Orchid mycorrhizal and endophytic fungal diversity of three co-occurring terrestrial orchids in the large African genus Disa (Orchidaceae)
- Authors: Khambule, Nondumiso Venessia
- Date: 2020
- Subjects: Orchids South Africa , Mycorrhizal fungi South Africa , Endomycorrhizas South Africa , Endophytes , Orchids Roots
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/163341 , vital:41030
- Description: Orchids (in the family Orchidaceous) are one of the richest plant families and approximately 500 species are found in South Africa. A number of orchid species are found on disturbed areas and many of the terrestrial species grow in poor soils with low mineral nutrient availability. Most orchid species are thought to be associated with mycorrhizal fungi for germination and mycorrhiza provides nutrients for the survival of adult plants. The aim of this study was to select Orchidaceous plant species and to isolate, identify and characterize the orchid endophytes and assess these isolates for potential antimicrobial and enzymatic activities Isa is the largest genus in South Africa and three Disa species co-occurring in a small geographical area were selected. These included Disa bracteata, D. cornuta and D. polygonoides which span three sections of the genus. Roots were stained to confirm the mycorrhizal status of the Disa species. Mycorrhizal pelotons structures were microscopically observed inside root cells. The presence of pelotons is indictive of mycorrhizal fungal interactions within the orchid roots and areas associated with the site of nutrient exchange between plant and fungus. The presence of pelotons, however, does not give n indication of the fungal species involved. The endophytes were successfully isolated in pure cultures on potato dextrose agar (PDA). All slow growing isolates were selected, and further molecular identification undertaken; DNA was extracted, and PCR amplified using internal transcribed spacer (ITS1F and ITS4) fungal primers. The amplified products were then sequenced and analysed by comparison to sequences in the GenBank database. Trichoderma, Penicillium, Metapochonia, Talaromyces, Oidiodendron Neopestalotiopsis, and Chaetomium were identified from these sequences. The presence of other fungal root endophytes was suspected despite the rigorous surface sterilization procedure used. The primers used to amplify the ITS region are the universal barcoding primers which are specific to fungi. ITS1F is one of the primers designed to amplify a broad range of fungi. DNA was extracted from orchid roots and amplicons were cloned into a pGEMT plasmid vector. Individual clones were sequenced and aligned with Mega software and compared to sequences in the GenBank and UNITE database. Based on percentage sequence identity, unidentified Tulasnella species, Tullasnela colaspora, and various Ascomycota endophytes were identified as contributing to the endophytic root fungal diversity of the selected Disa species. The Disa species investigated in this study were associated with several soil endophytes. D. bracteata, D. polygonoides were collected from the same site along the road verge which is regarded as being disturbed. Based on both culture – dependent and independent techniques employed Oidiodendron was found associated with both species. Antimicrobial activity was determined using a well diffusion method using extracts from the isolated fungi against the bacterial isolates Bacillus cereus, Staphylococcus aureus, Escherichia coli and Pseudomonas puptida. Most of the isolated fungi showed at least one potential inhibition effect against one of the bacterial isolates. The extracts that showed potential antimicrobial activity could be further screened to determine the compounds produced as secondary metabolites using techniques such as LC-MS Enzymatic activities of protease, cellulose and amylase were determined using solid media amended with milk protein, carboxymethylcellulose (CMC) and starch. The majority of fungal isolates tested positive with amylase and cellulose with only a few fungal isolates testing positive for protease activity. Broth cultures containing CMC and starch were shown to enhance biomass production in approximately 40 % of the fungal isolates. Degradation of the substrates is required in order to provide carbon to the fungus under test in order to optimize fungal growth as well as to gain insight into their ecological role. Enzyme activity was evident particularly when cellulose and starch were provided as substrates. All the fungal isolates tested grew on the amended medium, with 40% of the isolates preferring to utilize CMC and/or starch, indicating the ability of these fungi to utilize various resources for carbon acquisitions. , Thesis (PhD) -- Faculty of Science, Biochemistry and Microbiology, 2020
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Reproductive biology of important invasive plants in the Eastern Cape Province of South Africa
- Authors: Thorne, Cara-Jayne
- Date: 2016
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/3683 , vital:20535
- Description: Baker’s rule predicts species capable of uniparental reproduction are more likely to establish after long distance dispersal (or introduction in the case of invasive plants), thus the ability to undergo autonomous self-pollination should promote colonization. An investigation into the reproductive biology of eleven invasive species in the Eastern Cape Province of South Africa between 2012 and 2014 showed strong support for Baker’s law. Breeding system results showed evidence of self-compatibility in eight species, ten species were capable of autonomous self-pollination, and only three species showed evidence of varying levels of self-incompatibility. These results provide evdence that autonomous self-pollination occurs more frequently among invasive species, self-compatible species more frequently become invasive compared with self-incompatible species and that autogamous species should have a larger invasive range. Co-opting suitable local pollinators may be problematic for invasive plants, especially those with highly specialized pollination systems. The species investigated appear to be largely independent of pollinators, mostly setting seed in the absence of pollinators. Despite this, all eleven invasive plant species are regularly visited by a variety of generalist pollinators including Apis mellifera, Xylocopa bees and Allodapini species. Pollinators all carried substantial pollen loads, even managing to extract pollen from more specialized plants, such as the poricidal anthers of Solanum species, and frequently came into contact with both anthers and stigmas. Considering most of theses species are capable of autonomous self-pollination, their reliance on pollinators may be low. Even the most self-incompatible species, Passiflora caerulea, appears to have successfully co-opted local pollinators (honeybees and carpenter bees), ensuring successful pollination and seed set. The ability for cross-pollination by local pollinators allows for some degree of genetic variation within invasive plant populations, especially for self-incompatible species. Inadequate pollen deposition by unreliable or inefficient pollinators, or reduced resource availability, can result in pollen limitation. Invasive plant species may be especially susceptible, with three of the plant species investiagted showing possible pollen limitation at the level of seed set, while the remaining nine invasive plant species showed no evidence of pollen limitation in South Africa. The ability to utilize uniparental reproduction (as Baker’s law predicted), and co-opt local pollinators has allowed invasive plants in the Eastern Cape to successfully establish and persist after introduction. Hence, the inclusion of reproductive traits of plants should therefore be included in risk assessments for future plant invaders.
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Variation in breeding systems, floral morphology and nectar properties in three co-occurring Erica species with contrasting pollination syndromes
- Authors: Lombardi, Giorgio Colombo
- Date: 2015
- Subjects: Ericas -- South Africa -- Western Cape , Ericas -- Morphology , Ericas -- Reproduction , Ericas -- Pollination , Pollination by animals , Pollination by insects , Nectar
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4273 , http://hdl.handle.net/10962/d1018567
- Description: The genus Erica is the most species rich in the Cape Floristic Kingdom, yet there are limited data on the various flower-pollinator interactions and breeding systems for the majority of these species. Until recently research has focussed on predictive floral traits, including flower shape, length of corolla and corolla opening to identify likely pollinators in this genus. Field observations provide an empirical test of such predictions. This study investigated three species of Erica and compared their pollination and breeding systems and floral biology. Research, including field experiments and pollinator observations was undertaken in the Vogelgat Private Nature Reserve, Maanschynkop Nature Reserve and Boskloof farm in the vicinity of Hermanus in the Western Cape Province of South Africa. Field observations were conducted to determine what flower- visitor interactions occurred, while nectar volumes and sugar concentrations were measured to determine the value of the reward to the different visitors. Selective exclusion and breeding system experiments were carried out to determine whether these Erica species were capable of autonomous self-fertilization or whether they are pollinator dependent for breeding success. The unusual morphology of Erica lanuginosa suggested rodent-pollination. Field observations, including photographs of visits to plants in the field, the presence of pollen in scat sample and selective exclusion and breeding system experiments identified rodents, primarily Acomys subspinosus to be the primary pollinator. Long-proboscid flies of the family Nemestrinidae were found, on the basis of field observations to be responsible for pollination of the endemic Erica aristata. This was supported by with nectar volume and sugar concentration samples which are consistent with other long-proboscid fly-pollinated plant species. Selective exclusion and breeding system experiments undertaken confirmed that Erica aristata required a pollinator to set seed. Observations and breeding trials revealed bird-pollination in Erica sessiliflora. Nectar volume and sugar concentrations in Erica sessiliflora were in line with other sunbird-pollinated plant species, providing the necessary rewards for sunbirds visiting this species. Specialised pollination by single pollinators was found in all three species and results from breeding system experiments show that out-crossing is important. Further research into pollinator-flower interactions in the genus Erica is necessary, not least to understand more fully the conservation importance of specific pollinators.
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Pollinator mediated selection in Pelargonium reniforme Curtis (Geraniaceae) : patterns and process
- Authors: De Wet, Leigh-Ann Robynne
- Date: 2013-06-25
- Subjects: Pelargoniums -- Classification , Pelargoniums -- Pollen , Pelargoniums -- Selection
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4255 , http://hdl.handle.net/10962/d1008185 , Pelargoniums -- Classification , Pelargoniums -- Pollen , Pelargoniums -- Selection
- Description: Pelargonium reniforme is currently divided into two subspecies, P. reniforme subsp. reniforme and P. reniforme subsp. velutinum. The species falls into section Reniformia along with the closely-related P. sidoides. Observations of the section showed some discrepancies in the current taxonomy; mainly floral variation that was not recorded in the descriptions of the subspecies of P. reniforme, particularly the differences in hypanthium lengths. Patterns of variability were analysed using both morphometrics and inter simple sequence repeat (ISSR) data for P. reniforme (both subspecies), and P. sidoides. Results showed no support for the current subspecific division of P. reniforme but also no support for the current status of P. sidoides as a separate species. However, both morphometric and ISSR data show some evidence for the existence of two distinctly separate groups within P. reniforl1le subsp. velutinum as two distinct types have been recognized: those with bright pink flowers with long tubes (up to 70mm), and those with pale pink flowers with shorter tubes (as little as 8mm). These two forms have been found in a single population (Grahamstown), where they occur sympatrically, as well as in monomorphic populations. Analyses of the processes thought to be responsible for the observed patterns were conducted on the Grahamstown population. Observations of pollinators suggest that long-tongued insects (Butterflies, Nemestrinid flies) pollinate the long-tubed flowers and short-tongued insects (Bombyliid flies, long-tongued bees) pollinate short-tubed flowers. However, analysis of selection gradients in the population, indicate no directional selection is occurring for hypanthium lengths. The results of this thesis show that selection is occurring within Pelargonium reniforme, but at this time, selection is not strongly directional and floral differences are maintained, even when floral forms occur sympatrically. It is suggested that a review of the taxonomy be undertaken as well as additional pollination and selection studies to confirm suspected taxonomic groupings and relationships between the floral forms respectively. , KMBT_363 , Adobe Acrobat 9.54 Paper Capture Plug-in
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