical properties of these compounds makes method development challenging. LC/MS/MS is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range. The new AB SCIEX TripleTOF™5600 LC/MS/MS was used to profile environmental samples for unexpected pollutants, to identify and characterise the chemical composition and structure of the pollutants, and to quantify (based on intensity) the concentration in collected water samples. Liquid Chromatography coupled to tandem Mass Spectrometry (LCMS/ MS) is able to analyse polar, semi-volatile, and thermally labile compounds covering a wide molecular weight range, such as pesticides, antibiotics, drugs of abuse, x-ray contrast agents, drinking water disinfection by-products etc. More recently there is a growing interest from environmental researchers to also screen for and identify non-targeted compounds in environmental samples, including metabolites and degradates, but also completely unexpected pollutants. The new AB SCIEX TripleTOF™5600 LC/MS/MS system is capable of performing highly sensitive and fast MS scanning experiments to search for unknown molecular ions while also performing selective and characteristic MS/MS scanning for further compound identification and, therefore, is the instrument of choice for this challenging task. General unknown screening workflows do not use a target analyte list and compound detection is not based on any prior knowledge, including retention times and information on possible molecular and fragment ions. Therefore, acquired chromatograms are very rich in information and can easily contain thousands of ions from both any compounds present in the sample as well as from the sample matrix itself. Thus, powerful software tools are needed to explore such data to identify the unexpected compound. Water samples were collected both upstream and downstream of two WWTPs (Seymour and Fort Beaufort) and were directly injected on the AB SCIEX TripleTOF™5600 LC/MS/MS after being filtered. 15 sample points along the Kat River, ranging from a point as close to the source as possible to a point just before it joins the Great Fish River were used. The samples collected from the source were used as the control in each of the experiments, the assumption being the closer you get to the source, the less contaminated the water would be for the analysis of pesticides. Points were selected where the Kat River crosses the R67 or on farms where the river was accessible using farm roads. Samples were collected from October 2013 to November 2014.The Peak view software and Analyst software were used in the analysis of PPCPs. The XIC Manager allows you to manage large lists of compounds and perform automatic extracted ion chromatogram (XIC) calculations and review results operations. The results were displayed in the chromatogram pane and the XIC table (see results). The results reported here in this thesis indicate that there is contamination in the Kat River water due to both pesticides and PPCPs. The results also indicate that the food products are also contaminated and hence both the Kat River agricultural produce and its water need to be closely monitored for both pesticide and PPCPs contaminants. Further studies to investigate the quantitative levels of pesticides and PPCPs in the Kat river water to determine if the concentration levels of the detected pesticides are below the reported Maximum Residues Limits will be explored in the future.
- Full Text:
Phytochemical analysis and antibacterial properties of aqueous and ethanol extracts of Brachylaena elliptica (Thurb.) dc. and Brachylaena ilicifolia (Lam.) Phill & Schweick
- Authors: Sagbo, Idowu Jonas
- Date: 2015
- Subjects: Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11297 , http://hdl.handle.net/10353/d1021289 , Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Description: Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.
- Full Text:
- Authors: Sagbo, Idowu Jonas
- Date: 2015
- Subjects: Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11297 , http://hdl.handle.net/10353/d1021289 , Medicinal plants , Traditional medicine , Herbs -- Therapeutic use
- Description: Resistance of human pathogenic bacterial strains results in selective pressure against known antibiotic. However, plant derived compounds that possess antibacterial potential are currently being investigated for treatment of wound infections in diabetic patients as they are inexpensive and non-toxic. Hence, this dissertation was designed to evaluate two medicinal plants (Brachylaena elliptica and Brachylaena ilicifolia) traditionally used in the treatment of various diseases such as diabetes, and its secondary complications in diabetic patients. The in vitro antioxidant activity of both plants were evaluated using DPPH (1, 1-diphenylhydrazl), ferric reducing power, ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid), NO (nitric oxide) and H2O2 (hydrogen peroxide) techniques. The antibacterial test and Minimum inhibitory concentration (MIC) was determined by agar dilution method against 5 bacteria strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogene, Proteus vulgaris and Proteus mirabilis) infecting wounds in diabetic patients using amoxicillin and ciprofloxacin as positive control. The phytochemical analyses were assessed using standard published methods. Identification of bioactive components in essential oils of both plants were assessed using GCMS. The aqueous and ethanol extracts of both plants were also evaluated to identify bioactive components using LC-MS. The results of the phytochemical analysis revealed the presence of phenols, tannins, flavanoids, flavanols, proanthocyanidins, saponins and alkaloids in both plants. Both plants indicated strong antioxidant activities which might be due to the presence of bioactive compounds. The aqueous and ethanol leaf extracts of both plants demonstrated appreciable broad spectrum activities against these wound pathogens with MIC ranging between 5 and 0.3 mg/ml. The GC-MS analysis of the essential oils of both plants revealed the presence of monoterpenes, oxygenated sesquiterpenes, phenolics and esters. The LC-MS analysis of the aqueous and ethanol leaf extracts of both plants showed that both plants are rich in alkaloids, terpenes, terpenoids, monoterpernoids, and flavanoids. Conclusively, this study has partially justified the ethnomedicinal use of B. elliptica and B.licifolia leaves for the treatment of various diseases, including diabetes and wound infections caused by bacteria in diabetic patients. These may be attributed to the presence of antioxidant compound such as phenols, flavanoids, saponins, tannins, alkaloids and other phytochemical compounds.
- Full Text:
Evaluation of the efficacy of Carpobrotus edulis (L.) bolus leaf as a traditional treatment for the management of HIV/AIDS
- Authors: Omoruyi, Beauty Etinosa
- Date: 2014
- Subjects: AIDS (Disease) -- Treatment -- South Africa -- Eastern Cape , Phytochemicals -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Carpobrotus
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: http://hdl.handle.net/10353/744 , vital:26493 , AIDS (Disease) -- Treatment -- South Africa -- Eastern Cape , Phytochemicals -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Carpobrotus
- Description: The human immunodeficiency virus (HIV) is one of the most common and dreaded diseases of the 21th century. Today, the disease is still spreading with increasing incidence. “Since the beginning of the epidemic, almost 75 million people have been infected with the HIV virus and about 36 million people have died of HIV. Globally, 35.3 million [32.2–38.8 million] people were living with HIV at the end of 2012” (http://www.who.int/gho/hiv/en/). Several studies have been conducted on herbs under a multitude of ethnobotanical grounds. The use of medicinal plants for the management of HIV has become a common practice especially, in the Eastern Cape Province of South Africa (Wilfred Otang Mbeng, 2013 PhD thesis, UFH). At the beginning of this programme, an ethnomedicinal survey of plants used for the management of HIV infection was carried out in targeted areas of the Province and information on the names of plants, the parts and the methods of preparation were collected. The survey revealed that 18 species representing 12 families were found to be commonly used for the management of HIV, as well as other opportunistic diseases such as tuberculosis, diabetes mellitus, sores, high blood pressure, etc. Carpobrotus edulis was selected for this research because it was the most frequently used in the Province. The foliar micro morphological contents of the plant, its phytochemical and antioxidant activity, in vitro antimicrobial activity, inhibitory effect against HIV-1 protease and reverse transcriptase, mechanisms of action and cytotoxicity were investigated. In terms of the foliar micro morphological contents in plants, an electron microscopy scanning (SEM) was completed. Investigation revealed that both glandular tricomes and calcium oxalate crystals (CaOX) were observed. Consequently, it is hypothesized that the bioactive therapeutic compounds secreted by C. edulis may be produced in the glandular trichomes. An investigation of phytochemical content of the plant extracts (C. edulis) was completed using four solvent extracts (hexane, acetone, ethanol and water). Results of the phytochemical analysis showed that proanthocyanidins (86.9 ± 0.005%) where highest in the aqueous extract with phenolics at 55.7 ± 0.404% in acetone extract, tannin at 48.9 ± 0.28% in ethanol extract, while the hexane extract had the highest levels of flavonoids (0.12 ± 0.05%) and flavonols (0.12 ± 0.05%). Antioxidant studies of the various extracts revealed that aqueous and ethanol extracts were found to be the best solvents for antioxidant activity in C. edulis leaves. GC-MS analysis of the essential oil from C. edulis leaves revealed that the essential oil contained at least 28 compounds. These included, in order of abundance: Oxygenated monoterpenes (36.61%); fatty acids esters (19.25%); oxygenated diterpenes (19.24%); monoterpenes (10.6%); sesquiterpenes (3.58%); and diterpenes (1.43%). Similarly, a GC-MS analysis of the crude hexane, acetone and ethanol extracts from C. edulis leaves identified a total of 59 compounds. Of the 59 compounds, 12 major phyto-metabolites that are active against infectious diseases were identified. To comfirm the potential use of C. edulis to treat infectious disease, antifungal activity of the crude essential oil extract and the four solvent extracts were tested against Candida albicans, Candida krusei, Candida glabrata, Candida rugosa and Cryptococcus neoformans strains. The essential oil extract was found to be the most active against all the fungal strains tested and performed better than the four extracts used various solvents used to extract (hexane, acetone, ethanol and water) the C. edulis leaves were tested for antibacterial and anti HIV-1 reverse transcriptase (RT) activity. The results indicated that both gram-positive and gram-negative isolates were inhibited by the extracts (hexane, acetone and ethanol) but antimicrobial activity was observed for the water extract. The lowest minimum inhibitory concentration values were obtained for the ethanol extract, followed by acetone and hexane extracts. No inhibition of HIV-1 reverse transcriptase was observed for any of the leaf extracts, even up to concentrations of 16 mg/ml. The potential inhibitory activities of the various solvent extracts against HIV-1 protease were evaluated at four different concentrations (16, 1.6, 0.16 and 0.016 mg/ml). Results indicated that the water extract showed almost 100% inhibition of HIV-1 protease activity, with an IC50 of 0.86 mg/ml leaf extract. Other solvent extracts (hexane, acetone and ethanol) however, did not show any inhibition activity above that observed for the DMSO control. The metabolic components in the water extract were subjected to LC-MS/MS analysis, which identified at least 91 compounds present in the water extract. Further studies involving the molecular modelling need to be carried out to confirm the inhibitory potential of these compounds. The cytotoxicity of the water extract of C. edulis leaves was also screened using human Chang liver cells at concentrations ranging bewteen 0.005 mg/ml and 1 mg/ml. Results indicated that the water extracts were not toxic. In conclusion the results from this study support the use of water extracts of C. edulis leaves by traditional healers to treat HIV infections and have identified possible mechanisms of action of the water extracts of C. edulis.
- Full Text:
- Authors: Omoruyi, Beauty Etinosa
- Date: 2014
- Subjects: AIDS (Disease) -- Treatment -- South Africa -- Eastern Cape , Phytochemicals -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Carpobrotus
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: http://hdl.handle.net/10353/744 , vital:26493 , AIDS (Disease) -- Treatment -- South Africa -- Eastern Cape , Phytochemicals -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Carpobrotus
- Description: The human immunodeficiency virus (HIV) is one of the most common and dreaded diseases of the 21th century. Today, the disease is still spreading with increasing incidence. “Since the beginning of the epidemic, almost 75 million people have been infected with the HIV virus and about 36 million people have died of HIV. Globally, 35.3 million [32.2–38.8 million] people were living with HIV at the end of 2012” (http://www.who.int/gho/hiv/en/). Several studies have been conducted on herbs under a multitude of ethnobotanical grounds. The use of medicinal plants for the management of HIV has become a common practice especially, in the Eastern Cape Province of South Africa (Wilfred Otang Mbeng, 2013 PhD thesis, UFH). At the beginning of this programme, an ethnomedicinal survey of plants used for the management of HIV infection was carried out in targeted areas of the Province and information on the names of plants, the parts and the methods of preparation were collected. The survey revealed that 18 species representing 12 families were found to be commonly used for the management of HIV, as well as other opportunistic diseases such as tuberculosis, diabetes mellitus, sores, high blood pressure, etc. Carpobrotus edulis was selected for this research because it was the most frequently used in the Province. The foliar micro morphological contents of the plant, its phytochemical and antioxidant activity, in vitro antimicrobial activity, inhibitory effect against HIV-1 protease and reverse transcriptase, mechanisms of action and cytotoxicity were investigated. In terms of the foliar micro morphological contents in plants, an electron microscopy scanning (SEM) was completed. Investigation revealed that both glandular tricomes and calcium oxalate crystals (CaOX) were observed. Consequently, it is hypothesized that the bioactive therapeutic compounds secreted by C. edulis may be produced in the glandular trichomes. An investigation of phytochemical content of the plant extracts (C. edulis) was completed using four solvent extracts (hexane, acetone, ethanol and water). Results of the phytochemical analysis showed that proanthocyanidins (86.9 ± 0.005%) where highest in the aqueous extract with phenolics at 55.7 ± 0.404% in acetone extract, tannin at 48.9 ± 0.28% in ethanol extract, while the hexane extract had the highest levels of flavonoids (0.12 ± 0.05%) and flavonols (0.12 ± 0.05%). Antioxidant studies of the various extracts revealed that aqueous and ethanol extracts were found to be the best solvents for antioxidant activity in C. edulis leaves. GC-MS analysis of the essential oil from C. edulis leaves revealed that the essential oil contained at least 28 compounds. These included, in order of abundance: Oxygenated monoterpenes (36.61%); fatty acids esters (19.25%); oxygenated diterpenes (19.24%); monoterpenes (10.6%); sesquiterpenes (3.58%); and diterpenes (1.43%). Similarly, a GC-MS analysis of the crude hexane, acetone and ethanol extracts from C. edulis leaves identified a total of 59 compounds. Of the 59 compounds, 12 major phyto-metabolites that are active against infectious diseases were identified. To comfirm the potential use of C. edulis to treat infectious disease, antifungal activity of the crude essential oil extract and the four solvent extracts were tested against Candida albicans, Candida krusei, Candida glabrata, Candida rugosa and Cryptococcus neoformans strains. The essential oil extract was found to be the most active against all the fungal strains tested and performed better than the four extracts used various solvents used to extract (hexane, acetone, ethanol and water) the C. edulis leaves were tested for antibacterial and anti HIV-1 reverse transcriptase (RT) activity. The results indicated that both gram-positive and gram-negative isolates were inhibited by the extracts (hexane, acetone and ethanol) but antimicrobial activity was observed for the water extract. The lowest minimum inhibitory concentration values were obtained for the ethanol extract, followed by acetone and hexane extracts. No inhibition of HIV-1 reverse transcriptase was observed for any of the leaf extracts, even up to concentrations of 16 mg/ml. The potential inhibitory activities of the various solvent extracts against HIV-1 protease were evaluated at four different concentrations (16, 1.6, 0.16 and 0.016 mg/ml). Results indicated that the water extract showed almost 100% inhibition of HIV-1 protease activity, with an IC50 of 0.86 mg/ml leaf extract. Other solvent extracts (hexane, acetone and ethanol) however, did not show any inhibition activity above that observed for the DMSO control. The metabolic components in the water extract were subjected to LC-MS/MS analysis, which identified at least 91 compounds present in the water extract. Further studies involving the molecular modelling need to be carried out to confirm the inhibitory potential of these compounds. The cytotoxicity of the water extract of C. edulis leaves was also screened using human Chang liver cells at concentrations ranging bewteen 0.005 mg/ml and 1 mg/ml. Results indicated that the water extracts were not toxic. In conclusion the results from this study support the use of water extracts of C. edulis leaves by traditional healers to treat HIV infections and have identified possible mechanisms of action of the water extracts of C. edulis.
- Full Text:
Isolation, purification and kinetic characterization of prolyl endopeptidase from Titicum aestivum
- Authors: Abrahams, Adriam Mark
- Date: 2013
- Subjects: Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11269 , http://hdl.handle.net/10353/d1004356 , Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Description: PEP activity has been described in several locations and has mostly been linked to a variety of neurological disorders such as schizophrenia, amnaesia, depression as well as other disease states such as anorexia nervosa, bulimia nervosa and blood pressure regulation. The enzyme has also been previously isolated from a variety of archae, microorganisms and several eukaryotic species but no prolyl endopeptidases have been isolated from plants. Plants have high levels of proline and glutamine rich peptides in seeds. We therefore hypothesize plants must express PEPs during germination. Bioinformatics tools were used to identify known PEPs and putative plant PEPs. A global sequence alignment of putative plant PEPs and other known PEPs indicated that the active site amino acids Ser, His and Asp are conserved in putative plant PEP sequences. Furthermore, putative plant PEPs showed similar secondary structures to known PEPs and when a rice PEP was modelled onto porcine brain PEP structure, a high degree of similarity was found. Germination studies of wheat seed showed an increase of PEP activity over time with maximum PEP activity reached after 4 days of germination, which remained at this level until 9 days of germination, implying a function for PEP in plant seed germination. Wheat PEP was purified using ion exchange and gel filtration chromatography with a final yield of less than 1 percent and a relative purity (only 2 bands detected by SDS-PAGE). The purified wheat PEP had a molecular weight of approximately 55kDa, substrate specificity for chymotrypsin-like substrates (N-Suc-Ala-Ala-Pro-Phe-pNa, Km value of 0.58 mM, Kcat of 29.37 s–1; Kcat /Km 50813.14s–1 M–1); a pH optimum of 7.9; temperature optima of 37oC and a high sensitivity to temperature as indicated by loss of activity at temperatures above 40oC. Inhibition studies using E64, Leupeptin and PMSF confirmed that the wheat PEP is from the Serine protease family and is most likely a trypsin-like protease.
- Full Text:
- Authors: Abrahams, Adriam Mark
- Date: 2013
- Subjects: Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11269 , http://hdl.handle.net/10353/d1004356 , Endopeptidases , Wheat , Chemical kinetics , Pharmacokinetics
- Description: PEP activity has been described in several locations and has mostly been linked to a variety of neurological disorders such as schizophrenia, amnaesia, depression as well as other disease states such as anorexia nervosa, bulimia nervosa and blood pressure regulation. The enzyme has also been previously isolated from a variety of archae, microorganisms and several eukaryotic species but no prolyl endopeptidases have been isolated from plants. Plants have high levels of proline and glutamine rich peptides in seeds. We therefore hypothesize plants must express PEPs during germination. Bioinformatics tools were used to identify known PEPs and putative plant PEPs. A global sequence alignment of putative plant PEPs and other known PEPs indicated that the active site amino acids Ser, His and Asp are conserved in putative plant PEP sequences. Furthermore, putative plant PEPs showed similar secondary structures to known PEPs and when a rice PEP was modelled onto porcine brain PEP structure, a high degree of similarity was found. Germination studies of wheat seed showed an increase of PEP activity over time with maximum PEP activity reached after 4 days of germination, which remained at this level until 9 days of germination, implying a function for PEP in plant seed germination. Wheat PEP was purified using ion exchange and gel filtration chromatography with a final yield of less than 1 percent and a relative purity (only 2 bands detected by SDS-PAGE). The purified wheat PEP had a molecular weight of approximately 55kDa, substrate specificity for chymotrypsin-like substrates (N-Suc-Ala-Ala-Pro-Phe-pNa, Km value of 0.58 mM, Kcat of 29.37 s–1; Kcat /Km 50813.14s–1 M–1); a pH optimum of 7.9; temperature optima of 37oC and a high sensitivity to temperature as indicated by loss of activity at temperatures above 40oC. Inhibition studies using E64, Leupeptin and PMSF confirmed that the wheat PEP is from the Serine protease family and is most likely a trypsin-like protease.
- Full Text:
Evaluation of the possible application of cowpea genotypes in the farming systems of the Eastern Cape Province, South Africa
- Authors: Adeyemi, Samson Adebowale
- Date: 2012
- Subjects: Cowpea -- South Africa -- Eastern Cape , Cowpea , Plant diversity , Cowpea -- Genetics , DNA fingerprinting of plants
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11274 , http://hdl.handle.net/10353/d1007539 , Cowpea -- South Africa -- Eastern Cape , Cowpea , Plant diversity , Cowpea -- Genetics , DNA fingerprinting of plants
- Description: Characterization studies on the genetic diversity among cultivated cowpea (Vigna unguiculata (L.) varieties are valuable tools to optimize the use of available genetic resources by farmers, local communities, researchers and breeders. Eight cowpea (Vigna unguiculata (L.) genotypes ( Vegetable cowpea, Ivory grey, Okhalweni, Fahari, Fahari dark, 97K-1069-8, IT93K-73h, and 129-3) were subjected to molecular, morphological and agronomical characterization. DNA amplification fingerprinting markers were used to evaluate the genetic diversity among the eight genotypes. Nine random arbitrary primers were used to screen the eight genotypes to assess their ability to reveal polymorphisms in cowpea, and seven of them were selected for use in characterizing the total sample. A total of 43 bands were generated which are all polymorphic. On the average, the primers generated a total of 6.1 polymorphic bands. The resulting data-matrix included 43 analysed bands with a total of 344 characters. Neighbour joining analysis was used to generate the dendrogram, clustering the genotypes into two groups at an agglomerate coefficient of 0.30 irrespective of their geographical origins. The results also showed the presence of significant differences in morphological and quality traits among the genotypes. Fahari yielded the highest concentration of crude protein (46.51 mg/mg dry leaf) while Vegetable cowpea yielded the lowest (24.41 mg/mg dry leaf). The influence of manure was also found to be effective by increasing the crude protein content of the genotypes as shown by Fahari dark with an average of 53.53 mg/mg dry leaf as opposed to 39.85 mg/mg dry leaf without manure application. Although some small clusters grouped accessions of the same growth habits, a general lack of agreement between clustering and morphological features was observed. It can therefore be concluded that the significant differences between the molecular genetic analysis using DAF-PCR markers, morphologic characters and yield traits can be important tools to identify and discriminates the different cowpea genotypes.
- Full Text:
- Authors: Adeyemi, Samson Adebowale
- Date: 2012
- Subjects: Cowpea -- South Africa -- Eastern Cape , Cowpea , Plant diversity , Cowpea -- Genetics , DNA fingerprinting of plants
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11274 , http://hdl.handle.net/10353/d1007539 , Cowpea -- South Africa -- Eastern Cape , Cowpea , Plant diversity , Cowpea -- Genetics , DNA fingerprinting of plants
- Description: Characterization studies on the genetic diversity among cultivated cowpea (Vigna unguiculata (L.) varieties are valuable tools to optimize the use of available genetic resources by farmers, local communities, researchers and breeders. Eight cowpea (Vigna unguiculata (L.) genotypes ( Vegetable cowpea, Ivory grey, Okhalweni, Fahari, Fahari dark, 97K-1069-8, IT93K-73h, and 129-3) were subjected to molecular, morphological and agronomical characterization. DNA amplification fingerprinting markers were used to evaluate the genetic diversity among the eight genotypes. Nine random arbitrary primers were used to screen the eight genotypes to assess their ability to reveal polymorphisms in cowpea, and seven of them were selected for use in characterizing the total sample. A total of 43 bands were generated which are all polymorphic. On the average, the primers generated a total of 6.1 polymorphic bands. The resulting data-matrix included 43 analysed bands with a total of 344 characters. Neighbour joining analysis was used to generate the dendrogram, clustering the genotypes into two groups at an agglomerate coefficient of 0.30 irrespective of their geographical origins. The results also showed the presence of significant differences in morphological and quality traits among the genotypes. Fahari yielded the highest concentration of crude protein (46.51 mg/mg dry leaf) while Vegetable cowpea yielded the lowest (24.41 mg/mg dry leaf). The influence of manure was also found to be effective by increasing the crude protein content of the genotypes as shown by Fahari dark with an average of 53.53 mg/mg dry leaf as opposed to 39.85 mg/mg dry leaf without manure application. Although some small clusters grouped accessions of the same growth habits, a general lack of agreement between clustering and morphological features was observed. It can therefore be concluded that the significant differences between the molecular genetic analysis using DAF-PCR markers, morphologic characters and yield traits can be important tools to identify and discriminates the different cowpea genotypes.
- Full Text:
Inhibitory potential of honey on the enzymatic activity of Helicobacter pylori urease
- Authors: Matongo, Fredrick
- Date: 2012
- Subjects: Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11253 , http://hdl.handle.net/10353/431 , Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Description: Urease of Helicobacter pylori is an important virulence factor implicated in the pathogenesis of many clinical conditions, such as chronic gastritis, peptic ulceration, and gastric cancer. Many urease inhibitors have been discovered, like phosphorodiamidates, hydroxamic acid derivatives, and imidazoles. Despite good activities at the enzyme level and excellent kinetic properties most of them have not been used as therapeutic agents in vivo because of their side effects, toxicity and instability. This has led to much attention to focus on exploring the novel urease inhibitory activities of natural products because of their low toxicity and good bioavailability. Honey, a natural product has been used in folk medicine due to its antitumor, antioxidant, antimicrobial and anti-inflammatory properties. The aims of this study were to isolate, characterise, purify urease produced by H. pylori and investigate the inhibitory effects of solvent honey extracts on its enzymatic activity. Urease was found to be both surface-associated and cytoplasmic. Maximum cytoplasmic urease activity was found to occur after 72 hr whereas maximum extracellular urease activities were found to occur after 96 hr. Characterization of the crude cytoplasmic urease revealed optimal activity at a pH of 7.5 and temperature of 40°C. The kinetic parameters Vmax and Km were 45.32 U ml-1 and 61.11 mM respectively.The honey extracts inhibited the activity of the crude urease in a concentration dependent manner. The Lineweaver-Burk plots indicated a non-competitive type of inhibition against H. pylori urease. The two honey extracts gave promising inhibitory activities against urease of H. pylori. Thus the results of this study delineates that inhibition of urease can ease development in therapeutic and preventative approaches based on the enzymatic activity of this Helicobacter protein.
- Full Text:
- Authors: Matongo, Fredrick
- Date: 2012
- Subjects: Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11253 , http://hdl.handle.net/10353/431 , Honey , Helicobacter pylori infections , Enzyme inhibitors , Traditional medicine , Antifungal agents
- Description: Urease of Helicobacter pylori is an important virulence factor implicated in the pathogenesis of many clinical conditions, such as chronic gastritis, peptic ulceration, and gastric cancer. Many urease inhibitors have been discovered, like phosphorodiamidates, hydroxamic acid derivatives, and imidazoles. Despite good activities at the enzyme level and excellent kinetic properties most of them have not been used as therapeutic agents in vivo because of their side effects, toxicity and instability. This has led to much attention to focus on exploring the novel urease inhibitory activities of natural products because of their low toxicity and good bioavailability. Honey, a natural product has been used in folk medicine due to its antitumor, antioxidant, antimicrobial and anti-inflammatory properties. The aims of this study were to isolate, characterise, purify urease produced by H. pylori and investigate the inhibitory effects of solvent honey extracts on its enzymatic activity. Urease was found to be both surface-associated and cytoplasmic. Maximum cytoplasmic urease activity was found to occur after 72 hr whereas maximum extracellular urease activities were found to occur after 96 hr. Characterization of the crude cytoplasmic urease revealed optimal activity at a pH of 7.5 and temperature of 40°C. The kinetic parameters Vmax and Km were 45.32 U ml-1 and 61.11 mM respectively.The honey extracts inhibited the activity of the crude urease in a concentration dependent manner. The Lineweaver-Burk plots indicated a non-competitive type of inhibition against H. pylori urease. The two honey extracts gave promising inhibitory activities against urease of H. pylori. Thus the results of this study delineates that inhibition of urease can ease development in therapeutic and preventative approaches based on the enzymatic activity of this Helicobacter protein.
- Full Text:
Parent characterization of quality protein maize (Zea mays L.) and combining ability for tolerance to drought stress
- Authors: Pfunde, Cleopatra Nyaradzo
- Date: 2012
- Subjects: Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Language: English
- Type: Thesis , Masters , MSc Agric (Crop Science)
- Identifier: vital:11869 , http://hdl.handle.net/10353/d1007536 , Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Description: Quality protein maize (QPM) has enhanced levels of two essential amino acids, lysine and tryptophan compared to normal maize. This makes QPM an important cereal crop in communities where maize is a staple crop. The main abiotic factor to QPM production is drought stress. Little information is available on the effect of drought stress on QPM. Therefore, the objectives of this study were to: (i) conduct diversity analysis of QPM inbred lines using morpho-agronomic and simple sequence repeat markers, (ii) screen available QPM inbred lines and F1 progeny for tolerance to seedling drought stress, (iii) determine the combining ability and type of gene action of QPM inbred lines for tolerance to seedling drought stress, grain yield and endosperm modification. The study was conducted in South Africa, at the University of Fort Hare. Morphological characterisation of 21 inbred lines was done using quantitative and qualitative traits. A randomised complete block design with three replicates was used for characterizing the inbred lines in the field. Genstat statistical software, version 12 (Genstat ®, 2009) was used for analysis of variance (ANOVA) and descriptive statistics. Analysis of variance was performed on all quantitative data for morphological traits. Data for qualitative traits was tabulated in their nominal classes. Traits that contributed most to the variation were days to anthesis, days to silking, anthesis-silking interval, plant height, number of kernel rows, ear length and grain yield. Cluster analysis grouped the inbred lines into three main clusters. The first cluster was characterised by tall and average yielding lines, while the second cluster showed the least anthesis-silking interval, and had the highest yield. Cluster three consisted of lines that were early maturing, but were the least yielding. Genetic distances between maize inbred lines were quantified by using 27 simple sequence repeat markers. The genetic distances between genotypes was computed using Roger’s (1972) genetic distances. Cluster analysis was then carried out using the neighbour-joining tree method using Power Marker software version 3.25. A dendrogram generated from the genetic study of the inbred lines revealed three groups that concurred with expectations based upon pedigree data. These groups were not identical to the groups generated using morpho-agronomic characterisation. Twenty one QPM inbred lines were crossed using a North Carolina design II mating scheme. These were divided into seven sets, each with three inbred lines. The three inbred lines in one set were used as females and crossed with three inbred lines in another set consisting of males. Each inbred line was used as a female in one set, and as a male in a second set. Sixty three hybrids (7 sets x 9 hybrids) were formed and evaluated in October 2011, using a 6x8 alpha-lattice incomplete block design with three replicates under glasshouse and optimum field conditions. A randomised complete block design with three replicates was used for the 21 parental inbred lines. Traits recorded for the glasshouse study were, canopy temperature, chlorophyll content, leaf roll, stem diameter, plant height, leaf number, leaf area, fresh and dry root and shoot weights. Data for the various traits for each environment, 25 percent (stress treatment) and 75 percent (non-stress) of field capacity, were subjected to analysis of variance using the unbalanced treatment design in Genstat statistical package Edition 12. Where varietal differences were found, means were separated using Tukey’s test. Genetic analyses for grain yield and agronomic traits were performed using a fixed effects model in JMP 10 following Residual Maximum Likelihood procedure (REML). From the results, inbred lines that were not previously classified into heterotic groups and drought tolerance categories were classified based on their total dry weight performance and drought susceptibility index. Inbred lines L18, L9, L8, L6 and L3, in order of their drought tolerance index were the best performers under greenhouse conditions and could be recommended for breeding new varieties that are tolerant to seedling drought stress. Evaluation of maize seedlings tolerant to drought stress under glasshouse conditions revealed that cross combination L18 x L11 was drought tolerant, while cross L20 x L7 was susceptible. Total dry weight was used as the major criteria for classifying F1 maize seedlings as being resistant or susceptible. General combining ability effects accounted for 67.43 percent of the genetic variation for total dry weight, while specific combining ability effects contributed 37.57 percent. This indicated that additive gene effects were more important than non-additive gene action in controlling this trait. In the field study (non-drought), the experimental design was a 6x8 alpha lattice incomplete block design with three replicates. On an adjacent field a randomised complete block design with three replicates was used to evaluate the parental inbred lines. The following variables were recorded: plant height, ear height, ears per plant, endosperm modification, days to silking and days to anthesis, anthesis-silking interval, number of kernels per row, number of rows per ear and grain yield. General analyses for the incomplete lattice block design and randomised complete block design for hybrid and inbred data respectively were performed using JMP 10 statistical software. Means were separated using the Tukey's test. Genetic analyses of data for grain yield and agronomic traits were conducted using a fixed effects model using REML in JMP 10. The importance of both GCA (51 percent) and SCA (49 percent) was observed for grain yield. A preponderance of GCA existed for ear height, days to anthesis, anthesis-silking interval, ears per plant and number of kernels per row, indicating that predominantly, additive gene effects controlled hybrid performance under optimum field conditions. The highest heritability was observed for days to silking (48.27 percent) suggesting that yield could be improved through selection for this trait. Under field conditions, variation in time to maturity was observed. This implies that these inbred lines can be recommended for utilisation in different agro-ecologies. Early maturing lines such as L18 can be used to introduce earliness in local cultivars, while early maturing single crosses such as L18 x L2, L5 x L9, L3 x L4 and L2 x L21 could be recommended for maize growers in drought prone areas such as the former Ciskei. Single crosses L18xL11, L16xL18, L8xL21 and L9xL6 had good tolerance to seedling drought stress. On the other hand, single crosses L18xL11 and L11xL13 had high grain yield and good endosperm modification. All these single crosses could be recommended for commercial production after evaluation across locations in the Eastern Cape Province. Alternatively they can be crossed with other superior inbreds to generate three or four way hybrids, which could then be evaluated for potential use by farmers in the Eastern Cape.
- Full Text:
- Authors: Pfunde, Cleopatra Nyaradzo
- Date: 2012
- Subjects: Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Language: English
- Type: Thesis , Masters , MSc Agric (Crop Science)
- Identifier: vital:11869 , http://hdl.handle.net/10353/d1007536 , Corn -- Quality , Corn as food , Corn -- Effect of stress on , Corn -- Effect of drought on , Cluster analysis , Crops -- Effect of drought on , Corn -- Drought tolerance , Corn -- Breeding , Crops -- Drought tolerance
- Description: Quality protein maize (QPM) has enhanced levels of two essential amino acids, lysine and tryptophan compared to normal maize. This makes QPM an important cereal crop in communities where maize is a staple crop. The main abiotic factor to QPM production is drought stress. Little information is available on the effect of drought stress on QPM. Therefore, the objectives of this study were to: (i) conduct diversity analysis of QPM inbred lines using morpho-agronomic and simple sequence repeat markers, (ii) screen available QPM inbred lines and F1 progeny for tolerance to seedling drought stress, (iii) determine the combining ability and type of gene action of QPM inbred lines for tolerance to seedling drought stress, grain yield and endosperm modification. The study was conducted in South Africa, at the University of Fort Hare. Morphological characterisation of 21 inbred lines was done using quantitative and qualitative traits. A randomised complete block design with three replicates was used for characterizing the inbred lines in the field. Genstat statistical software, version 12 (Genstat ®, 2009) was used for analysis of variance (ANOVA) and descriptive statistics. Analysis of variance was performed on all quantitative data for morphological traits. Data for qualitative traits was tabulated in their nominal classes. Traits that contributed most to the variation were days to anthesis, days to silking, anthesis-silking interval, plant height, number of kernel rows, ear length and grain yield. Cluster analysis grouped the inbred lines into three main clusters. The first cluster was characterised by tall and average yielding lines, while the second cluster showed the least anthesis-silking interval, and had the highest yield. Cluster three consisted of lines that were early maturing, but were the least yielding. Genetic distances between maize inbred lines were quantified by using 27 simple sequence repeat markers. The genetic distances between genotypes was computed using Roger’s (1972) genetic distances. Cluster analysis was then carried out using the neighbour-joining tree method using Power Marker software version 3.25. A dendrogram generated from the genetic study of the inbred lines revealed three groups that concurred with expectations based upon pedigree data. These groups were not identical to the groups generated using morpho-agronomic characterisation. Twenty one QPM inbred lines were crossed using a North Carolina design II mating scheme. These were divided into seven sets, each with three inbred lines. The three inbred lines in one set were used as females and crossed with three inbred lines in another set consisting of males. Each inbred line was used as a female in one set, and as a male in a second set. Sixty three hybrids (7 sets x 9 hybrids) were formed and evaluated in October 2011, using a 6x8 alpha-lattice incomplete block design with three replicates under glasshouse and optimum field conditions. A randomised complete block design with three replicates was used for the 21 parental inbred lines. Traits recorded for the glasshouse study were, canopy temperature, chlorophyll content, leaf roll, stem diameter, plant height, leaf number, leaf area, fresh and dry root and shoot weights. Data for the various traits for each environment, 25 percent (stress treatment) and 75 percent (non-stress) of field capacity, were subjected to analysis of variance using the unbalanced treatment design in Genstat statistical package Edition 12. Where varietal differences were found, means were separated using Tukey’s test. Genetic analyses for grain yield and agronomic traits were performed using a fixed effects model in JMP 10 following Residual Maximum Likelihood procedure (REML). From the results, inbred lines that were not previously classified into heterotic groups and drought tolerance categories were classified based on their total dry weight performance and drought susceptibility index. Inbred lines L18, L9, L8, L6 and L3, in order of their drought tolerance index were the best performers under greenhouse conditions and could be recommended for breeding new varieties that are tolerant to seedling drought stress. Evaluation of maize seedlings tolerant to drought stress under glasshouse conditions revealed that cross combination L18 x L11 was drought tolerant, while cross L20 x L7 was susceptible. Total dry weight was used as the major criteria for classifying F1 maize seedlings as being resistant or susceptible. General combining ability effects accounted for 67.43 percent of the genetic variation for total dry weight, while specific combining ability effects contributed 37.57 percent. This indicated that additive gene effects were more important than non-additive gene action in controlling this trait. In the field study (non-drought), the experimental design was a 6x8 alpha lattice incomplete block design with three replicates. On an adjacent field a randomised complete block design with three replicates was used to evaluate the parental inbred lines. The following variables were recorded: plant height, ear height, ears per plant, endosperm modification, days to silking and days to anthesis, anthesis-silking interval, number of kernels per row, number of rows per ear and grain yield. General analyses for the incomplete lattice block design and randomised complete block design for hybrid and inbred data respectively were performed using JMP 10 statistical software. Means were separated using the Tukey's test. Genetic analyses of data for grain yield and agronomic traits were conducted using a fixed effects model using REML in JMP 10. The importance of both GCA (51 percent) and SCA (49 percent) was observed for grain yield. A preponderance of GCA existed for ear height, days to anthesis, anthesis-silking interval, ears per plant and number of kernels per row, indicating that predominantly, additive gene effects controlled hybrid performance under optimum field conditions. The highest heritability was observed for days to silking (48.27 percent) suggesting that yield could be improved through selection for this trait. Under field conditions, variation in time to maturity was observed. This implies that these inbred lines can be recommended for utilisation in different agro-ecologies. Early maturing lines such as L18 can be used to introduce earliness in local cultivars, while early maturing single crosses such as L18 x L2, L5 x L9, L3 x L4 and L2 x L21 could be recommended for maize growers in drought prone areas such as the former Ciskei. Single crosses L18xL11, L16xL18, L8xL21 and L9xL6 had good tolerance to seedling drought stress. On the other hand, single crosses L18xL11 and L11xL13 had high grain yield and good endosperm modification. All these single crosses could be recommended for commercial production after evaluation across locations in the Eastern Cape Province. Alternatively they can be crossed with other superior inbreds to generate three or four way hybrids, which could then be evaluated for potential use by farmers in the Eastern Cape.
- Full Text:
Differential expression and regulation of sucrose transporters in rice (Orzya sativa L, cv Nipponbare) during environmental stress conditions
- Authors: Ibraheem, Omodele
- Date: 2011
- Subjects: Crops -- Effect of stress on , Plant molecular genetics , Gene expression , Sucrose , Rice
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11249 , http://hdl.handle.net/10353/330 , Crops -- Effect of stress on , Plant molecular genetics , Gene expression , Sucrose , Rice
- Description: Plant productivity is greatly affected by environmental stresses such as drought, salinity and insect herbivory. Plants respond and adapt to these stresses by exhibiting physiological as well as biochemical changes at the cellular and molecular levels in order to survive. Expression of a variety of genes which encode numerous membrane transporters have been demonstrated to be induced by these stresses in a variety of plants. The nutritional status of plants is controlled by these transporters, which are regulated by the transcription of the corresponding genes. In spite of these adverse stress effects on agricultural yield, only a few studies have focused on gene transcriptional and translational regulation of membrane transporters during environmental stress situations. Rice, like other plants, contains a number of sucrose transporters encoded by a family of genes. However, detailed knowledge of their roles, localization and regulation during environmental stress conditions is lacking. Bioinformatic tools were used to identify putative cis-acting regulatory elements that may be involved in the regulation of rice and Arabidopsis thaliana sucrose transporters. The possible cis-acting regulatory elements were predicted by scanning genomic sequences 1.5 kbp upstream of the sucrose transporter genes translational start sites, using Plant CARE, PLACE and Genomatix Matinspector professional data bases. Several cis-acting regulatory elements that are associated with plant development, plant hormonal regulation and stress response were identified, and were present in varying frequencies within the 1.5 kbp of 5′ regulatory region. The putative cis-acting regulatory elements that possibly are involved in the expression and regulation of sucrose transporter gene families in rice and Arabidopsis thaliana during cellular development or environmental stress conditions were identified as: A-box, RY, CAT, Pyrimidine-box, Sucrose-box, ABRE, ARF, ERE, GARE, Me-JA, ARE, DRE, GA-motif, GATA, GT-1, MYC, MYB, W-box, and I-box. Expression analysis was used to elucidate the role of rice (Oryza sativa L. cv Nipponbare) sucrose transporter (OsSUT) genes during drought and salinity treatments of three week old rice plants ( at four leaf stage) over a 10 days. Among the five rice OsSUT genes identified, only OsSUT2 was observed to be progressively up-regulated during drought and salinity treatments, while OsSUT1, OsSUT4 and OsSUT5 were expressed at low levels, and OsSUT3 showed no detectable transcript expression. Sucrose transport will be essential to meet the cellular energy demands and also for osmoprotectant activities during drought and salinity stresses. It therefore indicates that OsSUT2 which facilitates transport of sucrose from photosynthetic cells will be III essential for rice plants to cope with drought and salinity stresses, and cultivars with a higher OsSUT2 expression should be able to tolerate these environmental stresses better. The role of OsSUT in assimilate transport during rusty plum aphids (Hysteroneura setariae; Thomas) infestation on the leaves of three week old rice (Orzya sativa L. cv Nipponbare) cultivar plants, over a time-course of 1 to 10 days of treatments, was also examined by combination of gene expression and β-glucuronidase (GUS) reporter gene analysis. Real Time PCR analysis of the five OsSUT genes revealed that the expression of OsSUT1 was progressively up-regulated during the course of aphid infestation. OsSUT2 and OsSUT4 expression were comparatively low in both the control and treated plants. OsSUT5 showed no clear difference in transcript expression in both control and treated plants, while no detectable transcript expression of OsSUT3 could be found. The up-regulation of OsSUT1 gene was verified at protein level by western blot analysis in both the control and treated plants. OsSUT1 protein expression was found to increase with time during aphid infestation. A similar trend was noticeable in the control plants, however at a lower expression level. These demonstrate that the cellular expression of OsSUT1is regulated by both developmental and environmental factors. OsSUT1-promoter:::GUS reporter gene expression was observed within the vascular parenchyma and/or companion cells associated with phloem sieve elements of the large and small bundles in the phloem tissues of the flag leaf blade regions where feeding aphids were confined, which progressively increased with time of infestation. It is suggested that OsSUT1 may primarily play an essential role in phloem transport of assimilate to wounded tissues from adjacent health tissues or may be involved in the retrieval of assimilate back into the phloem to minimize loss caused by the infestation. Some OsSUT1-promoter:::GUS expression was also found in the metaxylem at 10 days after infestation, which could signify a recovery system in which sucrose lost into the xylem as a result of aphids feeding are retrieved back into the phloem through the vascular parenchyma. This was supported by the exposure of cut ends of matured OsSUT1-promoter:::GUS rice plant leaf to 2% sucrose solution. OsSUT1-promoter:::GUS expression was observed within the protoxylem, xylem and phloem parenchyma tissues. This indicates that sucrose translocating within the xylem tissues are retrieved into the phloem via the OsSUT1 localized within the parenchyma tissues. In conclusion, the differential expression and regulation of rice (Orzya sativa L. cv Nipponbare) sucrose transporters as reported here suggest that OsSUT2 and OsSUT1 were constitutively expressed compared to other rice sucrose transporters during drought and salinity, and rusty plum aphids (Hysteroneura setariae; Thomas) infestation stresses respectively. Thus, the expression and regulation of the sucrose transporters could be related to the physiological and nutritional requirements of the cells during plant developmental or environmental stress state that allows their differential expression.
- Full Text:
- Authors: Ibraheem, Omodele
- Date: 2011
- Subjects: Crops -- Effect of stress on , Plant molecular genetics , Gene expression , Sucrose , Rice
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11249 , http://hdl.handle.net/10353/330 , Crops -- Effect of stress on , Plant molecular genetics , Gene expression , Sucrose , Rice
- Description: Plant productivity is greatly affected by environmental stresses such as drought, salinity and insect herbivory. Plants respond and adapt to these stresses by exhibiting physiological as well as biochemical changes at the cellular and molecular levels in order to survive. Expression of a variety of genes which encode numerous membrane transporters have been demonstrated to be induced by these stresses in a variety of plants. The nutritional status of plants is controlled by these transporters, which are regulated by the transcription of the corresponding genes. In spite of these adverse stress effects on agricultural yield, only a few studies have focused on gene transcriptional and translational regulation of membrane transporters during environmental stress situations. Rice, like other plants, contains a number of sucrose transporters encoded by a family of genes. However, detailed knowledge of their roles, localization and regulation during environmental stress conditions is lacking. Bioinformatic tools were used to identify putative cis-acting regulatory elements that may be involved in the regulation of rice and Arabidopsis thaliana sucrose transporters. The possible cis-acting regulatory elements were predicted by scanning genomic sequences 1.5 kbp upstream of the sucrose transporter genes translational start sites, using Plant CARE, PLACE and Genomatix Matinspector professional data bases. Several cis-acting regulatory elements that are associated with plant development, plant hormonal regulation and stress response were identified, and were present in varying frequencies within the 1.5 kbp of 5′ regulatory region. The putative cis-acting regulatory elements that possibly are involved in the expression and regulation of sucrose transporter gene families in rice and Arabidopsis thaliana during cellular development or environmental stress conditions were identified as: A-box, RY, CAT, Pyrimidine-box, Sucrose-box, ABRE, ARF, ERE, GARE, Me-JA, ARE, DRE, GA-motif, GATA, GT-1, MYC, MYB, W-box, and I-box. Expression analysis was used to elucidate the role of rice (Oryza sativa L. cv Nipponbare) sucrose transporter (OsSUT) genes during drought and salinity treatments of three week old rice plants ( at four leaf stage) over a 10 days. Among the five rice OsSUT genes identified, only OsSUT2 was observed to be progressively up-regulated during drought and salinity treatments, while OsSUT1, OsSUT4 and OsSUT5 were expressed at low levels, and OsSUT3 showed no detectable transcript expression. Sucrose transport will be essential to meet the cellular energy demands and also for osmoprotectant activities during drought and salinity stresses. It therefore indicates that OsSUT2 which facilitates transport of sucrose from photosynthetic cells will be III essential for rice plants to cope with drought and salinity stresses, and cultivars with a higher OsSUT2 expression should be able to tolerate these environmental stresses better. The role of OsSUT in assimilate transport during rusty plum aphids (Hysteroneura setariae; Thomas) infestation on the leaves of three week old rice (Orzya sativa L. cv Nipponbare) cultivar plants, over a time-course of 1 to 10 days of treatments, was also examined by combination of gene expression and β-glucuronidase (GUS) reporter gene analysis. Real Time PCR analysis of the five OsSUT genes revealed that the expression of OsSUT1 was progressively up-regulated during the course of aphid infestation. OsSUT2 and OsSUT4 expression were comparatively low in both the control and treated plants. OsSUT5 showed no clear difference in transcript expression in both control and treated plants, while no detectable transcript expression of OsSUT3 could be found. The up-regulation of OsSUT1 gene was verified at protein level by western blot analysis in both the control and treated plants. OsSUT1 protein expression was found to increase with time during aphid infestation. A similar trend was noticeable in the control plants, however at a lower expression level. These demonstrate that the cellular expression of OsSUT1is regulated by both developmental and environmental factors. OsSUT1-promoter:::GUS reporter gene expression was observed within the vascular parenchyma and/or companion cells associated with phloem sieve elements of the large and small bundles in the phloem tissues of the flag leaf blade regions where feeding aphids were confined, which progressively increased with time of infestation. It is suggested that OsSUT1 may primarily play an essential role in phloem transport of assimilate to wounded tissues from adjacent health tissues or may be involved in the retrieval of assimilate back into the phloem to minimize loss caused by the infestation. Some OsSUT1-promoter:::GUS expression was also found in the metaxylem at 10 days after infestation, which could signify a recovery system in which sucrose lost into the xylem as a result of aphids feeding are retrieved back into the phloem through the vascular parenchyma. This was supported by the exposure of cut ends of matured OsSUT1-promoter:::GUS rice plant leaf to 2% sucrose solution. OsSUT1-promoter:::GUS expression was observed within the protoxylem, xylem and phloem parenchyma tissues. This indicates that sucrose translocating within the xylem tissues are retrieved into the phloem via the OsSUT1 localized within the parenchyma tissues. In conclusion, the differential expression and regulation of rice (Orzya sativa L. cv Nipponbare) sucrose transporters as reported here suggest that OsSUT2 and OsSUT1 were constitutively expressed compared to other rice sucrose transporters during drought and salinity, and rusty plum aphids (Hysteroneura setariae; Thomas) infestation stresses respectively. Thus, the expression and regulation of the sucrose transporters could be related to the physiological and nutritional requirements of the cells during plant developmental or environmental stress state that allows their differential expression.
- Full Text:
Investigation of antidiabetic properties, mechanisms of action and toxicology of Strychnos Henningsii (GILG) bark
- Authors: Oyewole, Oyedemi Sunday
- Date: 2011
- Subjects: Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11254 , http://hdl.handle.net/10353/d1001070 , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Description: The apparent reversal of trend from modern drugs to herbal medicine is partly due to the fact that synthetic drugs have always shown adverse reactions and other undesirable side effects. Hence, the use of medicinal plants for the treatment of diseases such as diabetes is very common especially in the rural areas. Majority of these plants are used based on the experience and indigenous knowledge without identification of the therapeutic agents. There is enormous wealth of medicinal plants in the world yet many of them have not been discovered or studied scientifically to substantiate their ethno-medicinal usages. Ethnobotanical study has been the method often used to search for locally important plant species for the discovery of crude drugs with low side effects. An ethnobotanical survey was conducted on the medicinal plants commonly used for the management of diabetes mellitus in Nkonkobe Municipality, Eastern Cape of South Africa. Information was obtained through structured questionnaire administered to traditional healers and herbalists in the region. The study revealed 15 species of plants belonging to 13 families. Strychnos henningsii and Leonotis leonorus among others were repeatedly mentioned by the traditional healers as the two mostly used plants for the management of diabetes mellitus. The infusion and decoction of the roots, leaves and barks of these plants are the methods of preparation. The antioxidant potential of aqueous bark extract of S. henningsii was investigated both in vivo and in vitro using spectroscopic method. The antioxidant activity of the extract against hydrogen peroxide (H2O2), 2,2′-azinobis[3-ethylbenzothiazoline6-sulfonic acid] diammonium salt (ABTS), as well as reducing power was concentration dependent. The extract exhibited lower and average scavenging activities against 1,1diphenyl2picrylhydrazyl (DPPH) and nitric oxide (NO) radicals with IC50 value of 0.739 and 0.49 mg/ml respectively. The administration of the plant extract at 250, 500 and 1000 mg/kg significantly increased the activities of the antioxidant enzymes in the hepatotoxic rats induced with carbon tetrachloride. On the other hand, the stem bark extract had lower effect on lipid peroxidation level except at the dose of 250 mg/kg. The effect of oral administration of S. henningsii extract was evaluated in normal Wistar rats for 28 days. The observed result indicated non- toxic effect of sub-acute administration of plant extract to the animals except at certain doses. This is because, there was no apparent damage to some haematological and biochemical parameters used in assessing organ specific toxicity. However, the alterations observed on platelet, white blood cells and its differentials imply parameter and dose selective toxicity when repeatedly consumed on daily basis at the doses investigated. This study also investigated the antidiabetic activities of the extract at the doses of 125, 250 and 500 mg/kg body weight in diabetic rats induced with streptozotocin -nicotinamide for 15 days. The extract appreciably (P <0.05) reduced the blood glucose level, feed and water intake while the best result was obtained at 250 mg/kg. Similarly, the level of triacylglycerol at the three doses investigated was significantly decreased. In addition, the glucose tolerance was reduced to near normal level after 90 min at certain doses. The clinical significance of the extract on some biochemical and haematological parameters lessen both hepatic and renal damages. Anaemic condition in diabetic animals was also improved after plant extract administration. However, no significant effect was observed in white blood cells and some of its differentials. The extract demonstrated strong glucose utilization in 3T3-L1 cells with a response of 278.63 percent of the control at 12.5μg/ml while that of Chang liver cells was 103.54 percent. The cytotoxicity result revealed non toxic effects of the extract to both cell lines. Treatment of 3T3 L1 cells with the extract did not reduce lipid accumulation. The extract inhibited the activity of α- glucosidase and α- amylase in a concentration dependent manner with IC50 values of 38 μg/ml and 60.9 μg/ml respectively. The percentage protein antiglycation of S. henningsii was 18.4, 38.2 and 61.2 perceent for 0.25, 0.5 and 1 mg/ml respectively while aminoguanidine a known inhibitor of protein glycation was 87.2 percent at 1 mg/ml. The FRAP assay values of the extract was 357.05 μmol Fe (II)/g. The findings from this study support the folkloric usage of this plant for the management of diabetes mellitus in the region.
- Full Text:
- Authors: Oyewole, Oyedemi Sunday
- Date: 2011
- Subjects: Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Language: English
- Type: Thesis , Doctoral , PhD (Biochemistry)
- Identifier: vital:11254 , http://hdl.handle.net/10353/d1001070 , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape , Diabetics -- Alternative treatment -- South Africa , Loganiaceae , Lamiaceae , Diabetes , Proteins
- Description: The apparent reversal of trend from modern drugs to herbal medicine is partly due to the fact that synthetic drugs have always shown adverse reactions and other undesirable side effects. Hence, the use of medicinal plants for the treatment of diseases such as diabetes is very common especially in the rural areas. Majority of these plants are used based on the experience and indigenous knowledge without identification of the therapeutic agents. There is enormous wealth of medicinal plants in the world yet many of them have not been discovered or studied scientifically to substantiate their ethno-medicinal usages. Ethnobotanical study has been the method often used to search for locally important plant species for the discovery of crude drugs with low side effects. An ethnobotanical survey was conducted on the medicinal plants commonly used for the management of diabetes mellitus in Nkonkobe Municipality, Eastern Cape of South Africa. Information was obtained through structured questionnaire administered to traditional healers and herbalists in the region. The study revealed 15 species of plants belonging to 13 families. Strychnos henningsii and Leonotis leonorus among others were repeatedly mentioned by the traditional healers as the two mostly used plants for the management of diabetes mellitus. The infusion and decoction of the roots, leaves and barks of these plants are the methods of preparation. The antioxidant potential of aqueous bark extract of S. henningsii was investigated both in vivo and in vitro using spectroscopic method. The antioxidant activity of the extract against hydrogen peroxide (H2O2), 2,2′-azinobis[3-ethylbenzothiazoline6-sulfonic acid] diammonium salt (ABTS), as well as reducing power was concentration dependent. The extract exhibited lower and average scavenging activities against 1,1diphenyl2picrylhydrazyl (DPPH) and nitric oxide (NO) radicals with IC50 value of 0.739 and 0.49 mg/ml respectively. The administration of the plant extract at 250, 500 and 1000 mg/kg significantly increased the activities of the antioxidant enzymes in the hepatotoxic rats induced with carbon tetrachloride. On the other hand, the stem bark extract had lower effect on lipid peroxidation level except at the dose of 250 mg/kg. The effect of oral administration of S. henningsii extract was evaluated in normal Wistar rats for 28 days. The observed result indicated non- toxic effect of sub-acute administration of plant extract to the animals except at certain doses. This is because, there was no apparent damage to some haematological and biochemical parameters used in assessing organ specific toxicity. However, the alterations observed on platelet, white blood cells and its differentials imply parameter and dose selective toxicity when repeatedly consumed on daily basis at the doses investigated. This study also investigated the antidiabetic activities of the extract at the doses of 125, 250 and 500 mg/kg body weight in diabetic rats induced with streptozotocin -nicotinamide for 15 days. The extract appreciably (P <0.05) reduced the blood glucose level, feed and water intake while the best result was obtained at 250 mg/kg. Similarly, the level of triacylglycerol at the three doses investigated was significantly decreased. In addition, the glucose tolerance was reduced to near normal level after 90 min at certain doses. The clinical significance of the extract on some biochemical and haematological parameters lessen both hepatic and renal damages. Anaemic condition in diabetic animals was also improved after plant extract administration. However, no significant effect was observed in white blood cells and some of its differentials. The extract demonstrated strong glucose utilization in 3T3-L1 cells with a response of 278.63 percent of the control at 12.5μg/ml while that of Chang liver cells was 103.54 percent. The cytotoxicity result revealed non toxic effects of the extract to both cell lines. Treatment of 3T3 L1 cells with the extract did not reduce lipid accumulation. The extract inhibited the activity of α- glucosidase and α- amylase in a concentration dependent manner with IC50 values of 38 μg/ml and 60.9 μg/ml respectively. The percentage protein antiglycation of S. henningsii was 18.4, 38.2 and 61.2 perceent for 0.25, 0.5 and 1 mg/ml respectively while aminoguanidine a known inhibitor of protein glycation was 87.2 percent at 1 mg/ml. The FRAP assay values of the extract was 357.05 μmol Fe (II)/g. The findings from this study support the folkloric usage of this plant for the management of diabetes mellitus in the region.
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Evolutionary development and functional role of plant natriuretic peptide (PNP)-B
- Authors: Hove, Runyararo Memory
- Date: 2009
- Subjects: Plant hormones , Peptides , Plant gene expression , Peptide hormones , Peptides -- Separation
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11251 , http://hdl.handle.net/10353/155 , Plant hormones , Peptides , Plant gene expression , Peptide hormones , Peptides -- Separation
- Description: Plant natriuretic peptides (PNP) are novel peptides which, like in vertebrates, have been shown to have a function associated with water and salt homeostasis. Two PNP-encoding genes have been identified and isolated from Arabidopsis thaliana, namely; AtPNP-A and AtPNP-B. In this study, the focus was on PNP-B, which has not been extensively studied. Bioinformatic analysis was done on the AtPNP-B gene. This included the bioinformatic study of its primary structure, secondary structure, tertiary structure, transcription factor binding sites (TFBS) and its relation to other known proteins. The AtPNP-B gene was shown to be a 510 bp long, including a predicted 138 bp intron. AtPNP-B was also shown to have some sequence similarity with AtPNP-A and CjBAp12. The TFBS for AtPNP-B and OsJPNP-B were compared and they comprised of TFBS that are related to water homeostasis and pathogenesis. This suggested two possible functions; water stress and homeostasis and a pathogenesis related function for PNP-B. Following bioinformatic analysis, the heterologous expression of the AtPNP-B was attempted to investigate whether the AtPNP-B gene encoded a functional protein and to determine the functional role of PNP-B. However, expression was unsuccessful. An evolutionary study was then carried out which revealed that there were some plants without the intron such as, rice, leafy spurge, oilseed rape, onion, poplar, sugar cane, sunflower and tobacco. These plants would therefore be used for expression and functional studies in the future. The evolutionary studies also revealed that PNP-B had a relationship with expansins and the endoglucanase family 45. Other PNP-B related molecules were also obtained from other plant genomes and therefore used in the construction of a phylogenetic tree. The phylogenetic tree revealed that AtPNP-B clustered in the same group as CjBAp12 while AtPNP-A had its own cluster group. There were also other PNP-B like molecules that clustered in the same group as expansins (α- and β-). Thus, we postulate that, like PNP-A, PNP-B also has a possible function in water and salt homeostasis. However, due to the clustering iii of AtPNP-B into the same group as CjBAp12, a possible role of PNP-B in pathogenesis-related response is also postulated.
- Full Text:
- Authors: Hove, Runyararo Memory
- Date: 2009
- Subjects: Plant hormones , Peptides , Plant gene expression , Peptide hormones , Peptides -- Separation
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11251 , http://hdl.handle.net/10353/155 , Plant hormones , Peptides , Plant gene expression , Peptide hormones , Peptides -- Separation
- Description: Plant natriuretic peptides (PNP) are novel peptides which, like in vertebrates, have been shown to have a function associated with water and salt homeostasis. Two PNP-encoding genes have been identified and isolated from Arabidopsis thaliana, namely; AtPNP-A and AtPNP-B. In this study, the focus was on PNP-B, which has not been extensively studied. Bioinformatic analysis was done on the AtPNP-B gene. This included the bioinformatic study of its primary structure, secondary structure, tertiary structure, transcription factor binding sites (TFBS) and its relation to other known proteins. The AtPNP-B gene was shown to be a 510 bp long, including a predicted 138 bp intron. AtPNP-B was also shown to have some sequence similarity with AtPNP-A and CjBAp12. The TFBS for AtPNP-B and OsJPNP-B were compared and they comprised of TFBS that are related to water homeostasis and pathogenesis. This suggested two possible functions; water stress and homeostasis and a pathogenesis related function for PNP-B. Following bioinformatic analysis, the heterologous expression of the AtPNP-B was attempted to investigate whether the AtPNP-B gene encoded a functional protein and to determine the functional role of PNP-B. However, expression was unsuccessful. An evolutionary study was then carried out which revealed that there were some plants without the intron such as, rice, leafy spurge, oilseed rape, onion, poplar, sugar cane, sunflower and tobacco. These plants would therefore be used for expression and functional studies in the future. The evolutionary studies also revealed that PNP-B had a relationship with expansins and the endoglucanase family 45. Other PNP-B related molecules were also obtained from other plant genomes and therefore used in the construction of a phylogenetic tree. The phylogenetic tree revealed that AtPNP-B clustered in the same group as CjBAp12 while AtPNP-A had its own cluster group. There were also other PNP-B like molecules that clustered in the same group as expansins (α- and β-). Thus, we postulate that, like PNP-A, PNP-B also has a possible function in water and salt homeostasis. However, due to the clustering iii of AtPNP-B into the same group as CjBAp12, a possible role of PNP-B in pathogenesis-related response is also postulated.
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