A research report submitted to the Faculty of Science, University of the Witwatersrand, in partial fulfilment of the requirements for the degree of Master of Science Johannesburg, August 2015 / The central Kalahari is likely to become hotter and drier as a result of climate change in the region. These changes may result in behavioural changes in Gemsbok due to temperature induced stress, in spite of physiological and behavioural adaptations, and are likely to manifest in a preference for particular landscape patch classes. Recent Landsat 8 satellite imagery and classification analysis were used to map landscape patch classes in a heterogeneous landscape in the central Kalahari. The classification map of the research area identified 6 classes of landscape patches used by Gemsbok. Eight collared Gemsbok were tracked by satellite to monitor their movement in their respective home ranges over a period of 9 months. Gemsbok locations were plotted on to the classification map, and location frequency distributions were produced for each landscape patch class. Gemsbok home ranges were calculated using minimum convex polygon geometry, and the available patch class areas within each home range were analysed against the usage patterns of each animal. The analysis showed less preference for shade producing classes and more preference for open classes. Exploration of the role of temperature in landscape patch selection showed that temperature is a weak predictor of patch class, that critical temperature thresholds have not yet been reached, and that Gemsbok preference for pans is more likely related to seasonally available forage and reduced predation risk in a “landscape of fear” (Laundré, Hernández, & Altendorf, 2001).

Terfezia pfeilii is an edible mycorrhizal fungus that thrives in the Kalahari Desert of southern Africa. It is best known by desert dwellers for its flavour and as a source of nutrition. Although the genus Terfezia is generally regarded as being an ectomycorrhizal mycobiont, the exact mycorrhizal type formed by T. pfeilli and its' associated host plants remains uncertain. Discovery of the host plants for T. pfeilii would first be required in order to further investigate the life cycle and cultivation of this truffle. This study focussed on the isolation of mycelia from the ascocarp, optimising the growth conditions of the mycelial cultures, rapid molecular identification of T. pfeilii, investigation of potential helper bacteria and mycorrhizal synthesis experiments. T. pfeilii ascocarps were harvested from the Spitskop Nature Reserve in Upington, South Africa. Ascocarps were successfully identified using both morphological and molecular methods. Despite the delayed growth mostly caused by contaminating microorganisms, the isolation of T. pfeilii mycelia culture was successful. Molecular techniques were used to confirm the identity of the pure culture. Further studies were conducted on ways to improve the growth conditions of the mycelial culture on Fontana medium. An optimum temperature of 32°C, the addition of Bovine Serum Albumin as a nitrogen source and a pH of 7.5 significantly improved the growth of T. pfeilii in vitro. A rapid PeR-based molecular method was developed to speed up the identification of T. pfeilii. Specific primers that can exclusively amplify the ITS region of T. pfeilii were designed and used to identify both the ascocarps and the mycelial culture. The specificity of these primers was confirmed by their inability to amplify DNA from the isolates of contamining fungi obtained during the isolation process. Molecular comparison was made to confirm the reclassification of South African samples of T. pfeilii as Kalaharituber pfeilii as proposed by Ferdman et al.,(2005). However, in this study, the name T. pfeilii has been retained. A total of 17 bacterial isolates were obtained from the fruiting bodies of T. pfeaii and these were tested for stimulation of mycelial growth in vitro, indole production and phosphate solubilising capabilities. Bacterial isolates that showed potential to be Mycorrhization Helper Bacteria (MHB) were identified as Paenibacillus sp., Bacillus sp. and Rhizobium tropici. Selected plant seedlings were inoculated with T. pfeilii cultures or ascocarp slurry in order to re-establish the mycorrhizal association. After 8 months, light microscopy observations revealed an endomycorrhizal type association between Cynodon dactylon and T. pfeilii. This was confirmed with molecular analysis using specific T. pfeilii ITS primers. After 15 months, molecular methods confirmed Acacia erioloba as another host plant. These results have provided essential information paving the way for further investigation into the life cycle and biology of the Kalahari truffle. / KMBT_363 / Adobe Acrobat 9.53 Paper Capture Plug-in