Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The generally low concentrations of P and N in the soil, causes most plants to experience nutrient deficiency during their life cycle. Lupins can rely on both cluster roots and nodules for P acquisition and biological nitrogen fixation (BNF) respectively. The legume Lupinus albus is able to survive under low nutrient conditions, because it has two specialized belowground organs for the acquisition of N and P. In this regard, cluster roots increase P uptake and root nodules acquire atmospheric N2 via biological nitrogen fixation (BNF). Although these organs normally tolerates low P conditions, very little is known about their physiological and metabolic flexibility during variations in P supply. Furthermore, the resource allocation (C, N and P) between cluster roots and nodules has also been largely understudied. The aim of this investigation was therefore to determine the resource allocation, physiological and metabolic flexibility of these organs during variations in P supply. Although variation on P supply had no effect on the total biomass, there were significant differences in specialised below-ground organ allocation to cluster roots and nodule formation. Cluster root formation and the associated C-costs increased during low P supply. In contrast to the cluster root decline at high P supply, there was an increase in nodule growth allocation and corresponding C-costs. Since cluster roots were able to increase P acquisition under low P conditions, this below-ground investment may also have benefited the P nutrition of nodules. These findings provide evidence that when lupins acquire N via BNF in their nodules, there may be a trade-off in resource allocation between cluster roots and nodules. The short-term elevated P supply, caused an increased allocation of C and respiratory costs to nodules, at the expense of cluster roots. This alteration was also reflected in the increase in nodule enzyme activities related to organic acid synthesis, such as Phosphoenol-pyruvate Carboxylase (PEPC), Pyruvate Kinase (PK), Malate Dehydrogenase (NADH-MDH) and Malic Enzyme (ME). In cluster roots, the elevated P conditions, caused a decline in these organic acid synthesizing enzymes. This suggests that during short-term elevated P supply, there is a great degree of physiological and metabolic flexibility in the lupin nutrient acquiring structures. / AFRIKAANSE OPSOMMING: Die algemeen lae konsentrasies van fosfaat en stikstof in die grond , veroorsaak dat die meeste plante voedingstekorte ervaar tydens hul lewensiklus. Lupiene kan staatmaak op beide groep-wortels en wortel-knoppies vir P verkryging en biologiese stikstofbinding onderskeidelik. Die peulplant Lupinus albus is in staat om te oorleef onder lae voedings toestande , as gevolg van die twee gespesialiseerde ondergrondse organe vir die verkryging van stikstof en fosfaat. In hierdie verband verhoog groep-wortels fosfaat opname en wortel-knoppies verkry atmosferiese stikstof via biologiese stikstofbinding. Alhoewel hierdie organe normaalweg lae fosfaat toestande verdra , is baie min bekend oor hul fisiologiese en metaboliese buigsaamheid tydens variasies in fosfaat aanwending. Daar is verder ‘n tekort aan die studie van hulpbron toekenning tussen groep-wortels en wortel-knoppies. Die doel van hierdie ondersoek was dus om die toekenning van hulpbronne , fisiologiese en metaboliese buigsaamheid van hierdie organe tydens variasies in fosfaat aanwending te bepaal. Variesie in fosfaat verskaffing het geen invloed op die totale plant biomassa gehad nie, maar daar was wel ‘n beduidende verskil in gespesialiseerde ondergrondse toekenning tussen groep- wortels en wortel-knoppies. Groep-wortel vorming en die gepaardgaande koolstof koste het toegeneem met lae fosfaat verskaffing. In teenstelling met die groep-wortel daling met hoë fosfaat verskaffing, was daar 'n toename in groei van wortel-knoppies en die ooreenstemmende koolstof koste daarvan. Aangesien groep-wortels in staat was om fosfaat verkryging te verhoog onder lae fosfaat toestande, mag hierdie ondergrondse belegging bygedra het tot die voeding van wortel-knoppies . Hierdie bevindings bewys dat lupiene afhanklik van wortel-knoppies ‘n wisselwerking in toekenning van hulpbronne, tussen groep-wrotels en wortel-knoppies handaaf. Kort termyn verhoogde fosfaat aanwending veroorsaak 'n verhoogde toekenning van koolstof en respiratoriese energie na wortel-knoppiess, ten koste van groep-wortels . Hierdie verandering is ook weerspieël in die toename in wortel-knoppie ensiem aktiwiteit in verband met organiese suur sintese (PEPC PK,MDH,ME) . In groep-wortels, het die verhoogde P toestande verder 'n afname in die organiese suur produserende ensieme veroorsaak. Dit dui aan dat tydens kort termyn verhoogde P aanwending, daar 'n groot mate van fisiologiese en metaboliese buigsaamheid in die lupiene voedingstowwe verkryging strukture plaasvind.

Thesis (PhD (Botany and Zoology))—University of Stellenbosch, 2009. / The genus Oxalis forms a major part of the flora of southern Africa, in particular the Cape Floristic Region (CFR) at the southwestern tip of the continent, but the current taxonomy is outdated and ecological knowledge of the lineage is sadly incomplete. In this thesis I set out to address several aspects of Oxalis systematics that urgently require attention. Firstly, the current macro-morphological taxonomy requires phylogenetic testing, as it is acknowledged to be incomplete and artificial. I address this need by providing a DNA sequence-based phylogeny of three markers, using three different inference methods, for nearly three quarters of the indigenous species. This phylogeny confirmed both the monophyly of the southern African taxa, and the artificiality of the current classification system. It is congruent with previous sequence-based reconstructions of smaller groups of southern African Oxalis species, and with the palynological classification proposed for the genus. Secondly, previous phylogenetic work on the southern African members could not resolve basal relationships within the southern African clade. I attempt to address this problem by sequencing three extra chloroplast markers for a select group of taxa, followed by separate and combined (total evidence) molecular phylogenetic analyses. This approach did increase resolution at the base of the southern African lineage, but many clades still showed poor resolution and support despite the use of more than 7 000 bases of sequence data. Resolving these clades within the southern African Oxalis phylogeny remains a challenge, and should prove a fertile field for future research. Thirdly, the ages (and thus duration of presence) of many Cape plant lineages within the CFR are of major interest, given that the CFR represents a global biodiversity hotspot. The age of the genus in the Cape is estimated by analyzing combined sequence data for all sampled taxa under both a Bayesian Relaxed Clock and a semiparametric Penalised Likelihood method, using calibration points inferred from Relaxed Clock analyses of the entire order Oxalidales, for which fossil data are available. In an attempt to account for known problems with divergence time estimation, I explored the potential bias introduced by method used, marker genome source and different calibrations on the root. The results indicate substantial variation in the age of crown southern African Oxalis over a nearly twenty million year period, varying according to source data, calibration estimate and methodology employed in the reconstruction. Despite this major variability, all average estimates are older than iv 18 million years, which agrees with a growing body of evidence that there has been a gradual accumulation of floristic diversity in the CFR, rather than a rapid, recent burst of speciation. Fourthly, as the produced phylogenies conclusively show the artificial nature of the current taxonomy, I propose a new, almost completely different classification for southern African Oxalis taxa. Although a significant improvement, this classification is considered informal due to the complete disagreement between the old and proposed new taxonomies, poor resolution in some of the proposed lineages, and a need to confirm proposed groups (clades) with the identification of morphological synapomorphies. Potential synapomorphies for various clades are proposed and discussed, which should guide future research. Fifthly, the presence of bulbs in this genus is of great interest as a potential preadaptation for seasonally arid climates. The evolution of the bulbous habit in Oxalis is here explored for the first time. I address the sequence of major morphological character state changes leading to the suite of characters corresponding to the bulbous habit. The homology of basal leaf petioles, fleshy leaf scales and tunics is discussed, and it is shown that many bulb characters present in the southern African lineage are also found in the close relatives of this lineage, and are thus older than this lineage. The ecological and evolutionary implications of bulb geophytism in the CFR Oxalis are also discussed. Finally, I address certain taxonomic issues that arose during the course of this study. Co-authors and I describe the new species O. saltusbelli and O. ericifolia. We also clarify issues surrounding the tremendously variable group species O. flava and propose some nomenclatural changes and synonyms for related taxa. We also address the taxonomic position of the rare species O. purpurata, which was located too late in the course of this study to include in the main analyses.

Thesis (PhD (Botany and Zoology))—-Stellenbosch University, 2008. / Streptocarpus Lindl. (Gesneriaceae DC.) is a genus of herbaceous plants containing approximately 160 species, of which the majority occur in Africa and Madagascar. They are largely restricted to shaded and moist habitats such as primary forest and rock outcrops. The genus contains considerable morphological variation, with subgenus Streptocarpella containing caulescent species and subgenus Streptocarpus mostly consisting of acaulescent growth forms, mainly the unifoliates, plurifoliates and rosulates. Preliminary molecular analyses conducted using nuclear ITS sequence data suggested that subgenus Streptocarpus evolved in tropical central Africa, before radiating in several independent waves into southern Africa. Streptocarpus has therefore only recently spread into South Africa. Amongst the South African species, 11 morphologically similar species were identified as being closely related, together forming the Cape primrose clade, based on the analysis of nuclear ITS sequence data. However, these analyses only contained a few South African species, and the ITS data did not provide enough resolution of relationships within this clade. In this study nuclear and chloroplast sequence data as well as nuclear microsatellite data were therefore employed to unravel the complex relationships amongst the South African Streptocarpus species. The analyses indicate that 16 rosulate, palynologically similar species (S. primulifolius, S. rexii, S. johannis, S. baudertii, S. modestus, S. formosus, S. gardenii, S. lilliputana, the S. cyaneus complex [S. cyaneus, S. parviflorus, S. fenestra-dei, S. kunhardtii and S. roseo-albus], S. floribundus, S. aylae and S. kentaniensis), the core Cape primrose species, are closely related, while five unifoliate/plurifoliate, palynologically more variable species (S. denticulatus, S. dunnii, S. pusillus, S. rimicola and S. bolusii) consistently emerged as more distantly related to the core Cape primrose species. However, the positions of a further ten species (S. meyeri, S. montigena, S. fanniniae, S. caeruleus, S. longiflorus, S. polyanthus, S. saundersii, S. porphyrostachys, S. grandis and S. vandeleurii) were more complex in the analyses, indicating that hybridization has played a role in their evolution. Five of these species (S. meyeri, S. montigena, S. fanniniae, S. caeruleus and S. longiflorus) are, however, palynologically homogenous and rosulates, and therefore are probably more closely related to the core Cape primrose species, while the other five (S. polyanthus, S. saundersii, S. porphyrostachys, S. grandis and S. vandeleurii) are unifoliates/plurifoliates that are palynologically more heterogenous, and are probably more distantly related to the core Cape primrose species. Amongst the core Cape primrose species, S. primulifolius emerged as being ancestral or having hybridized with many of the other species, while the S. cyaneus complex forms a geographically and genetically more isolated group. However, evolutionary relationships amongst these species were to a certain extent obscured by incomplete lineage sorting caused by limited interpopulation gene flow, frequent hybridization and rapid speciation. The analyses confirmed that the Pondoland Centre forests constitute important Pleistocene refugia, and revealed some of the historical migration routes along which the species had radiated.

One of the factors that affect the post-harvest behavior of seeds is provenance. Different geographical locations are characterised by different environmental conditions, hence, plant materials harvested from different locations may respond differently to laboratoryinduced stresses. The aim of the present study was to assess the role that provenance plays on the degree of recalcitrance of a single species – a factor that needs to be taken into consideration when choices are to be made for plant germplasm conservation. In this study, seeds of Trichilia dregeana, which have been shown to display recalcitrant post-harvest behavior, were harvested from four locations that were slightly different in climatic conditions: Mtunzini (MTZN), Durban (DBN), Pietermaritzburg (PMB) and Port Edward (P.ED), all within the KwaZulu-Natal province of South Africa. Clean seeds, whose aril and seed coat have been removed, were subjected to different drying and chilling stresses and their responses to the stresses are examined. Excised embryonic axes of the seeds were also subjected to flash (very rapid) drying and their responses to the stress are shown. Although the vegetation of these locations can be categorized as being sub-tropical, the present study suggests that there may be ‘degrees’ in the sub-tropical nature based on the rainfall and maximum temperature data. The ‘degree of sub-tropicality’ is suggested to be in the order MTZN > DBN > P.ED > PMB. Mean seed size assessed as seed length, width and fresh mass varied significantly across the four regions, with seeds from MTZN being the smallest and those from P.ED being the largest. The shedding water content of the embryonic axes of the seeds for 150 min. However, shoot production, root and shoot length and dry mass accumulation showed that axes of seeds from MTZN were the most adversely affected by rapid drying while those from PMB were the least sensitive. When clean whole seeds were dried slowly by burying in silica gel for 36 h, all seeds of the four provenances still retained more than 50% of their shedding water. MTZN seeds lost viability completely from the 24 h of slow drying while seeds from the other three locations retained some viability after 36 h. The order of survival of slow drying was MTZN < DBN < P.ED = PMB. Storage of cleaned whole seeds at chilling temperatures (3oC, 6oC and 16oC) caused increase in the water content of the embryonic axes of the seeds across the four locations. All seeds of the four provenances had lost more than 50% survival after they had been stored at 3oC for 10 weeks. Survival was completely lost from the 12th week for MTZN seeds, from the 14th week for DBN and P.ED seeds and from the 16th week for PMB seeds. There was no survival recorded for seeds across the four regions at the 18th week. The seeds were able to tolerate storage at 6oC for a little longer than those stored at 3oC. Only MTZN seeds completely lost viability at the 18th week; seeds from the other three locations retained some viability throughout the 18 weeks of storage. Storage at 16oC was the most tolerable, as all seeds from the four provenances maintained some viability until the 18th week in storage, with MTZN having the least survival and those from P.ED having the highest survival. Phylogenetic analysis of the internal transcribed spacer (ITS) region of the DBN and PMB samples showed intraspecific levels of genetic variation, and were separated by a genetic distance of 0.9%, an indicator that differences in storage behavior, chilling and desiccation sensitivity between seeds obtained from different collection localities may be genetically based. This study showed that MTZN seeds were the most sensitive, while seeds from PMB and P.ED were the most tolerant, to drying and chilling treatments, respectively. The prevailing climatic conditions may have a bearing on the responses observed in this study. / Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2011.

The hydraulic conductance of a plant is a significant factor in determining the / Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2008.

Genetic diversity of cultivated species and their wild relatives, as well as of wild species encompasses plant genetic resources or germplasm, the ex situ preservation of which embodies a critical aspect of biological conservation. While seed storage affords an efficient ex situ conservation method, recalcitrant seeds are intolerant of desiccation and cannot be stored conventionally in seed banks. Seeds of the three indigenous tree species investigated in this study, viz. Trichilia emetica, T. dregeana and Protorhus longifolia are recalcitrant, with the species considered to be endangered. Cryopreservation, which involves storage at ultra-low temperatures of selected tissue(s) from which plants are subsequently able to be generated, is currently the only method available for long-term ex situ conservation of recalcitrant-seeded species and affords significant potential for the future. Many protocols that have been applied for the cryopreservation of the germplasm of recalcitrant zygotic embryonic axes excised from seeds of tropical/sub-tropical species have resulted in survival post-cryo which has been recorded only as root development or callus formation, with shoot formation seldom occurring. Successful cryostorage of genetic resources cannot be achieved until post-cryopreservation recovery facilitates normal seedling development, i.e. the formation of both a fully functional root and a shoot. Cryopreservation requires the utilisation of the smallest explant possible (greatest surface area to volume ratio), the most suitable for recalcitrant seeds in general being the zygotic embryonic axis. Based on preliminary studies it was demonstrated that shoot production by axes is inhibited in association with a burst of reactive oxygen species (ROS), produced in response to wounding upon excision of the axis from the cotyledons, when these are attached close to the shoot apical meristem. It was postulated that a combination of the oxidative burst at the site of excision coupled with inadequate antioxidant machinery within the recalcitrant axis tissue, precludes shoot production. It was further considered highly probable that each subsequent stressful manipulation throughout the cryopreservation process would be accompanied by a surge of uncontrolled oxidative activity within the tissue, in response to the stress. Therefore, the primary aim of the study was to investigate the underlying causes of failure of shoot production after procedures associated with cryopreservation and to focus on ways to ameliorate the consequences of unbalanced oxidative metabolism. Additionally, studies were carried out to optimise each step of the cryopreservation procedure, viz. cryoprotection, dehydration, rehydration and cooling, and subsequent recovery, in conjunction with assessment of oxidative responses, ultimately to achieve successful cryopreservation of the embryonic axes of these species. The experimental work conducted to achieve this aim assessed changes in various biomarkers of injury, those selected for this study being three ROS, viz. superoxide, the hydroxyl radical and hydrogen peroxide, after axes were exposed to various pre-treatments, cryopreservation and recovery. Concomitantly, the elicited responses of endogenous antioxidant systems accompanying these steps were assessed. Changes in the levels of ROS and antioxidant activity were determined using various biochemical assays, and these parametres, together with assessment of shoot development, were investigated after each step of the cryopreservation process. The effect of stress on oxidative metabolism was tested after exposure to pre-treatments with and without the provision of various antioxidants, viz. DMSO, ascorbic acid and cathodic water, so as to determine the efficacy of selected ROS scavengers and, in general, to develop the best protocol for cryopreservation of embryonic axes of the three species. Significant results, in terms of shoot development and regulated ROS generation, were obtained after three major processes of the cryopreservation procedure. The production of roots and shoots by excised axes of T. emetica, T. dregeana and P. longifolia after excision (75%, 80% and 75%, respectively), and by 40% of excised axes of T. dregeana after each of the two further stages, cryoprotection and desiccation, were major achievements towards cryopreservation of the recalcitrant germplasm. The modulation of ROS by ascorbic acid and cathodic protection significantly improved survival of axes of both Trichilia species. In its entirety, the present study made significant advancements towards cryopreservation of recalcitrant germplasm and also towards understanding oxidative events associated with cryogenic processing and exposure to cryogenic conditions. This study concludes that unregulated metabolism is one of the underlying causes of failure of recalcitrant germplasm represented by zygotic axes, to survive cryopreservation. The application of antioxidants and cathodic protection during cryopreservation facilitated survival that has been previously unattainable. The outcomes of this study provide an informative platform for further optimising cryopreservation procedures for the germplasm of the species investigated, and extending the work to other recalcitrant-seeded species, especially those of tropical/sub-tropical provenances. / Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2012.

No abstract available. / Thesis (M.Sc.)-University of Durban-Westville, 1986.

Thesis (MSc (Botany and Zoology))--Stellenbosch University, 2011. / ENGLISH ABSTRACT: See full text for abstract / AFRIKAANSE OPSOMMING: sien volteks vir opsomming

The endogenous cytokinin levels in the brown alga Ecklonia maxima (OSBECK) PAPENF., and the effect of applications of the seaweed concentrate (Kelpak 66) prepared from this alga, on the growth and yield of various plants was investigated. Tentative identification of the cytokinins present in Ecklonia maxima using High Performance Liquid Chromatography revealed the presence of cis and trans-ribosylzeatin, trans-zeatin, dihydrozeatin and isopentenyladenosine. Seasonal and lunar variations in the endogenous cytokinin levels in fresh and processed Ecklonia maxima material were investigated. Lamina, stipe and holdfast regions of one, two and three metre plants harvested from February 1981 until January 1982 together with samples of processed material from the normal production run, collected over the same period were used in this investigation. Analysis revealed both qualitative and quantitative changes in the cytokinin levels which were closely correlated to the seasonal patterns of growth of Ecklonia maxima. During summer zeatin, ribosylzeatin and their dihydroderivatives were responsible for most of the detected activity. The cytokinin glucosides increased above the levels of free cytokinins during winter. The lunar cycle study of material harvested on a daily basis during April - May 1983 revealed marked fluctuations in the cytokinin levels in the various tissues of two metre plants which were closely correlated with the phases of the moon. Greenhouse trials were conducted to determine the effects of the commercially available seaweed concentrate (Kelpak 66) on the growth of Lycopersicon esculentum MILL. plants in nematode infested soil. Kelpak 66 at a dilution of 1 : 500 improved the growth of treated plants significantly, irrespective of whether it was applied as a foliar spray at regular intervals, or whether the soil in which the plants were grown was flushed once with the diluted seaweed concentrate. Root growth was significantly improved whenever the seaweed concentrate was applied. Associated with this improved root growth was a reduction in the infestation of Meloidogyne incognita (KOIFORD and WHITE) CHITWOOD. Finally, the effect of seaweed concentrate and fertilizer applications on the growth and endogenous cytokinin content of Beta vulgaris L. and Phaseolus vulgaris L. plants was investigated. Seaweed concentrate at a dilution of 1 : 500 applied as a foliar spray improved the growth of treated plants significantly, irrespective of whether it was applied on its own or together with a chemical fertilizer. Root growth and the endogenous cytokinin content of these roots increased with seaweed concentrate application. Increases were also detected in the cytokinin content of fruits of Phaseolus vulgaris plants treated with seaweed concentrate. Associated with this increase in the cytokinin content was an increase in the dry mass of the fruit from treated plants. The significance of these findings and the possible relationship between the endogenous cytokinins present in Ecklonia maxima and the effect of the seaweed concentrate on plant growth is discussed. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 1984.

Bananas are climacteric fruits and show climacteric patterns with respect to both their ethylene production and respiration rates during the ripening process. Treatment with ethylene accelerated the ripening process, evidenced by colour changes. Chlorophyllase apparently synthesised chlorophyll in the 0.2 kGy irradiated fruit during the post-climacteric period. Gamma irradiation of green bananas with doses of 0.4 kGy or greater resulted in extensive tissue damage, undesirable colour development, and drastic changes in respiration and ethylene production. Irradiation at 0.2 kGy, did however, effectively delay ripening (without causing any phytotoxicity) of the fruit apparently by decreasing its sensitivity to its own endogenous ethylene for a period up to 28 days when kept at room temperature, provided that the relative humidity was maintained at 75 % to prevent desiccation. Irradiation at this dose did not prevent the onset of ripening when subjected to high concentrations of exogenous ethylene. Ultrastructurally total cell collapse due to cell wall, middle lamella and membrane damage was evident at doses of 0.4, 0.6 and 1.0 kGy. Irradiation with a dose of 0.2 kGy did not cause cell wall softening and membrane breakdown and thus provided a banana fruit of excellent postharvest quality with respect to ultrastructural features. Glucose and fructose occurred probably directly from sucrose that may not accumulate because it is transient after the banana fruit is harvested. The accumulation of sucrose glucose and fructose was not stimulated by ethylene in non-irradiated and irradiated fruit. These results indicate a clear temporal separation between those biochemical processes that appear to be stimulated by ethylene (respiration, fruit softening and colour change) from those that are not (sugars). Pectin methyl esterase resulted in the demethylation of pectin-like material in the cell walls of ripening bananas. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 1991.