Thesis (MTech (Horticulture)--Cape Peninsula University of Technology, 2017. / A study was conducted to investigate the effects of various media composition and wounding treating on the in vitro propagation of Argyroderma subalbum and A. testiculare explants derived from mature plants, antioxidants and plant growth regulators (PGR) concentrations. One experiment consisted of 3 medium types including Murashige and Skoog (MS) medium strength, vitamin supplement. Fifteen replicates were used for each treatment. The shoots were then sub-cultured to ten replicate regenerated medium consisting of varying levels and combination of indole-3-acetic acid (IAA) and 10 μM 6-Benzyladenine (BA) supplements. In another experiment consisted of varying levels of auxins with MS medium strength, activated charcoal (AC) and vitamin supplements ten replicates were used for each treatment. Results indicated the positive role of cytokinins types’ 6-Benzyladenine (BA), 2-isopentyladenine (2iP) and Kinetin in inducing callus formation from wounded explants. The highest rate of friable callus formation of wounded explants was observed in media containing vitamin supplementation with BA at 10 μM. Callus formation significantly increased with the addition of vitamins at 10 μM on BA, 2iP and kinetin. With regards to the effects of various media composition and wounding explants on in vitro growth and regeneration of A. subalbum and A. testiculare, significant results were achieved with BA, 2iP and kinetin concentrations on explants discoloration and callus formation. The antioxidant treatment, AC did not reduce explants discoloration, but the induction of the callus was developed furthermore, results showed that IAA with BA concentrations without addition of AC there was significantly difference on both species but A. subalbum dominated with browning intensity (Chapter 3). Only sub-culturing of the explants succeeded in preventing explants discoloration and subsequently increased the number of shoots. The interaction between Indole-3-acetic acid (IAA) concentrations combined with BA resulted in the most effective technique in reducing explants discoloration at the media contact point. This study provides an insight into the contributing factor and methods of overcoming the major problem of phenolic oxidation and promoting the in vitro growth and regeneration of A. subalbum and A. testiculare.

Thesis (MTech (Horticulture))--Cape Peninsula University of Technology, 2018. / Drosanthemum micans and Drosanthemum hallii are endangered succulent shrubs of horticultural and medicinal value. They are restricted to the Succulent Karroo, which is one of the world’s biodiversity hotspots. The species risk extinction from illegal over-harvesting for water-wise gardens, erosion by occasional flush floods from ephemeral rivers, competition from alien invasive species, overgrazing and clearing of land for agriculture and human settlement. Although seeds and cuttings may be used in propagating these species, they often require seasonal collection and planting and cuttings struggle to establish, hence the need for in-vitro propagation as an alternative solution. Thus, the main objective of the study was to develop a method for rapid in-vitro shoot and root multiplication and acclimatization of D. micans and D. hallii. To initiate shoot formation, disinfected leaf and stem nodal explants were cultured in Murashige and Skoog (1962) media supplemented with different rates (0, 10, 20 or 30μM) of 2-isopentyladenine, 6-Benzyladenine and kinetin for D. hallii or 2-isopentyladenine, 6-Benzyladenine and Thiadiazuron for D. micans. Shoots from explants were rooted in varying rates (0, 10, 20 or 30μM) of IAA for root initiation. Three media, which were used in previous studies, were tested for acclimatization of rooted explants in i) vermiculite, ii) sand (50%): vermiculite (50%) or iii) sand (75%): perlite (25%). For quantitative evaluation of plant stress, chlorophyll fluorescence index (Fv/Fm) was measured as a proxy for plant stressf stress. It emerged that stem nodal explants of D. hallii tend to produce multiple shoots whilst leaf explants tended to produce callus when cultured in full-strength Murashige and Skoog (1962). Shoot multiplication was optimal in both D. hallii and D. micans at 10 μM of kinetin. Root formation in both D. hallii and D. micans only occurred when shoots were transferred to a full-strength Murashige and Skoog (1962) media without any phytohormones added. The intensity of tissue browning increased at higher levels of cytokinins, suggesting an interaction of plant growth regulators with exudates from explants. Different acclimatization media tested showed no significant differences in the level of stress (Fv/Fm). It is recommended that Murashige and Skoog (1962) media with10 μM kinetin be used for shoot development and multiplication, followed by transfer of the shoots to fresh full-strength Murashige and Skoog (1962) media without hormones for root development. Acclimatization of the rooted explants was possible in one of the following media: i) vermiculite, ii) sand (50%): vermiculite (50%) or iii) sand (75%): perlite (25%) and in a misted greenhouse (ca. 60% RH), with gradual weekly reductions in humidity by 10% over 2 weeks.