An assessment of the status of psylloid species (Hemiptera: Psylloidea) as potential pests of commercial citrus in southern Africa: implications for pest management
- Authors: Moagi, Raynold
- Date: 2024-10-11
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/464417 , vital:76509
- Description: Psylloids (Hemiptera: Psylloidea), constitute a group of plant sap-sucking insects, some of which are economically significant pests in different ecosystems due to their potential to transmit Gram-negative bacteria, such as the Candidatus Liberibacter species. The African citrus triozid (ACT), Trioza erytreae (Del Guercio), which transmits African citrus greening and the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, which transmits Asian citrus greening are significant threats to citrus. Asian citrus psyllid poses a global economic threat due to its ability to vector “Candidatus Liberibacter asiaticus” (CLas), which can rapidly kill citrus trees. However, both ACP and CLas are currently not present in southern Africa but are present in East and West Africa. In the Afrotropical region, 71 triozid species are known to occur and approximately 41 described Diaphorina species in southern Africa. Currently, two indigenous Diaphorina species, Diaphorina punctulata and Diaphorina zebrana have been documented to feed on citrus. There is a significant knowledge gap regarding the ecological roles of other indigenous psylloid species occurring within the citrus environments. Therefore, this study aimed to: (i) determine the diversity and community structure of psylloid species in citrus environments, and (ii) their host ranges through DNA analysis of gut contents to determine if they fed on citrus. Field surveys were carried out across 12 distinct commercial citrus environments across Limpopo and Mpumalanga provinces between 2022 and 2023. Psylloids were collected using yellow sticky traps and an insect sweep-net. Collected psylloid specimens were preserved in 70% ethanol vials and identified to the lowest possible taxonomic level (i.e. genus or species) using both published and unpublished dichotomous identification keys. Furthermore, citrus leaf samples were collected from the same plants on which psylloids were found in the orchards. Genomic DNA (gDNA) was extracted from both leaf and psylloid samples using two different DNA extraction methods. To confirm if citrus DNA could be detected in the psylloid guts, all leaf gDNA samples were initially amplified using the rbcLaF/R primer pair, targeting a 530-bp region of the chloroplast rbcL gene through the polymerase chain reaction (PCR). Lastly, gut content analysis was performed on 11 psylloid species using the same primer pair through PCR to detect citrus DNA. A total of 4,900 psylloids belonging to five families (i.e. Aphalaridae, Carsidaridae, Liviidae, Psyllidae and Triozidae), 19 genera and 47 species, were collected in citrus environments. More psylloids were recorded in Limpopo (3,754) than in Mpumalanga (1,146). The most abundant species were Pauropsylla trichaeta (1,680), followed by Diaphorina punctulata (466), Trioza erytreae (426), Diaphorina virgata (371), Euryconus sp. (358), Cacopsylla sp. (311), Retroacizzia mopanei (263), Acizzia russellae-group (240), Acizzia sp.3 (216) and Acizzia sp.2 (140). Yellow sticky traps captured 3,265 psylloids in citrus orchards, while an insect sweep-net collected 1,635 psylloids (477 from citrus orchards and 1,158 from adjacent natural vegetation). Data from the insect sweep-net revealed that 22 psylloid species were recorded on citrus. In comparison, nine psylloid species were found on Vachellia spp. and unidentified plant species separately, whereas six, three and two psylloid species were recorded on marula, Ficus sp. and mopane, respectively. The abundance, richness and community structure of psylloids differed significantly between the collection methods, provinces and among plant species. The rbcLaF/R primer pair amplified all citrus leaf gDNA samples, producing amplicons of the targeted 530-bp size. The PCR analysis of 11 psylloid species showed that the rbcLaF/R primer pair amplified plant DNA, with PCR-amplified plant DNA samples producing amplicons between 500-bp and 750-bp in the gut contents of five psyllid species: Diaphorina punctulata, Diaphorina virgata, Diaphorina zebrana, Euryconus sp. and Trioza erytreae. However, the targeted 530-bp plant DNA region was only amplified from the gut contents of Euryconus sp. and Diaphorina punctulata. This study documented psylloid diversity and community structure within commercial citrus environments. The findings indicate that the community of psylloids was diverse in citrus environments, with yellow sticky traps being more effective in monitoring different psyllid species within these environments. Furthermore, the PCR analysis detected citrus DNA in the gut contents of Euryconus sp. and Diaphorina punctulata, suggesting that they could be nibbling on citrus when their specific or main host-plants adjacent to citrus orchards are depleted. However, these insects do not lay their eggs or complete their life cycle on citrus, further confirming that citrus is not their host-plant. Thus, further studies, including Sanger sequencing of PCR-amplified plant DNA, are recommended to confirm the ingested plant species, and host-specific testing including infection trials needs to be conducted. , Thesis (MSc) -- Faculty of Science, Zoology and Entomology, 2024
- Full Text:
- Date Issued: 2024-10-11
- Authors: Moagi, Raynold
- Date: 2024-10-11
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/464417 , vital:76509
- Description: Psylloids (Hemiptera: Psylloidea), constitute a group of plant sap-sucking insects, some of which are economically significant pests in different ecosystems due to their potential to transmit Gram-negative bacteria, such as the Candidatus Liberibacter species. The African citrus triozid (ACT), Trioza erytreae (Del Guercio), which transmits African citrus greening and the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, which transmits Asian citrus greening are significant threats to citrus. Asian citrus psyllid poses a global economic threat due to its ability to vector “Candidatus Liberibacter asiaticus” (CLas), which can rapidly kill citrus trees. However, both ACP and CLas are currently not present in southern Africa but are present in East and West Africa. In the Afrotropical region, 71 triozid species are known to occur and approximately 41 described Diaphorina species in southern Africa. Currently, two indigenous Diaphorina species, Diaphorina punctulata and Diaphorina zebrana have been documented to feed on citrus. There is a significant knowledge gap regarding the ecological roles of other indigenous psylloid species occurring within the citrus environments. Therefore, this study aimed to: (i) determine the diversity and community structure of psylloid species in citrus environments, and (ii) their host ranges through DNA analysis of gut contents to determine if they fed on citrus. Field surveys were carried out across 12 distinct commercial citrus environments across Limpopo and Mpumalanga provinces between 2022 and 2023. Psylloids were collected using yellow sticky traps and an insect sweep-net. Collected psylloid specimens were preserved in 70% ethanol vials and identified to the lowest possible taxonomic level (i.e. genus or species) using both published and unpublished dichotomous identification keys. Furthermore, citrus leaf samples were collected from the same plants on which psylloids were found in the orchards. Genomic DNA (gDNA) was extracted from both leaf and psylloid samples using two different DNA extraction methods. To confirm if citrus DNA could be detected in the psylloid guts, all leaf gDNA samples were initially amplified using the rbcLaF/R primer pair, targeting a 530-bp region of the chloroplast rbcL gene through the polymerase chain reaction (PCR). Lastly, gut content analysis was performed on 11 psylloid species using the same primer pair through PCR to detect citrus DNA. A total of 4,900 psylloids belonging to five families (i.e. Aphalaridae, Carsidaridae, Liviidae, Psyllidae and Triozidae), 19 genera and 47 species, were collected in citrus environments. More psylloids were recorded in Limpopo (3,754) than in Mpumalanga (1,146). The most abundant species were Pauropsylla trichaeta (1,680), followed by Diaphorina punctulata (466), Trioza erytreae (426), Diaphorina virgata (371), Euryconus sp. (358), Cacopsylla sp. (311), Retroacizzia mopanei (263), Acizzia russellae-group (240), Acizzia sp.3 (216) and Acizzia sp.2 (140). Yellow sticky traps captured 3,265 psylloids in citrus orchards, while an insect sweep-net collected 1,635 psylloids (477 from citrus orchards and 1,158 from adjacent natural vegetation). Data from the insect sweep-net revealed that 22 psylloid species were recorded on citrus. In comparison, nine psylloid species were found on Vachellia spp. and unidentified plant species separately, whereas six, three and two psylloid species were recorded on marula, Ficus sp. and mopane, respectively. The abundance, richness and community structure of psylloids differed significantly between the collection methods, provinces and among plant species. The rbcLaF/R primer pair amplified all citrus leaf gDNA samples, producing amplicons of the targeted 530-bp size. The PCR analysis of 11 psylloid species showed that the rbcLaF/R primer pair amplified plant DNA, with PCR-amplified plant DNA samples producing amplicons between 500-bp and 750-bp in the gut contents of five psyllid species: Diaphorina punctulata, Diaphorina virgata, Diaphorina zebrana, Euryconus sp. and Trioza erytreae. However, the targeted 530-bp plant DNA region was only amplified from the gut contents of Euryconus sp. and Diaphorina punctulata. This study documented psylloid diversity and community structure within commercial citrus environments. The findings indicate that the community of psylloids was diverse in citrus environments, with yellow sticky traps being more effective in monitoring different psyllid species within these environments. Furthermore, the PCR analysis detected citrus DNA in the gut contents of Euryconus sp. and Diaphorina punctulata, suggesting that they could be nibbling on citrus when their specific or main host-plants adjacent to citrus orchards are depleted. However, these insects do not lay their eggs or complete their life cycle on citrus, further confirming that citrus is not their host-plant. Thus, further studies, including Sanger sequencing of PCR-amplified plant DNA, are recommended to confirm the ingested plant species, and host-specific testing including infection trials needs to be conducted. , Thesis (MSc) -- Faculty of Science, Zoology and Entomology, 2024
- Full Text:
- Date Issued: 2024-10-11
“COVID-19 made me a single parent”: an interpretative phenomenological analysis of a woman’s perinatal experiences during the COVID-19 pandemic
- Authors: Hadebe, Asanda Locresia
- Date: 2024-04-04
- Subjects: Perinatal care , COVID-19 Pandemic, 2020- Influence , Interpretative phenomenological analysis , Pregnant women South Africa , Pregnancy Psychological aspects , Psychology Qualitative research South Africa
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/435437 , vital:73158
- Description: The COVID-19 pandemic resulted in unprecedented challenges across various aspects of life. In particular, pregnant women encountered unique challenges and circumstances that necessitated adaptation to the experience of the perinatal period1 Satyanarayana et al., (2011). A considerable amount of research has been conducted regarding women's experience of the perinatal period during COVID-19, especially in first world countries. However, inadequate research has been done in the South African context and specifically, there is a lack of qualitative research providing thick descriptions of experience. The analysis describes three master themes supported by subordinate themes. The main themes are (1) A sense of loss and change, (2) Managing COVID-19 and its regulations during the perinatal period, and (3) Glimmers of hope and desirable aftermaths. The study’s findings expand and support the growing literature of women’s experiences on the perinatal period during the COVID-19 pandemic. , Thesis (MA) -- Faculty of Humanities, Psychology, 2024
- Full Text:
- Date Issued: 2024-04-04
- Authors: Hadebe, Asanda Locresia
- Date: 2024-04-04
- Subjects: Perinatal care , COVID-19 Pandemic, 2020- Influence , Interpretative phenomenological analysis , Pregnant women South Africa , Pregnancy Psychological aspects , Psychology Qualitative research South Africa
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/435437 , vital:73158
- Description: The COVID-19 pandemic resulted in unprecedented challenges across various aspects of life. In particular, pregnant women encountered unique challenges and circumstances that necessitated adaptation to the experience of the perinatal period1 Satyanarayana et al., (2011). A considerable amount of research has been conducted regarding women's experience of the perinatal period during COVID-19, especially in first world countries. However, inadequate research has been done in the South African context and specifically, there is a lack of qualitative research providing thick descriptions of experience. The analysis describes three master themes supported by subordinate themes. The main themes are (1) A sense of loss and change, (2) Managing COVID-19 and its regulations during the perinatal period, and (3) Glimmers of hope and desirable aftermaths. The study’s findings expand and support the growing literature of women’s experiences on the perinatal period during the COVID-19 pandemic. , Thesis (MA) -- Faculty of Humanities, Psychology, 2024
- Full Text:
- Date Issued: 2024-04-04
The proprietary consequences of foreign marriages for the purposes of estate planning and succession in South Africa
- Authors: Thorne, Aimee Liza
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/420725 , vital:71773
- Description: This thesis is intended to provide clarity regarding the proprietary consequences of foreign marriages in the context of estate planning and succession in South Africa. It includes an exploration into the impact of specific legislation on foreign marriages, the implications of matrimonial property regimes on a deceased estate in South Africa, private international law principles affecting matrimonial property and succession claims, the scope of antenuptial contracts and postnuptial contracts as existing remedies for foreign remedies, and the harmonisation of succession and matrimonial property law in the European Union (EU). It furthermore suggests potential reforms for the overhaul of the lex domicilii matrimonii principle and associated issues arising therefrom. Various specific issues discussed include the waiver of accrual claims, maintenance claims by surviving spouses of a foreign marriage, choice-of-law rules for matrimonial property rights and succession rights, the registration of a domestic or foreign antenuptial contract for foreign marriages, the formal validity and proper law of antenuptial contracts, the operation of the immutability principle on a domestic and international level, and the common-law approach to the recognition of postnuptial contracts and spousal donations. , Thesis (LLM) -- Faculty of Law, Law, 2023
- Full Text:
- Date Issued: 2023-03-29
- Authors: Thorne, Aimee Liza
- Date: 2023-03-29
- Subjects: Uncatalogued
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/420725 , vital:71773
- Description: This thesis is intended to provide clarity regarding the proprietary consequences of foreign marriages in the context of estate planning and succession in South Africa. It includes an exploration into the impact of specific legislation on foreign marriages, the implications of matrimonial property regimes on a deceased estate in South Africa, private international law principles affecting matrimonial property and succession claims, the scope of antenuptial contracts and postnuptial contracts as existing remedies for foreign remedies, and the harmonisation of succession and matrimonial property law in the European Union (EU). It furthermore suggests potential reforms for the overhaul of the lex domicilii matrimonii principle and associated issues arising therefrom. Various specific issues discussed include the waiver of accrual claims, maintenance claims by surviving spouses of a foreign marriage, choice-of-law rules for matrimonial property rights and succession rights, the registration of a domestic or foreign antenuptial contract for foreign marriages, the formal validity and proper law of antenuptial contracts, the operation of the immutability principle on a domestic and international level, and the common-law approach to the recognition of postnuptial contracts and spousal donations. , Thesis (LLM) -- Faculty of Law, Law, 2023
- Full Text:
- Date Issued: 2023-03-29
Nature conservation, protected areas and local communities : the Tsitsikamma National Park
- Authors: Le Fleur, Yvette Adele
- Date: 2021-04
- Subjects: National parks and reserves -- South Africa -- Eastern Cape , Protected areas -- Management , Programme of Work on Protected Areas , Human ecology -- South Africa -- Tsitsikamma , Nature conservation -- Citizen participation
- Language: English
- Type: thesis , text , Masters , MSocSc
- Identifier: http://hdl.handle.net/10962/177247 , vital:42803
- Description: Since the twentieth century, protected areas, usually in the form of nature reserves or national parks, have become increasingly more dominant as an international conservation strategy. An important factor in protected area management is the relationship between protected areas and its surrounding communities. Historically, the fortress conservation model based on the exclusion of human use and occupation prevailed in relation to protected areas. It is known that this approach brought with it many social and environmental injustices to local communities living in or adjacent to parks. However, a shift in conservation thinking occurred towards the end of the twentieth century, where it has increasingly been advocated for a more participatory approach in protected area management. South Africa has signed the international Convention on Biological Diversity that promotes a participatory approach to nature conservation, which is also reflected in the country’s national laws and policies. In relation to the literature and the lens of political ecology, this thesis sets out to assess in what ways and to what extent the participatory approach has been embraced by South Africa’s conservation authorities. As a case study, it looks at the Tsitsikamma National P ark ( incorporated into the larger Garden Route National Park). In order to get insight on h ow the participatory approach plays itself out at the T NP and what the nature of local communities’ relationship with the park is, this study looks at the aspects of (1) local communities’ socio-economic conditions, (2) their relationship with their natural environment, (3) their perception of nature conservation and (4) their perception of tourism. Then, taking all these aspects into account, (5) how local communities, in general, perceive their relationship with the p ark and its authorities. This study looked at the communities of Kurland, Nature’s Valley, Covie and Storms River. It shows in concurrence with other studies, that despite progressive laws and policies that express the intention of the South African state and SANParks to embrace the participatory approach, its implementation o n t he g round is riddled with challenges. , Thesis (MSocSc) -- Faculty of Humanities, Athropology, 2021
- Full Text:
- Date Issued: 2021-04
- Authors: Le Fleur, Yvette Adele
- Date: 2021-04
- Subjects: National parks and reserves -- South Africa -- Eastern Cape , Protected areas -- Management , Programme of Work on Protected Areas , Human ecology -- South Africa -- Tsitsikamma , Nature conservation -- Citizen participation
- Language: English
- Type: thesis , text , Masters , MSocSc
- Identifier: http://hdl.handle.net/10962/177247 , vital:42803
- Description: Since the twentieth century, protected areas, usually in the form of nature reserves or national parks, have become increasingly more dominant as an international conservation strategy. An important factor in protected area management is the relationship between protected areas and its surrounding communities. Historically, the fortress conservation model based on the exclusion of human use and occupation prevailed in relation to protected areas. It is known that this approach brought with it many social and environmental injustices to local communities living in or adjacent to parks. However, a shift in conservation thinking occurred towards the end of the twentieth century, where it has increasingly been advocated for a more participatory approach in protected area management. South Africa has signed the international Convention on Biological Diversity that promotes a participatory approach to nature conservation, which is also reflected in the country’s national laws and policies. In relation to the literature and the lens of political ecology, this thesis sets out to assess in what ways and to what extent the participatory approach has been embraced by South Africa’s conservation authorities. As a case study, it looks at the Tsitsikamma National P ark ( incorporated into the larger Garden Route National Park). In order to get insight on h ow the participatory approach plays itself out at the T NP and what the nature of local communities’ relationship with the park is, this study looks at the aspects of (1) local communities’ socio-economic conditions, (2) their relationship with their natural environment, (3) their perception of nature conservation and (4) their perception of tourism. Then, taking all these aspects into account, (5) how local communities, in general, perceive their relationship with the p ark and its authorities. This study looked at the communities of Kurland, Nature’s Valley, Covie and Storms River. It shows in concurrence with other studies, that despite progressive laws and policies that express the intention of the South African state and SANParks to embrace the participatory approach, its implementation o n t he g round is riddled with challenges. , Thesis (MSocSc) -- Faculty of Humanities, Athropology, 2021
- Full Text:
- Date Issued: 2021-04
Towards a biological profile for South African perinatal remains: osteological and genetic perspectives
- Authors: Thornton, Roxanne
- Date: 2019
- Subjects: Identification , Forensic osteology , Methylation , RNA , Autopsy
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/68102 , vital:29198 , DOI 10.21504/10962/68102
- Description: Forensic identification of abandoned and suspected infanticide cases admitted to the South African Forensic Pathology Services is often impossible due to decomposition of the remains. In these cases, investigation of suspected criminal activity is almost never pursued. Ancillary tests in the form of anthropological and molecular analyses can assist with the forensic identification of perinatal remains. To provide fundamental information about bone development of perinatal skeleton, osteological and genetic techniques focusing on the pars basilaris, pars lateralis, sternal rib and left femur were used. Samples were obtained from unidentified and unclaimed remains originating from the Johannesburg Forensic Pathology Service (JFPS). To provide a biological age to individuals in the collection, dental aging was used to categorize remains for comparisons with anthropological and molecular data. A molecular protocol was designed to sex individuals using the X-linked G6PD and Y-linked SRY genes. Bone development was studied using osteometric and morphological data of dry bone remains coupled with bone mineral density analysis (Micro-CT). The methylation levels of CpG rich sites within the promoter region of selected bone-associated genes were incorporated to examine silencing of genes during development. Osteological results support the use of the pars basilaris, pars lateralis and femur for age-at-death estimations as well as provide the foundation for dry bone aging criteria for South African individuals. Data compared with established skeletal aging standards indicated developmental differences between populations. Through the use of animal models and the perinatal sternal rib tissue, insights and precautions into the use of post mortem bone derived RNA for forensic applications is communicated. The methylation status of CpG rich sites within the promoter regions support the hypothesis for interdependent machinery involving selected genes during early bone development. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2019
- Full Text:
- Date Issued: 2019
- Authors: Thornton, Roxanne
- Date: 2019
- Subjects: Identification , Forensic osteology , Methylation , RNA , Autopsy
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/68102 , vital:29198 , DOI 10.21504/10962/68102
- Description: Forensic identification of abandoned and suspected infanticide cases admitted to the South African Forensic Pathology Services is often impossible due to decomposition of the remains. In these cases, investigation of suspected criminal activity is almost never pursued. Ancillary tests in the form of anthropological and molecular analyses can assist with the forensic identification of perinatal remains. To provide fundamental information about bone development of perinatal skeleton, osteological and genetic techniques focusing on the pars basilaris, pars lateralis, sternal rib and left femur were used. Samples were obtained from unidentified and unclaimed remains originating from the Johannesburg Forensic Pathology Service (JFPS). To provide a biological age to individuals in the collection, dental aging was used to categorize remains for comparisons with anthropological and molecular data. A molecular protocol was designed to sex individuals using the X-linked G6PD and Y-linked SRY genes. Bone development was studied using osteometric and morphological data of dry bone remains coupled with bone mineral density analysis (Micro-CT). The methylation levels of CpG rich sites within the promoter region of selected bone-associated genes were incorporated to examine silencing of genes during development. Osteological results support the use of the pars basilaris, pars lateralis and femur for age-at-death estimations as well as provide the foundation for dry bone aging criteria for South African individuals. Data compared with established skeletal aging standards indicated developmental differences between populations. Through the use of animal models and the perinatal sternal rib tissue, insights and precautions into the use of post mortem bone derived RNA for forensic applications is communicated. The methylation status of CpG rich sites within the promoter regions support the hypothesis for interdependent machinery involving selected genes during early bone development. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2019
- Full Text:
- Date Issued: 2019
Bioprospecting for amylases, cellulases and xylanases from ericoid associated fungi, their production and characterisation for the bio-economy
- Authors: Adeoyo, Olusegun Richard
- Date: 2018
- Subjects: Mycorrhizal fungi , Hydrolases , Ericaceae South Africa , Ericaceae Molecular aspects
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/64327 , vital:28533
- Description: South Africa is one of the most productive areas for ericaceous plants with about 850 identified species in the Cape Floral Region. The Albany Centre of Endemism where all fungi used in this study were isolated from, falls within this region. Ericaceous plants interact with some fungi via an association called the ericoid mycorrhizal (ERM) association. All fungi used in this study were isolated from roots of six ericaceous plants; Erica cerinthoides, Erica demissa, Erica chamissonis, Erica glumiflora, Erica caffra and Erica nemorosa. Fungal enzymes are known to play a significant role in the food, brewing, detergent, pharmaceutical and biofuel industries. The enzyme industry is among the major sectors of the world, and additional novel sources are being explored from time to time. This study focussed on amylases (amyloglucosidase, AMG), cellulases (endoglucanase) and xylanases (endo-1,4-P-xylanase) production from ERM fungal isolates. Out of the fifty-one (51), fungal isolates screened, ChemRU330 (Leohumicola sp.), EdRU083 and EdRU002 were among the fungi that had the highest activities of all the enzymes. They were tested for the ability to produce amylases and cellulases under different pH and nutritional conditions that included: carbon sources, nitrogen sources and metal ions, at an optimum temperature of 28°C in a modified Melin-Norkrans (MMN) liquid medium. Cellulase specific activity of 3.99, 2.18 and 4.31 (U/mg protein) for isolates EdRU083, EdRU002 and ChemRU330, respectively, was produced at an optimal pH of 5.0. For amylase, ChemRU330 had the highest specific activity of 1.11 U/mg protein while EdRU083 and EdRU02 had a specific activity of 0.80 and 0.92 U/mg protein, respectively, at the same pH with corresponding biomass yield of 113, 125 and 97 mg/50 ml, respectively. Increased enzyme activities and improved mycelial biomass production were obtained in the presence of supplements such as potassium, sodium, glucose, maltose, cellobiose, tryptone and peptone, while NaFe-EDTA and cobalt inhibited enzyme activity. ChemRU330 was selected to determine the consistency and amount of amylase, cellulase and xylanase formed after several in vitro subculturing events. AMG and endo-1,4-P-xylanase were found to have the most consistent production throughout the study period. The AMG was stable at 45oC (pH 5.0), retaining approximately 65% activity over a period of 24 h. The molecular mass of AMG and endo-1,4-P-xylanase were estimated to be 101 kDa and 72 kDa, respectively. The Km and kcat were 0.38 mg/ml and 70 s-1, respectively, using soluble starch (AMG). For endo-1,4-P-xylanase, the Km and Vmax were 0.93 mg/ml and 8.54 U/ml, respectively, using beechwood xylan (endo-1,4-P-xylanase) as substrate. Additionally, crude extracts of five root endophytes with unique morphological characteristics were screened for antibacterial properties and was followed by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). L. incrustata (ChemRU330) and Chaetomium sp. extracts exhibited varying degrees of inhibition against two Gram-positive and Gram-negative bacteria. The crude extract of L. incrustata was the most effective which was found to inhibit Staphylococcus aureus (MIC: 1 mg/ml), Bacillus subtilis (MIC: 2 mg/ml) and Proteus vulgaris (MIC: 16 mg/ml). The L. incrustata displayed potential for antibacterial production and could be considered as an additional source of new antimicrobial agents in drug and food preservation. Also, the three isolates used for enzyme production were identified to genus and species levels, i.e., Leohumicola incrustata (ChemRU330), Leohumicola sp. (EdRU083) and Oidiodendron sp. (EdRU002) using both ITS and Cox1 DNA regions. The molecular analysis results indicated that these ERM mycorrhizal fungi were similar to those successfully described by some researchers in South Africa and Australia. Therefore, this study opens new opportunities for exploring ERM fungal biomolecules for the bio-economy. The promising physicochemical properties, starch and xylan hydrolysis end- products, and being non-pathogenic make AMG and endo-1,4-P-xylanase potential candidates for future applications as additives in the food industry for the production of glucose, glucose syrups, high-fructose corn syrups, and as well as the production of bioethanol. Finally, the findings of this study revealed that it is possible to produce hydrolytic enzymes from ERM fungi in vitro using chemically defined media. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
- Authors: Adeoyo, Olusegun Richard
- Date: 2018
- Subjects: Mycorrhizal fungi , Hydrolases , Ericaceae South Africa , Ericaceae Molecular aspects
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/64327 , vital:28533
- Description: South Africa is one of the most productive areas for ericaceous plants with about 850 identified species in the Cape Floral Region. The Albany Centre of Endemism where all fungi used in this study were isolated from, falls within this region. Ericaceous plants interact with some fungi via an association called the ericoid mycorrhizal (ERM) association. All fungi used in this study were isolated from roots of six ericaceous plants; Erica cerinthoides, Erica demissa, Erica chamissonis, Erica glumiflora, Erica caffra and Erica nemorosa. Fungal enzymes are known to play a significant role in the food, brewing, detergent, pharmaceutical and biofuel industries. The enzyme industry is among the major sectors of the world, and additional novel sources are being explored from time to time. This study focussed on amylases (amyloglucosidase, AMG), cellulases (endoglucanase) and xylanases (endo-1,4-P-xylanase) production from ERM fungal isolates. Out of the fifty-one (51), fungal isolates screened, ChemRU330 (Leohumicola sp.), EdRU083 and EdRU002 were among the fungi that had the highest activities of all the enzymes. They were tested for the ability to produce amylases and cellulases under different pH and nutritional conditions that included: carbon sources, nitrogen sources and metal ions, at an optimum temperature of 28°C in a modified Melin-Norkrans (MMN) liquid medium. Cellulase specific activity of 3.99, 2.18 and 4.31 (U/mg protein) for isolates EdRU083, EdRU002 and ChemRU330, respectively, was produced at an optimal pH of 5.0. For amylase, ChemRU330 had the highest specific activity of 1.11 U/mg protein while EdRU083 and EdRU02 had a specific activity of 0.80 and 0.92 U/mg protein, respectively, at the same pH with corresponding biomass yield of 113, 125 and 97 mg/50 ml, respectively. Increased enzyme activities and improved mycelial biomass production were obtained in the presence of supplements such as potassium, sodium, glucose, maltose, cellobiose, tryptone and peptone, while NaFe-EDTA and cobalt inhibited enzyme activity. ChemRU330 was selected to determine the consistency and amount of amylase, cellulase and xylanase formed after several in vitro subculturing events. AMG and endo-1,4-P-xylanase were found to have the most consistent production throughout the study period. The AMG was stable at 45oC (pH 5.0), retaining approximately 65% activity over a period of 24 h. The molecular mass of AMG and endo-1,4-P-xylanase were estimated to be 101 kDa and 72 kDa, respectively. The Km and kcat were 0.38 mg/ml and 70 s-1, respectively, using soluble starch (AMG). For endo-1,4-P-xylanase, the Km and Vmax were 0.93 mg/ml and 8.54 U/ml, respectively, using beechwood xylan (endo-1,4-P-xylanase) as substrate. Additionally, crude extracts of five root endophytes with unique morphological characteristics were screened for antibacterial properties and was followed by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). L. incrustata (ChemRU330) and Chaetomium sp. extracts exhibited varying degrees of inhibition against two Gram-positive and Gram-negative bacteria. The crude extract of L. incrustata was the most effective which was found to inhibit Staphylococcus aureus (MIC: 1 mg/ml), Bacillus subtilis (MIC: 2 mg/ml) and Proteus vulgaris (MIC: 16 mg/ml). The L. incrustata displayed potential for antibacterial production and could be considered as an additional source of new antimicrobial agents in drug and food preservation. Also, the three isolates used for enzyme production were identified to genus and species levels, i.e., Leohumicola incrustata (ChemRU330), Leohumicola sp. (EdRU083) and Oidiodendron sp. (EdRU002) using both ITS and Cox1 DNA regions. The molecular analysis results indicated that these ERM mycorrhizal fungi were similar to those successfully described by some researchers in South Africa and Australia. Therefore, this study opens new opportunities for exploring ERM fungal biomolecules for the bio-economy. The promising physicochemical properties, starch and xylan hydrolysis end- products, and being non-pathogenic make AMG and endo-1,4-P-xylanase potential candidates for future applications as additives in the food industry for the production of glucose, glucose syrups, high-fructose corn syrups, and as well as the production of bioethanol. Finally, the findings of this study revealed that it is possible to produce hydrolytic enzymes from ERM fungi in vitro using chemically defined media. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
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