Rozprawy doktorskie na temat „Physiological and structural Botany”

Mediterranean species have to be in constant acclimation to seasonal environmental changes, including variations in water availability, air temperature and solar radiation. Due to global warming, these environmental changes are predicted to increase in frequency and intensity. Shrubs have a crucial role in plant communities such as avoiding soil erosion and, therefore, the study of shrub responses to abiotic stresses is essential to predict future changes in populations. Here, we evaluated the performance of the Mediterranean shrub, Cistus albidus, under several abiotic stresses in natural and controlled conditions. It was found that C. albidus plants have to face several combinations of abiotic stressors, with drought and cold being the main environmental cues that threaten their performance during winter. As a first line of defence, C. albidus diurnally changed the angle of their most apical leaves together with a decussate leaf disposition to avoid photoinhibition. Structural mechanisms were not always effective for the whole plant, and carotenoids had a primordial role during the day, dissipating excess energy as a second line of defence. They also had an important function under sustained winter stress, but α-tocopherol was the crucial antioxidant responding to stress, as a third line of defence. Jasmonates seemed to be involved in many abiotic stress responses, including cold stress and drought. High jasmonates contents, especially those of 12-oxo-phytodienoic acid, jasmonic acid and jasmonoyl-isoleucine, were found in natural conditions during a particularly cold day and after a winter drought event in C. albidus seedlings. Furthermore, it is shown that α-tocopherol and jasmonates have an important role in drought tolerance and survival of seedlings in natural field conditions. Even though seedlings have evolved numerous mechanisms to tolerate stress, there was a 30% decrease in survival after summer drought. It is concluded, given the variability in stress responses and the number of mechanisms to withstand abiotic stress, that C. albidus is a highly tolerant and resilient plant to abiotic stress. However, there are still tipping points that could become more frequent in the wake of global change that could cause shifts in C. albidus populations.

1. The septal structure of C. cinereus and N. crassa was examined via fluorescence microscopy and transmission electron microscopy. The septa of these organisms were shown to contain two plates of chitin, not one, as had been previously believed.2. The effect of the fluorochrome Calcofluor White on chitin synthesis in vivo and in vitro was investigated. Growth of C. cinereus in media seeded with the dye caused abnormal deposition of chitin at points of growth i.e. tips, septa, hook cells and clamps. However, high concentrations of the dye were required to have an effect on the chitin content and growth rate of the mycelium. X-ray crystallography showed that chitin synthesized in the presence of the fluorochrome was less crystalline than chitin synthesized when the dye was absent. It would appear, therefore, that Calcofluor binds to nascent chitin inhibiting its crystallization into microfibrils. It was concluded that chitin synthesis is a two-step process involving polymerization and subsequent crystallization.3. A range of fungi and growth forms were probed for the presence of actin using the actin-specific fluorochrome rhodamine phalloidin. Only U. phaseoli germlings and C. albicans yeast cells appeared to contain the protein. It is suggested that the failure to visualize actin in the other organisms examined is linked to their cell wall structure which prevented penetration of the rhodamine phalloidin to sites of actin localization.4. In the light of these results, mechanisms of hyphal growth are discussed and a model for the polarity of apical extension is presented.

The use of oats for human consumption is increasing every day due to the health benefits of oat products. With the objective to study relationships among factors affecting oat grain quality, two Recombinant Inbred Lines (RIL) mapping populations (`ND030299' x `ND991151' and `ND030299' x `Souris') have been used in this study. The two populations with their parents and three check cultivars were evaluated in a square lattice design in 2008 and 2009 at two North Dakota locations. Data were recorded on the following agronomic traits: grain yield, test weight, 1000 kernel weight, thin kernels, heading date, and plant height. Chemical and grain physical analysis were performed for ?-glucan, oil, and groat percentage. A total of 4975 SNP markers were assessed on the two populations using a 32-bead chip platform developed by Illumina. QTLs for agronomic and grain physical traits were mapped and characterized in the two populations using Windows QTL Cartographer. Grain yield was positively correlated with test weight, thin kernels, plant height, ?-glucan content, and associated negatively with 1000 kernel weight. Thirty linkage groups using 1168 polymorphic markers were formed for population 05021, whereas population 05026 comprised 33 linkage groups using 1024 polymorphic markers. The 30 linkage groups of population 05021 contained from 3 to 62 markers, and varied in size from 15.8 to 225.3 cM for a total map size of 2601.7 cM. The 33 linkage groups of population 05026 comprised from 2 to 42 markers, and varied in size from 2.3 to 143.2 cM for a total map size of 1174.2 cM. Nineteen genomic regions on 14 linkage groups were significantly associated with agronomic and grain chemical traits in the population 05021. Fourteen genomic regions on 12 linkage groups were identified for agronomic traits in the population 05026. The same genomic region on LG 05021-16 was associated with thin kernels, test weight, 1000 kernel weight, and oil content. LG 05026-19 loci, from position 23.7 to 47 cM, had strong effects on heading date, plant height, and grain yield. The QTLs consistently detected across environments and between the two populations could serve as starting points for marker-assisted selection.

Some responses of turfgrasses to trampling stress were examined using simulated trampling methods in greenhouse and laboratory studies. Subsidiary experimental field studies indicated that the laboratory results were relevant to the natural situation. Shoot yield, 002 exchange and water relation parameters were measured, concentrating on the short term response to simulated trampling. A range of turf grass species and cultivars were included so comparison of wear tolerant and wear susceptible cultivar response was possible. An electrolyte release method was developed to ascertain plant injury following simulated trampling. Differences in injury index were related to levels of wear tolerance of the species and cultivars. Shoot yield was reduced with simulated trampling in all species and cultivars tested. Decrease in shoot yield correlated significantly with increasing intensity of trampling and increased injury index. Differences in amount of shoot decrease between species and cultivars related to ratings of wear tolerance in the literature. Net photosynthesis of Lolium perenne S23 decreased significantly with simulated trampling and a slight increase in respiration was recorded. Net photosynthesis correlated with injury level. Continuous monitoring over the initial few hours after treatment revealed a sharp decline in photosynthesis rates, followed by gradual recovery. Selected cultivars had a lower shoot water content two hours after treatment, the higher the intensity of simulated trampling the lower was the water content. SEM studies indicated disruption of epicuticular wax, therefore reducing cuticular resistance to water loss. Transpiration rates were reduced with simulated trampling and calculations showed a reduced hydraulic conductivity. These findings were related to relevant observations of response of plants to other stresses, particularly wind. A model is presented suggesting how observed and hypothetical responses, both short term and long term, may contribute to the survival or death of grass following trampling.

One of the distinguishable plant developmental events is the transition from the vegetative to reproductive phase (RP) of development. This stage is preceded by the juvenile to adult transition within the vegetative phase. During the juvenile vegetative phase (JVP) plants are incompetent to initiate reproductive development and are effectively insensitive to photoperiod. With the change to the adult vegetative phase (AVP), plants attain competence to respond to floral inducers, which is required for the transition to the RP. This study exploits Antirrhinum and Arabidopsis as model systems to understand the genetic and environmental factors that regulate floral incompetence during the JVP. Determinants such as irradiance and [CO2] were found to be key modifiers of the JVP. A relationship between photosynthetic assimilate levels and vegetative phase transition was revealed by analysis of carbohydrates in plants at defined developmental stages. Experimental data suggest that carbohydrate levels may be required to reach a specific threshold before plants undergo the transition from a juvenile to an adult phase of vegetative growth. This may be necessary in order to sustain a steady supply of sugars for sufficient bulk flow from the leaves to the shoot apical meristem (SAM), via the phloem, to enable delivery of florigen, which thus renders the SAM competent to flower. Determination of the JVP in Arabidopsis mutants impaired in different genetic pathways has shown that multiple inputs influence the timing of the vegetative phase transition. Carbohydrates have been demonstrated to be involved possibly through their function as nutrients or signals or by their interaction with hormones. Physiological analysis of flowering time mutants has shown that a variety of signals act to promote and enable the juvenile to adult phase transition that involves both floral activators and repressors.

Bioclimatic models suggest that Protea species are likely to be severely affected by predicted increases in temperature and reductions in rainfall in the Western Cape. However, throughout their 400 million year history, land plants have been exposed to considerable climatic variations that have driven physiological adaptations promoting long-term resilience. A principal response of plants to increases in atmospheric CO2 concentration and changing rainfall regimes is to minimise water loss by reducing stomatal conductance (gs) and simultaneously increasing intrinsic water-use efficiency (iWUE). Plasticity in these two physiological traits may enhance the ability of plants to survive future climate change. There is considerable evidence in the literature for major changes in gs and iWUE in response to both short-term and long-term changes in environmental conditions. However, to date, the effects of decadal climate change on plant physiology are still largely uncertain. In this study, the responses of gs and iWUE to perturbations in rainfall and atmospheric CO2 are reconstructed from a chronological sequence of herbarium and modern Protea specimens. The results indicate that in the two high-altitude study species, P.cryophila (t= -2.44, df=7, p= 0.045) and P.venusta (t=3.08, df=5, p=0.027), stomatal conductance has significantly declined in response to increasing atmospheric CO2 concentrations over the past century. The low altitude species appears to maintain fixed stomatal conductance trajectories as a response to more xeric conditions. It is difficult to draw inferences about stomatal physiological plasticity from iWUE data as it is not possible to distinguish between the effects of physiological reductions in stomatal conductance and biochemical enhancement of photosynthesis. Microhabitat sensitivity in iWUE is a further factor confounding interpretation. This highlights the importance of using multiple parameters as analytical tools for assessing long-term physiological change. A key contribution of this study is that it has confirmed the value of using archival material and highlights methodologies that may aid future herbarium-based interpretations. In addition, despite some methodological limitations, the study has identified interesting trends at climate change-relevant timescales that point the way for further research to understand relative vulnerabilities and inform conservation strategies. Specifically, it suggests that future research and conservation efforts may need to be focused on species occurring at low altitudes because of their apparently more limited physiological plasticity.

Includes bibliographical references (leaves 145-171).
Drought stress is often the most limiting factor to maize and sorghum production in the semi-arid areas. This study evaluates the physiological (water relations, gas exchange characteristics, membrane leakage), biochemical (antioxidant protection mechanisms and photosynthetic pigment compositions) and seed viability and quality response of maize (cv Melkassa-2) and sorghum cv Macia) after exposure to and recovery from pre and post-flowering dehydration in plants grown in a controlled environment growth chamber under constant environmental conditions (12/12h day/night, 28-32/17 °c day/night temperature, 60-80% RH and PPFD of 1200-1400 umol m-2 S-1), at the Department of Botany, University of Cape Town.

Under terminal water deficit, the impact of stem carbohydrate remobilization has greater significance because post-anthesis assimilation is limited, and grain growth depends on translocation of carbohydrate reserves. The working hypothesis of this thesis is that increases in stem carbohydrates facilitate tolerance to terminal drought in wheat. The goals of this thesis are to examine this hypothesis using physiological and genetic tools; identify genes that are related to QTL for stem carbohydrate; work with wheat and barley breeders to integrate findings into the breeding program of the Department of Agricultural and Food Western Australia. The physiological data of three drought experiments (two years in a glasshouse and one year in the field) suggested the maximum level of stem water soluble carbohydrate (WSC) is not consistently related to grain weight, especially, under water deficit. The patterns of WSC accumulation after anthesis differed depending on variety and suggested that WSC degradation and translocation have different genetic determinants. Most of the carbohydrates in stem WSC in wheat are fructans. Because 1-FEH gene was an important gene in fructan degradation, the three copies of this gene (1-FEH w1, 1-FEH w2 and 1-FEH w3) were isolated from the respective genomes of bread wheat. In addition, the genes were mapped to chromosome locations and coincided with QTL for grain weight. The results of gene expression studies show that 1-FEH w3 had significantly higher levels in the stem and sheath which negatively corresponded to the level of stem WSC in two wheat varieties in both water-deficit and well-watered treatments. Strikingly, the 1-FEH w3 appeared to be activated by water deficit in Westonia but not in Kauz. The results suggest that stem WSC level is not, on its own, a reliable criterion to identify potential grain yield in wheat exposed to water deficit during grain filling. The expression of 1-FEH w3 may provide a better indicator when linked to instantaneous water use efficiency, osmotic potential and green leaf retention, and this requires validation in field grown plants. In view of the location of the contribution to grain filling of stem WSC, this is a potential candidate gene contributing to grain filling. The numerous differences of intron sequences of 1-FEH genes would provide more opportunities to find markers associated with the QTL. A new FEH gene was partially isolated from Chinese Spring and the sequence was closely related to 1-FEH genes. This gene, FEH w4, was mapped to 6AS using Chinese Spring deletion bin lines. The polymorphism of this gene was found between different bread varieties using PCRs and RFLPs, and this allowed the gene to be mapped to two populations of Hanxuan 10 x Lumai 14 and Cranbrook x Halberd. In the population of Hanxuan 10 x Lumai 14, it was close to SSR marker xgwm334 and wmc297 where the QTL of thousand grain weight and grain filling efficiency were located. This result indicated this gene might be another possible candidate gene for grain weight and grain filling in wheat.

Under terminal water deficit, the impact of stem carbohydrate remobilization has greater significance because post-anthesis assimilation is limited, and grain growth depends on translocation of carbohydrate reserves. The working hypothesis of this thesis is that increases in stem carbohydrates facilitate tolerance to terminal drought in wheat. The goals of this thesis are to examine this hypothesis using physiological and genetic tools; identify genes that are related to QTL for stem carbohydrate; work with wheat and barley breeders to integrate findings into the breeding program of the Department of Agricultural and Food Western Australia. The physiological data of three drought experiments (two years in a glasshouse and one year in the field) suggested the maximum level of stem water soluble carbohydrate (WSC) is not consistently related to grain weight, especially, under water deficit. The patterns of WSC accumulation after anthesis differed depending on variety and suggested that WSC degradation and translocation have different genetic determinants. Most of the carbohydrates in stem WSC in wheat are fructans. Because 1-FEH gene was an important gene in fructan degradation, the three copies of this gene (1-FEH w1, 1-FEH w2 and 1-FEH w3) were isolated from the respective genomes of bread wheat. In addition, the genes were mapped to chromosome locations and coincided with QTL for grain weight. The results of gene expression studies show that 1-FEH w3 had significantly higher levels in the stem and sheath which negatively corresponded to the level of stem WSC in two wheat varieties in both water-deficit and well-watered treatments. Strikingly, the 1-FEH w3 appeared to be activated by water deficit in Westonia but not in Kauz. The results suggest that stem WSC level is not, on its own, a reliable criterion to identify potential grain yield in wheat exposed to water deficit during grain filling. The expression of 1-FEH w3 may provide a better indicator when linked to instantaneous water use efficiency, osmotic potential and green leaf retention, and this requires validation in field grown plants. In view of the location of the contribution to grain filling of stem WSC, this is a potential candidate gene contributing to grain filling. The numerous differences of intron sequences of 1-FEH genes would provide more opportunities to find markers associated with the QTL. A new FEH gene was partially isolated from Chinese Spring and the sequence was closely related to 1-FEH genes. This gene, FEH w4, was mapped to 6AS using Chinese Spring deletion bin lines. The polymorphism of this gene was found between different bread varieties using PCRs and RFLPs, and this allowed the gene to be mapped to two populations of Hanxuan 10 x Lumai 14 and Cranbrook x Halberd. In the population of Hanxuan 10 x Lumai 14, it was close to SSR marker xgwm334 and wmc297 where the QTL of thousand grain weight and grain filling efficiency were located. This result indicated this gene might be another possible candidate gene for grain weight and grain filling in wheat.

Thesis (PhD) -- Stellenbosch University, 1991.
ENGLISH ABSTRACT: The effect of partial defoliation as canopy management practice on metabolism and grape composition of the grapevine, Vitis vinifera L cv. Cabernet Sauvignon, was investigated. The establishment of physiologically sound principles on how to overcome deleterious effects associated with vigorous and dense-canopy vines is emphasized. Experimental vines were defoliated 33 % and 66 % evenly over the whole canopy from different developmental stages in a field study. Effects on canopy microclimate, photosynthesis, photosynthate translocation, vegetative growth, reproductive growth, root development and distribution, as well as grape and wine quality, were determined. A method for the simultaneous extraction of sugars and organic acids from freeze-dried berries at different developmental stages is described. Partial defoliation of vines improved canopy microclimate and photosynthetic efficiency of remaining leaves. Normal translocation and distribution patterns of photosynthates were apparently unaffected by partial defoliation. Translocation to and accumulation of photosynthetic products in the leaves and bunches of partially defoliated vines were, however, improved. Remaining leaves of partially defoliated vines were in comparison photosynthetically more active. Apart from a less favourable canopy microclimate, it seemed that the sink capacity of non-defoliated vines did not comply to the source capacity, inducing a reduced rate of photosynthesis. Normal sigmoidal growth patterns of vines were not affected by partial defoliation as applied in this study. This is important for the longevity, healthiness and productivity of vines. Vegetative growth was differentially affected by partial defoliation. No compensatory leaf growth occurred in reaction to partial defoliation from different developmental stages. Main shoot length, however, decreased slightly. Lateral shoot length and number of laterals increased, whereas cane mass decreased when vines were partially defoliated, particularly the earlier and more severe the defoliation. Reproductive growth in terms of yield was deleteriously affected by 33 % defoliation prior to pea size and 66 % defoliation prior to veraison. Budding percentage was, however, improved by 33 % and 66 % defoliation, whereas bud fertility was only improved by 33 % defoliation. Partial defoliation changed the canopy microclimate to conditions favourable for pest and disease control and higher grape quality. Subterranean growth was favourably affected by partial defoliation, particularly when applied from pea size stage. These changes included higher root densities, development of higher numbers of fine and medium diameter roots and occurrence of higher total root numbers in all soil layers. Generally, defoliations from pea size and veraison were more efficient regarding root development than defoliations from just after bud break and from berry set. Partially defoliated vines reacted by forming new roots, creating a more efficient nutrient absorption capacity and utilization of soil and available water and that, together with higher photosynthetic activities of leaves, provided an efficient mechanism for continued high performance. Grape quality was not affected markedly by partial defoliation. Total soluble solids in berries of defoliated vines were comparable to and even significantly higher than those of non-defoliated vines in some cases, in spite of much lower leaf areas. Generally, total titratable acidity of musts was also slightly higher for partially defoliated vines. Glucose and fructose concentrations in berries were unaffected by partial defoliation, while tartaric acid concentrations were slightly increased and malic acid concentrations slightly decreased. Partial defoliation generally increased the anthocyanin concentration of berry skins. These changes in grape composition suggest higher grape quality and seemed to result from improved light conditions in the canopy interior. Berry volume decreased with partial defoliation, which lowered the pulp:skin ratio. These berries are more desirable for quality wines. Regardless of severity or the developmental stage defoliation was commenced, wine cultivar character and overall wine quality were significantly improved. Partial defoliation changed the general metabolism of vines, mainly in terms of more favourable source:sink ratios, resulting in more efficient photosynthesis, subterranean performance and canopy microclimate. In general, the results suggest that an even removal of 33 % of leaves opposite and below bunches during the period from flowering or berry set to pea size stage may be applied. lt is further suggested that existing vigorous and dense-canopy vines be 33 % defoliated evenly on the lower half of the shoot (canopy) from pea size or veraison. This hypothesis proved effective in improving canopy microclimate, photosynthetic activity and yield, while vegetative growth was inhibited. Grape and wine quality were higher. On the whole, partial defoliation as applied in this study, is recommended as canopy management practice in order to facilitate the abolishment of deleterious effects of excessive vegetative growth and canopy density on balanced metabolic activity, fruit and wine quantity and quality, as well as longevity and healthiness of grapevines.
AFRIKAANSE OPSOMMING: Die invloed van blaarverwydering as loofbestuurspraktyk op die metabolisme en druifsamestelling van die wingerdstok, Vitis vinifera L cv. Cabernet Sauvignon, is ondersoek. Die vestiging van fisiologies-betroubare beginsels oor hoe nadelige effekte geassosieerd met geil en lower-verdigte wingerde uitgeskakel kan word, word beklemtoon. Proefstokke is in 'n veldondersoek 33 % en 66 % eweredig oor die hele lower vanaf verskillende ontwikkelingstadiums ontblaar. Effekte op lowermikroklimaat, fotosintese, translokasie van fotosintetiese produkte, vegetatiewe groei, reproduktiewe groei, wortelontwikkeling en verspreiding asook druif- en wynkwaliteit is bepaal. 'n Metode vir die gelyktydige ekstraksie van suikers en organiese sure uit gevriesdroogde korrels op verskillende groeistadiums is ontwikkel. Blaarverwydering het lowermikroklimaat verbeter en fotosintetiese doeltreffendheid van oorblywende blare op die stok verhoog. Normale translokasie en verspreidingspatrone van produkte van fotosintese is skynbaar nie deur blaarverwydering befiwloed nie. Translokasie na, en akkumulering van fotosintetiese produkte in die blare en druiwe van gedeeltelik ontblaarde stokke, is egter verbeter. Oorblywende blare van gedeeltelik ontblaarde stokke was fotosinteties meer aktief. Afgesien van 'n ongunstiger lowermikroklimaat, het die sinkkapasiteit van nie-ontblaarde stokke skynbaar ook nie teen die bronkapasiteit opgeweeg nie en is 'n verlaagde tempo van fotosintese verkry. Normale sigmofdale groeipatrone van die stokke is nie deur blaarverwydering befrivloed nie. Dit is belangrik vir langlewendheid, gesondheid en produktiwiteit van stokke. Vegetatiewe groei is differensieel deur blaarverwydering befrivloed. Geen kompenserende blaargroei het in reaksie op blaarverwydering vanaf verskillende ontwikkelingstadiums voorgekom nie. Hooflootlengte was egter effens korter. Sylootlengte en aantal sylote het toegeneem, terwyl lootmassa afgeneem het met blaarverwydering, veral hoe vroeer en strawwer dit toegepas is. Reproduktiewe groei in terme van opbrengs is nadelig befrivloed deur 33 % ontblaring voor ertjiekorrelstadium en 66 % ontblaring voor die deurslaanstadium. Botpersentasie is egter deur blaarverwydering verbeter, terwyl oogvrugbaarheid slegs deur 33 % ontblaring verbeter is. Blaarverwydering het 'n gunstige lowermikroklimaat vir die beheer van plae en siektes en hoer druifkwaliteit geskep. Ondergrondse groei is gunstig deur blaarverwydering befrivloed, veral wanneer toegepas vanaf ertjiekorrelstadium. Hierdie veranderinge het ingesluit hoer worteldigtheid, ontwikkeling van groter hoeveelhede wortels met 'n fyn en medium deursnit en die voorkoms van 'n groter aantal totale wortels in alle grondlae. Blaarverwydering vanaf ertjiekorrel- en deurslaanstadia was in die algemeen meer doeltreffend ten opsigte van wortelontwikkeling as blaarverwydering vanaf net na bot en vanaf korrelset. Gedeeltelik ontblaarde stokke het gereageer deur nuwe wortels te vorm, waardeur 'n meer doeltreffende vermoe tot voedingstofopname en benutting van grand an beskikbare water verkry is. Tesame met hoer fotosintetiese aktiwiteite van die blare is 'n doeltreffende meganisme vir aanhoudende hoe prestasie verkry. Druifkwaliteit is nie aanmerklik deur blaarverwydering beirwloed nie. Totale oplosbare stowwe in druiwe van gedeeltelik ontblaarde stokke was vergelykbaar en selfs betekenisvol hoer as die van nie-ontblaarde stokke in sekere gevalle, ten spyte van die baie laer blaaroppervlakte. In die algemeen was die totale titreerbare suur in die mos van gedeeltelik ontblaarde stokke oak effens hoer. Glukose- en fruktosekonsentrasies in die druiwe is nie deur blaarverwydering beinvloed nie. Die konsentrasie wynsteensuur is egter effens verhoog en die appelsuurkonsentrasie effens verlaag. Antosianienkonsentrasie van korreldoppe is in die algemeen deur blaarverwydering verhoog. Hierdie veranderinge in druifsamestelling dui op hoer druifkwaliteit en is skynbaar die resultaat van verbeterde ligtoestande in die binnekant van die lower. Korrelvolume het afgeneem met blaarverwydering. Dit het die pulp:dop verhouding verlaag. Sulke korrels is meer gewens vir bereiding van kwaliteitswyne. Wyn cultivarkarakter en totale wynkwaliteit is betekenisvol deur blaarverwydering verhoog, onafhanklik van die strafheid of ontwikkelingstadium waarvandaan blare verwyder is. Blaarverwydering het die algemene metabolisme van die wingerdstok verander, hoofsaaklik ten opsigte van meer gunstige bron:sink-verhoudings en derhalwe meer doeltreffende fotosintese, ondergrondse groei en lowermikroklimaat. Die resultate dui in die algemeen daarop dat 'n eweredige blaarverwydering van 33 % regoor en onderkant die trosse tydens blomvorming of korrelset tot ertjiekorrelstadium toegepas kan word. Die resultate dui verder daarop dat 33 %van bestaande geil en lower-verdigte stokke se blare eweredig verwyder kan word op die onderste helfte van die loot (lower) vanaf ertjiekorrel- of deurslaanstadium. Hierdie hipotese was suksesvol in die verbetering van lowermikroklimaat, fotosintetiese aktiwiteit en oesmassa. Vegetatiewe groei is gestrem. Druif- en wynkwaliteit was hoer. In die geheel kan blaarverwydering soos toegepas in hierdie ondersoek aanbeveel word as loofbestuurspraktyk ten einde die uitskakeling van nadelige effekte van oormatige vegetatiewe groei en lowerdigtheid op gebalanseerde metaboliese aktiwiteit, druif- en wynkwantiteit en kwaliteit, asook langlewendheid en gesondheid van wingerdstokke te bevorder.

Two subspecies of Leucospermum conocarpodendron with considerable variation in leaf traits occur along the Cape Peninsula along clearly delimited geographical distributions. We attempted to quantify the difference in leaf trait dimensions between green (L. conocarpodendron ssp. viridum) and grey (L. conocarpodendron ssp. conocarpodendron) individuals. Leaves from grey individuals are shown to have higher reflectance across the entire photosynthetically active region (PAR) and greater stomatal density. Leaves from grey individuals were found to have higher specific leaf area (SLA] and a greater size boundary layer. We found no correlation between the transpiration rate and stomatal density, size of the boundary layer and hairiness. We hypothesize that the leaf traits are having an effect on rate of photosynthesis and subsequently determining growth strategy of each type. Grey individuals, because of increased reflectance from the leaves are able to persist throughout summer in hot, dry conditions, although they may suffer the cost of reduced photosynthetic rate during late winter and early summer when conditions are relatively mild. Green individuals are able to start growing earlier due to higher SLA and increased rate of photosynthesis but are not able to persist for as long as the grey individuals during mid to late summer. Finally we argue that adaptation to edaphic environment may be associated with subsequent shifts in flowering phenology.

Plants, like most organisms, have developed elaborate mechanisms for anticipating periodic environmental changes. The circadian clock allows an organism to adapt its metabolic, developmental and physiological processes to coincide with favourable environmental conditions. At the centre of the Arabidopsis thaliana clock, linking environmental inputs and driving the overt biological rhythm is a central oscillator that consists of multiple interlocked transcriptional/translational negative feedback loops. What is known about the structure of the central oscillator comes primarily from genetic analysis. Less clear, is how putative oscillator proteins perform their perceived functions in circadian rhythm maintenance. Described are the cloning, expression and purification of clock-associated proteins; TOCI-PRR, ELF4, LUX and LIPl. Purified ELF4 was subjected to unsuccessful crystallisation trials, probably due to its intrinsically unstructured nature. A truncated form of LIP 1 was shown to be an active OTPase, representing the first example of an active OTPase in the plant clock. In addition, a protocol for the production of ssDNA aptamers has been developed (against SRRl), which can be used to replace antibody-based experimentation. The work presented discusses the difficulty in obtaining the novel, plant-specific proteins in quantities required for crystallisation, and suggests alternative methods for structural and biochemical analysis of these proteins. Moreover, this thesis combines experimental data with a range of Bioinformatic tools to aid design for subsequent biochemical expression, purification and crystallisation trials.

The $\beta$-lactamase producing plasmids of Neisseria gonorrhoeae pJD4 (Asia-type plasmid), pJD5 (Africa-type plasmid), pJD7 (Toronto-type plasmid), pGO4717 (Rio-type plasmid) and pGF1 (Nimes plasmid) are structurally related. The primary sequence of pJD4, the primary sequence around the deletion of pJD5, pJD7 and the Rio plasmid, and the insertion of pGF1 were determined using the dideoxy termination method. This is the first report which identified the exact size and location of deletion of the $\beta$-lactamase plasmids of N. gonorrhoeae based on DNA sequence analyses. The primary sequence of pJD4 was used as reference for all other plasmids. Coordinates were numbered from the PstI site. In addition, regions around the deletion of four other Toronto-type plasmids pGC1213, pGC4538, pGC5221, pGC5228, were also sequenced. Based on the DNA sequence of pJD4, a comprehensive restriction endonuclease map ot PJD4 was constructed and compared to previously published maps. Analysis of the primary sequence of pJD4 including the region homologous to TnA shows at least 62 direct repeats (DR) and 24 inverted repeats greater or equal to 10 base pairs (bp) in length. (Abstract shortened by UMI.)

Numerous "Fynbos" species of the Cape Floristic Region (CFR) have particularly fine, narrow leaves. The rates of transpiration and heat loss are partially dependent on boundary layer conductance, which is determined by leaf shape and size, surface modifications and wind speed. We expected fine-leaved species with higher boundary layer conductance to transpire faster than broad-leaved species at low temperatures whereas at higher temperatures we expected transpiration to be limited by stomata! conductance. In contrast, the rate of heat loss may be constrained by thick boundary layers in larger leaves at high temperatures. Leaf gas exchange characteristics at various temperatures were correlated with boundary layer thickness, leaf area and specific leaf area for 14 Proteaceae species using phylogenetically independent contrast species. When the temperatures of individual leaves were altered, while ambient temperature was kept at l 8°C, water loss decreased significantly at both 12°C and 30°C with increased leaf size and thus boundary layer thickness. At 30°C, small leaves with thin boundary layers resulted in leaf temperatures below ambient, while larger leaves with thicker boundary layers had leaf temperatures closer to ambient. However, at 30°C the variation in leaf temperature between the smallest and largest leaves was only 3.4°C. Such a small variation in leaf temperature is unlikely to alter temperature-dependent physiological processes. We conclude that the small boundary layer associated with small leaves enables fine-leaved species to transpire at faster rates when water is plentiful. This may be a particularly important strategy for plants that take up most of their nutrients in the wet winter months from nutrient-poor highly leached soils of the CFR region. We suggest that fine leaves are an adaptation for nutrient uptake during winter, although they may also have the benefit of improved coupling of leaf to ambient temperature during the summer drought period.

Higher temperatures under climate change are likely to result in greater evaporation and increased soil moisture deficits. Increased drought will likely cause a shift in the vegetation distribution. This study focused on woody plants of the Southeastern United States mixed forest. Although water is not a primary limiting factor in this forest, species distribution suggests that water availability exerts a strong control on the success of woody plants. I used a comparative approach to understand environmental factors and corresponding species traits that determine species composition across a gradient of water availability. I compared hydraulic architecture, vulnerability to cavitation, and the ability of xeric and mesic species to germinate, grow and survive under varying levels of water stress. Congeneric pairs composed of one xeric and one mesic species were used. Seeds were subjected to polyethylene glycol solutions of different water potentials to compare the effects of water availability on germination of xeric and mesic species. I used understory saplings to compare the difference in the xylem hydraulic properties between xeric and mesic species and the air-injection method was used to determine differences in their xylem vulnerability to cavitation. I performed a dry-down experiment to compare the ability of the xeric and mesic species to survive extreme drought by evaluating survival after re-watering subsequent to drought exposure. Growth performance was evaluated by measuring electron transport rate (ETR), stomatal conductance (gs), shoot and root biomass of seedlings that were exposed to different levels of water (well-watered, dry, and flooded) and nutrients (high and low). Results indicate that the ability to germinate under drought did not differ consistently between xeric and mesic species, but, germination ability under drought was associated with the ability to maintain turgor. Stems of xeric species were less vulnerable to wilting than mesic species under drought stress. I found greater resistance to xylem cavitation in xeric species than mesic species. It appears that cavitation resistance was independent of specific conductivity and wood density since I did not find any trade-off between specific conductivity and vulnerability to cavitation among the woody plants examined. In the growth performance study, xeric species tend to have lower whole plant biomass, higher Root:Shoot (R/S) ratio, higher coarse root mass ratio (CRMR) and less reduction in gs under drought stress than mesic species. The higher stomatal conductance and lower photorespiration rates among the mesic species may have influenced their increased photosynthetic rates, thereby producing a greater total plant biomass than xeric species. Species distribution along water availability gradients appears to be better explained by the ability of seedlings to resist cavitation, to tolerate and survive water stress, and by their biomass allocation patterns, rather than by their ability to germinate under drought.

Over the past years fruit and vegetable industry has become interested in the application of both osmotic dehydration and vacuum impregnation as mild technologies because of their low temperature and energy requirements. Osmotic dehydration is a partial dewatering process by immersion of cellular tissue in hypertonic solution. The diffusion of water from the vegetable tissue to the solution is usually accompanied by the simultaneous solutes counter-diffusion into the tissue. Vacuum impregnation is a unit operation in which porous products are immersed in a solution and subjected to a two-steps pressure change. The first step (vacuum increase) consists of the reduction of the pressure in a solid-liquid system and the gas in the product pores is expanded, partially flowing out. When the atmospheric pressure is restored (second step), the residual gas in the pores compresses and the external liquid flows into the pores. This unit operation allows introducing specific solutes in the tissue, e.g. antioxidants, pH regulators, preservatives, cryoprotectancts. Fruit and vegetable interact dynamically with the environment and the present study attempts to enhance our understanding on the structural, physico-chemical and metabolic changes of plant tissues upon the application of technological processes (osmotic dehydration and vacuum impregnation), by following a multianalytical approach. Macro (low-frequency nuclear magnetic resonance), micro (light microscopy) and ultrastructural (transmission electron microscopy) measurements combined with textural and differential scanning calorimetry analysis allowed evaluating the effects of individual osmotic dehydration or vacuum impregnation processes on (i) the interaction between air and liquid in real plant tissues, (ii) the plant tissue water state and (iii) the cell compartments. Isothermal calorimetry, respiration and photosynthesis determinations led to investigate the metabolic changes upon the application of osmotic dehydration or vacuum impregnation. The proposed multianalytical approach should enable both better designs of processing technologies and estimations of their effects on tissue.
Negli ultimi anni l'industria di trasformazione al minimo ha mostrato un crescente interesse verso i trattamenti di disidratazione osmotica e di impregnazione sottovuoto per le loro caratteristiche basse temperature di processo e per le relativamente contenute esigenze energetiche. La disidratazione osmotica, che consiste nell'immersione di tessuti vegetali in soluzioni ipertoniche, consente all’acqua presente nei tessuti di diffondere nella soluzione osmotica ed ai soluti in soluzione di diffondere, in direzione opposta, all'interno dei tessuti. L'impregnazione sottovuoto prevede l’immersione del tessuto vegetale in una soluzione di processo e consiste di due fasi successive. Durante la prima fase, la riduzione della pressione agente sul sistema solido-liquido provoca l'espansione ed il parziale rilascio nella soluzione del gas contenuto nei pori del tessuto. La seconda fase di ripristino della pressione atmosferica determina l’espansione del gas residuo nel tessuto con conseguente richiamo della soluzione esterna all'interno dei pori. L’impregnazione sottovuoto rappresenta un’interessante operazione tecnologica poiché può permette l’introduzione nei tessuti di specifiche molecole quali antiossidanti, regolatori di pH, stabilizzanti o crioprotettori. Il presente studio si è proposto di valutare, seguendo un approccio multianalitico di indagine, le principali modificazioni a carico di tessuti vegetali assoggettati a trattamenti di disidratazione osmotica o impregnazione sottovuoto. Misurazioni di tipo macro- (risonanza magnetica nucleare), micro- (microscopia ottica) ed ultrastrutturali (microscopia elettronica a trasmissione) sono state affiancate ad analisi di texture e di calorimetria a scansione differenziale. Sono stati valutati i principali effetti sulle interazioni aria-liquido in reali condizioni, sullo stato dell'acqua del tessuto e sui compartimenti cellulari. Misurazioni di calorimetria in isoterma e determinazioni dell'attività respiratoria e fotosintetica hanno infine permesso un'indagine dei cambiamenti metabolici. Tale approccio multianalitico, permettendo una valutazione complessiva delle modificazioni a carico della materia prima, può essere applicato nell’ottimizzazione dei parametri di processo sulla base delle caratteristiche ricercate nel prodotto finito.

Over the past years fruit and vegetable industry has become interested in the application of both osmotic dehydration and vacuum impregnation as mild technologies because of their low temperature and energy requirements. Osmotic dehydration is a partial dewatering process by immersion of cellular tissue in hypertonic solution. The diffusion of water from the vegetable tissue to the solution is usually accompanied by the simultaneous solutes counter-diffusion into the tissue. Vacuum impregnation is a unit operation in which porous products are immersed in a solution and subjected to a two-steps pressure change. The first step (vacuum increase) consists of the reduction of the pressure in a solid-liquid system and the gas in the product pores is expanded, partially flowing out. When the atmospheric pressure is restored (second step), the residual gas in the pores compresses and the external liquid flows into the pores. This unit operation allows introducing specific solutes in the tissue, e.g. antioxidants, pH regulators, preservatives, cryoprotectancts. Fruit and vegetable interact dynamically with the environment and the present study attempts to enhance our understanding on the structural, physico-chemical and metabolic changes of plant tissues upon the application of technological processes (osmotic dehydration and vacuum impregnation), by following a multianalytical approach. Macro (low-frequency nuclear magnetic resonance), micro (light microscopy) and ultrastructural (transmission electron microscopy) measurements combined with textural and differential scanning calorimetry analysis allowed evaluating the effects of individual osmotic dehydration or vacuum impregnation processes on (i) the interaction between air and liquid in real plant tissues, (ii) the plant tissue water state and (iii) the cell compartments. Isothermal calorimetry, respiration and photosynthesis determinations led to investigate the metabolic changes upon the application of osmotic dehydration or vacuum impregnation. The proposed multianalytical approach should enable both better designs of processing technologies and estimations of their effects on tissue.
Negli ultimi anni l'industria di trasformazione al minimo ha mostrato un crescente interesse verso i trattamenti di disidratazione osmotica e di impregnazione sottovuoto per le loro caratteristiche basse temperature di processo e per le relativamente contenute esigenze energetiche. La disidratazione osmotica, che consiste nell'immersione di tessuti vegetali in soluzioni ipertoniche, consente all’acqua presente nei tessuti di diffondere nella soluzione osmotica ed ai soluti in soluzione di diffondere, in direzione opposta, all'interno dei tessuti. L'impregnazione sottovuoto prevede l’immersione del tessuto vegetale in una soluzione di processo e consiste di due fasi successive. Durante la prima fase, la riduzione della pressione agente sul sistema solido-liquido provoca l'espansione ed il parziale rilascio nella soluzione del gas contenuto nei pori del tessuto. La seconda fase di ripristino della pressione atmosferica determina l’espansione del gas residuo nel tessuto con conseguente richiamo della soluzione esterna all'interno dei pori. L’impregnazione sottovuoto rappresenta un’interessante operazione tecnologica poiché può permette l’introduzione nei tessuti di specifiche molecole quali antiossidanti, regolatori di pH, stabilizzanti o crioprotettori. Il presente studio si è proposto di valutare, seguendo un approccio multianalitico di indagine, le principali modificazioni a carico di tessuti vegetali assoggettati a trattamenti di disidratazione osmotica o impregnazione sottovuoto. Misurazioni di tipo macro- (risonanza magnetica nucleare), micro- (microscopia ottica) ed ultrastrutturali (microscopia elettronica a trasmissione) sono state affiancate ad analisi di texture e di calorimetria a scansione differenziale. Sono stati valutati i principali effetti sulle interazioni aria-liquido in reali condizioni, sullo stato dell'acqua del tessuto e sui compartimenti cellulari. Misurazioni di calorimetria in isoterma e determinazioni dell'attività respiratoria e fotosintetica hanno infine permesso un'indagine dei cambiamenti metabolici. Tale approccio multianalitico, permettendo una valutazione complessiva delle modificazioni a carico della materia prima, può essere applicato nell’ottimizzazione dei parametri di processo sulla base delle caratteristiche ricercate nel prodotto finito.

Calcium (Ca2+) signalling regulates many neuronal functions including neurotransmission, axonal growth and development. Neuronal calcium sensor-1 (NCS-1) has been shown to be involved in many of these processes. On Ca2+ binding, NCS-1 changes conformation and exposes a hydrophobic binding pocket. In yeast, NCS-1 binds to a PI4-kinase orthologue required for survival. In mammalian cells, NCS-1 is localised to the Golgi and plasma membranes and has been linked to multiple target proteins that have roles in neuronal signalling. NCS-1 has been shown to regulate the P/Q Ca2+ channel subunit Cav2.1; although no direct binding interaction has been identified between the proteins. The Cav2.1 C-terminal tail contains two Ca2+-sensor binding regions, the IQ domain and the calmodulin (CaM) binding domain (CBD). The first part of this study investigated NCS-1 or CaM and Cav2.1 interactions using biochemical and biophysical interactions. Pull-down analysis found that NCS-1 binds to a Cav2.1 C-terminal peptide in a Ca2+-dependent manner. Use of nuclear magnetic resonance spectroscopy also showed that the IQ domain of Cav2.1 bound to NCS-1 in the presence of Ca2+, though the NCS-1 region involved in this interaction could not be identified. The second part of this study investigated NCS-1 in the model organism C. elegans. In the worm, NCS-1 is expressed predominantly in sensory neurons. An ncs-1 null mutant worm strain (XA406) was previously shown to be defective in isothermal tracking and this was linked to a requirement for NCS-1 in memory and learning. To ensure this behaviour was not caused by a locomotion or neurotransmission phenotype, these behaviours were quantified and compared to the wild-type strain. No effect of the ncs-1 null mutation was found in a quantitative body-bend assay or in an assay of aldicarb resistance. The temperature-linked behaviour was further characterised using an acute assay for temperature-dependent locomotion (TDL). In this assay, the rate of locomotion of wild-type worms decreased when the temperature was elevated from 20oC to 28oC. In contrast, the rate of locomotion of the ncs-1 null worm was significantly increased at the higher temperature. This distinct phenotype was exploited to quantify the rescue of the null strain by expression of wild-type NCS-1 and to identify potential mechanisms involved in NCS-1 function. It was established that NCS-1 regulated TDL when expressed in AIY neurons. Using information from previous studies, key structural elements of NCS-1 were investigated by expressing NCS-1 with specific point mutations or deletions. N-terminal myristoylation of NCS-1 was not functionally required. In contrast, the N- and C-terminal clefts of the hydrophobic pocket of NCS-1 were shown to be physiologically important while the C-terminal tail was not essential for function in the TDL assay. These findings allowed discrimination between two potential modes of interaction of NCS-1 with its target proteins in a physiological context.

Plants synthesize and store oil, mostly triacylglycerol (TAG), in various storage tissues that serves as a source of carbon and energy. The process is transcriptionally controlled by WRINKLED1 (WRI1), a member of the APETALA2 (AP2) class of transcription factors, that regulates most of the fatty acid biosynthesis genes. Among the four Arabidopsis WRI1 paralogs, only WRI2 is nonfunctional and failed to complement wri1-1 mutant seeds. The oleaginous Avocado (Persea americana) fruit mesocarp (60-70% DW oil) showed high expression levels for orthologs of WRI2, along with WRI1 and WRI3. While the role of WRI1 as a master seed oil biosynthesis regulator is well-established, the function of WRI1 paralogs in non-seed tissues is poorly understood. We conducted structural analyses to elucidate distinct features of avocado WRI paralogs compared to their orthologs in seed tissues. Comprehensive comparative in silico analyses of WRI1 paralogs from Arabidopsis (dicot), maize (monocot), and avocado revealed distinct features associated with their function. Our analysis showed the presence of only one AP2 domain in all WRI2 orthologs, compared to two AP2 in others. The highly conserved N-terminal region and the less conserved C-terminal regions make up the primary structure of the proteins, with amino acid composition bias characteristic of intrinsically disordered proteins (IDPs). Additionally, the avocado WRI2 showed a high proportion of random coil secondary structure, although it lacks a C-terminal intrinsically disordered region (IDR). Also, both WRI1 and WRI2 have distinct predicted phosphorylation target sites compared to their orthologs, whereas WRI2 lacks a PEST motif. Finally, through transient expression assays, we demonstrated that both avocado WRI1 and WRI2 are functional and drive TAG accumulation in Nicotiana benthamiana leaves. Our study showed that avocado WRI2 is structurally different and is functional, unlike its ortholog in Arabidopsis. This study provides us with new targets to enhance oil biosynthesis in plants.

Self-esteem is defined as: i Previous research has linked self-esteem to many factors (life-satisfaction, health, stress); however its origins, manifestations, and thus neural correlates are largely unexplored.
This thesis includes two independent neuroimaging studies investigating central nervous system variables possibly linked to self-esteem. The first study investigated hippocampal volume variations in relation to parental bonding and self-esteem. A trend was observed between the left hippocampus and retrospective mother care (smaller volumes = lesser care). The second study used fMRI to explore the differential neural correlates of high/low self-esteem when performing a social acceptance or rejection task. Here, differences in neural reward-pathways could be revealed between high and low self-esteem, suggesting an impact of self-esteem on the neural processing of social evaluation.
In conclusion, these studies provide evidence that self-esteem has strong neural correlates. This adds to previous research showing that this is a trait that is determined early in life and is self-perpetuating.

Thesis (PhD)--Stellenbosch University, 2004.
ENGLISH ABSTRACT:Six experiments, five in the glasshouse and one in the field near Nelspruit, were carried out to ascertain the effect of factors related to shoot die-back, and of water treatments on the growth and physiological responses of Pterocarpus angolensis seedlings. The study was undertaken to broaden the knowledge and understanding of the phenomenon of shoot die-back in order to enhance our ability to regenerate the species. The following experiments were carried out (a) Assessment of biomass accumulation; anatomical characteristics of the shoot apical meristem; foliar, stem and root concentration of micro- and macronutrients associated with each phenophase, (b) Effect of seedling age and seed source on the occurrence of shoot die-back under field conditions; (c) Water treatment effects on ChI. afluorescence traits of£'. angolensis seedlings obtained by assessing the fluorescence yield of photosynthetic samples subjected to dark- and light-adaptation; (d) Genetic variation in shoot die-back and other traits of sixteen halfsib families of £.. angolensis from Malawi, Namibia and Zambia grown over two die-back seasons. Experiments conducted revealed the following 1. Patterns of growth observed in phenophases are indicators of seasonal changes in annual biomass allocation to the shoot and root. Phenophases such as leaf loss and stem senescence, whether shoot die-back occurs completely or not, are directly related to the decline in above-ground biomass and declined rate of increase in root biomass respectively. Leaf flush, expansion and maturation result in increased biomass accumulation whilst shoot die-back has a minimal downregulatory effect on root biomass accumulation compared to the shoot. Shoot dieback is not sudden, therefore from the first day of germination, seedlings synchronise growth and development with the occurrence of shoot die-back. 2. Phenophasic concentration of foliar N, Ca and Mg, stem Fe and Cu and root concentrations of P, K, Mg, Fe and B are associated with shoot die-back. Patterns of mineral nutrient concentration obtained in foliage and roots but to a lesser extent in the stem, may be related to nutrient remobilisation during shoot dieback. Higher relative mineral nutrient changes during leaf yellowing and shoot die-back may be an indication of the removal of significant volumes of mobile nutrients from senescing tissues. 3. The volume of the shoot apex of E. angolensis remains constant during different phenophases which points to seasonal uniformity in the size of the apical dome. Changes in phenology associated with declined growth, or shoot die-back, is revealed through declined cell number in the tunica which is a reflection of declined mitotic activity. 4. Shoot die-back occurs in all seedlings from nursery stock planted under field conditions and all seedlings of up to two years experience complete shoot dieback. Shoot die-back takes place irrespective of seed source or the age of nursery stock that is planted. Survival after the first shoot die-back is normally low. Water treatments had no significant effect on the function of PSIJ reaction centres of P. angolensis nursery seedlings. In the case of both dark- and light-adapted leaves, water treatment had no significant effect on the measured Chi. a fluorescence parameters or the calculated parameters (specific activities, phenomenological fluxes, structure-function and performance indexes and drivingforces). 5. Water treatments affect the shape of ChI. a fluorescence transients of lightadapted compared to that of dark-adapted photosynthetic samples of E. angolensis. No significant water treatment effect was obtained for extracted and technical Chi. afluorescence parameters, specific fluxes, quantum efficiencies and phenomenological fluxes. Quantum yield, relative electron transport and quantum yield limitation, de-excitation rate constants, structure-function, performance indexes and driving forces were also not significantly different across water treatments.6. Genetic variation was observed to exist among 16 halfsib families from Malawi, Namibia and Zambia. High heritabilities were obtained for shoot die-back and other traits, indicating that shoot die-back is genetically controlled. The trait is passed from parents to offspring and it is highly probable that it occurs, throughout its natural range, in all seedlings. Since shoot die-back is genetically programmed, it remains crucial to the ability of a seedling to regenerate in the following rainy season
AFRIKAANSE OPSOMMING:Ses eksperimente, vyf in die glashuis en een in die veld naby Nelspruit, is uitgevoer om die effek van faktore wat verwant is aan die terugsterwing van lote op saailinge, sowel as om die effek van waterstres op die groei en fisiologiese responsies van Pterocarpus angolensis saailinge, te ondersoek. Die studie is ondemeem om die kennis en begrip aangaande die regenerasie-dinamika van die spesies te verbeter. Die volgende eksperimente is uitgevoer: (a) Evaluering van die effek van jisiologiese veranderings op biomassa; blaar-, stam- en wortelkonsentrasies van spoor- en makro-voedingselemente, en anatomiese eienskappe van die apikale meristeem van die lote. (b) Effek van saailingouderdom en saadbron op die voorkoms van lootterugsterwing onder veldtoestande. (c) Waterbehandelingseffekte op Chi. ajluorisensie eienskappe van ,e. angolensis saailinge wat verkry is deur die jluorisensie te evalueer van fotosintesemonsters wat aan donker- en lig-adaptasies onderwerp is. (d) Genetiese variasie in loot-terugsterwing en ander groei-eienskappe van 16 halfsib families van ,e. angolensis vanaf Malawi, Namibia en Zambia wat gekweek is oor twee terugsterj-seisoene. Die eksperimente het die volgende aan die lig gebring: 1. Groeipatrone waargeneem gedurende die fenofases is indikatore van seisoenale veranderings in jaarlikse biomassa allokasies aan die loot en die wortels. Fenofases soos blaarverlies en lootafsterwing, ongeag of loot-terugsterwing volledig is of nie, is direk verwant aan die afname in bogrondse biomassa en afnemende tempo van toename in wortelbiomassa respektiewelik. Bottende blare, vergroting en rypwording van blare lei tot toenemende biomassa akkumulasie terwyl loot-terugsterwing 'n minimale afskalende effek op akkumulasie van wortelbiomassa het in vergelyking met die van die loot. Loot-terugsterwing is nie skielik, met ander woorde vanaf die eerste dag van ontkieming sinchroniseer saailinge groei en ontwikkeling met die voorkoms van loot-terugsterwing. 2. Fenofase konsentrasies van en veranderings in blaar N en Ca en loot Fe, asook veranderings in waargenome wortel N, K, Ca, Mn, Cu, Zn en B is sterk geassosieer met loot-terugsterwing. Patrone van minerale voedingselementkonsentrasies wat in blare en wortels, en in minder mate in die loot, verkry is, mag direk verwant wees aan hermobilisering van voedingselemente gedurende loot-terugsterwing. Hoe relatiewe minerale voedingselementveranderings gedurende die vergeling van blare en lootterugsterwing mag 'n indikasie wees van die verwydering van betekenisvoUe hoeveelhede mobiele nutriente vanaf sterwende weefsel. 3. Die volume van die groeipunt van r. angolensis bly konstant gedurende verskillende fenofases wat dui op seisoenale uniformiteit in die grootte van die apikale koepel. Veranderings in fenologie ge-assosieer met afnemende groei, of loot-terugsterwing, word gerejlekteer deur afnemende selgetaUe in die tunika wat dui op afnemende mitotiese aktiwiteit. 4. VoUedige loot-terugsterwing kom voor in aUe saailinge vanaf die kwekery wat in die veld geplant word tot op die ouderdom van twee iaar. Dit kom voor angeag van saadbron of ouderdom van saailinge ten tye van planting. Oorlewing na aanvanklike loot-terugsterwing is normaalweg laag. 5. Water behandelings het geen beduidende effek op die funksie van PSII reaksiesentra van r. angolensis kewekery-saailinge gehad. Vir beide donker- en lig-aangepaste blare is geen beduidende waterbehandelingseffek verkry vir waargenome ChI. a jluoresensie parameters of die berekende parameters (spes ifieke aktiwiteite, jenomenologiese jlukse, struktuur-funksie-indekse, "perjormance-indekse oj" driving forces" ). 6. Genetiese variasie tussen 16 halfsibfamilies vanaf Malawi, Namibie en Zambie is verkry vir loot-terugsterwing en ander groei-eienskappe. Dit dui op genetiese beheer van terugsterwing en dat die eienskap oorerjbaar is, en waarskynlik in die hele natuurlike verspreidingsgebied van die spesies in aUe saailinge voorkom. Aangesien loot-terugsterwing gene ties geprogrammeer is, is dit noodsaaklik vir die vermoe van die plant om in die volgende reenseisoen te regenereer.

The complete role of inositol signaling in plants and humans is still elusive. The plant Arabidopsis thaliana contains fifteen predicted inositol polyphosphate 5- phosphatases (5PTases, E.C. 3.1.3.36) that have the potential to remove a 5-phosphate from various inositol second messenger substrates. To examine the substrate specificity of one of these Arabidopsis thaliana 5PTases (At5PTases), recombinant At5PTase1 was obtained from a Drosophila melanogaster expression system and analyzed biochemically. This analysis revealed that At5PTase1 has the ability to catalyze the hydrolysis of four potential inositol second messenger substrates. To determine whether At5PTase1 can be used to alter the signal transduction pathway of the major drought-sensing hormone abscisic acid (ABA), plants ectopically expressing At5PTase1 under the control of a constitutive promoter were characterized. This characterization revealed that plants ectopically expressing At5PTase1 had an altered response to ABA. These plants have stomata that are insensitive to ABA, and have lower basal and ABA-induced inositol (1,4,5)-trisphosphate [Ins(1,4,5)P₃] levels. In addition, At5PTase1 mRNA and protein levels are transiently regulated by ABA. These data strongly suggest that At5PTase1 can act as a signal terminator of ABA signal transduction. Like the Arabidopsis At5PTase1, a human 5PTase, Ocrl, has the ability to catalyze the hydrolysis of a 5-phosphate from several inositol-containing substrates. The loss of functional Ocrl protein results in a rare genetic disorder known as Lowe oculocerebrorenal syndrome. To gather information concerning the specificity determinants of the Ocrl protein, a structure-function analysis of Ocrl was conducted using a vibrational technique, difference Fourier transform infrared (FT-IR) spectroscopy. Upon the introduction of Ins(1,4,5)P₃ substrate, structural changes in carboxylic acid and histidine residues were observed. The net result of changes in these residues indicates that upon Ins(1,4,5)P₃ introduction, a carboxylic acid-containing residue is protonated, and a histidine residue is deprotonated. This interpretation supports the idea that the deprotonation of the histidine residue is concomitant with the coordination of a divalent cation upon Ins(1,4,5)P₃ introduction. This work allows for the proposal of a new model for the role of the active site histidine of OCRL.
Ph. D.

Pepper plants (Capsicum annuum L.) were grown in field and a greenhouse in Arizona to determine the effects of water stress, root temperature, and exogenously applied growth regulators on cytokinin production and the resulting growth. Research showed that vegetative plants were significantly higher in cytokinin activity and growth parameters than fruiting plants. Also, in root-pruned fruiting plants, cytokinin activity was less than that of intact fruiting plants. In vegetative plants, the competition between removed sinks and the rest of the shoot was reduced and, hence, more cytokinin came from the roots to the shoots. Besides, additional carbohydrates were available and recycled to the roots. In respect to temperature effect, elevating temperature from 15 to 30°C had a pronounced effect of increasing the growth rate and cytokinin activity. The measured parameters declined when temperature exceeded 30°C. Temperatures between 25 and 30°C were found to be optimum. Under experimental conditions, growth regulators (Cytex® and Burst®) applied to the soil or foliage had no significant effect on growth rates or cytokinin activity in roots. Also, applying Burst® or kinetin to the nutrient medium had inconsistent and statistically nonsignificant effects on photosynthesis and transpiration.

WRINKLED 1(WRI1), a member of AP2/EREBP class of transcription factors regulates carbon allocation between glycolytic and fatty acid biosynthetic pathway. Additionally, among the four WRI1 paralogs in arabidopsis, WRI3 and 4 but not WRI2, are also able to increase fatty acid content in different tissues. While the role of WRI1 is well established in seeds, the potential or WRI1 or its paralogs as master regulators in oil-rich nonseed tissues is poorly understood. Recent transcriptome studies of avocado (Persea americana) mesocarp revealed that the ortholog of WRI2, along with WRI1 and WRI3 was highly expressed during oil accumulation. Through transient expression assays, we further demonstrated that both PaWRI1 and PaWRI2 can accumulate oil in tobacco leaves. We conducted a comprehensive and comparative in silico analysis of WRI paralogs from a dicot, monocot and a basal angiosperm to identify distinct features associated with function. These data provide insights into the possible evolutionary changes in WRI1 homologs and allow for identification of new targets to enhance oil biosynthesis in diverse tissues.

Thesis (MSc)--Stellenbosch University, 2000.
ENGLISH ABSTRACT: This investigation was undertaken to determine whether elevated concentrations of dissolved inorganic carbon (DIC) supplied to plant roots could improve plant growth and alleviate the effects of salinity stress on tomato plants infected with arbuscular mycorrhizae. Lycopersicon esculentum cv. FI44 seedlings were grown in hydroponic culture (pH 5.8) with 0 and 75 mM NaCI and with or without infection with the fungus Glomus mosseae. The root solution was aerated with ambient CO2 (360 ppm) or elevated CO2 ( 5 000 ppm) concentrations. The arbuscular and hypha I components of mycorrhizal infection as well as the percentages total infection were decreased or increased according to the variation in seasons. The plant dry weight of mycorrhizal plants was increased by 30% compared to non-mycorrhizal plants at elevated concentrations of CO2, while the dry weight was decreased by 68% at ambient CO2 concentrations. Elevated CO2 also stimulated the growth of the mycorrhizal fungus. Elevated CO2 increased the plant dry weight and stimulated fungal growth of mycorrhizal plants possibly by the provision of carbon due to the incorporation of HCO)- by PEPc. Plant roots supplied with elevated concentrations of CO2 had a decreased CO2 release rate compared to roots at ambient CO2. This decrease in CO2 release rate at elevated CO2 was due to the increased incorporation of HC03- by PEPc activity. Under conditions of salinity stress plants had a higher ratio of N03-: reduced N in the xylem sap compared to plants supplied with 0 mM NaCI. Under salinity stress conditions, more N03- was transported in the xylem stream possibly because of the production of more organic acids instead of amino acids due to low P conditions under which the plants were grown. The N03· uptake rate of plants increased at elevated concentrations of CO2 in the absence of salinity because the HCO)- could be used for the production of amino acids. In the presence of salinity, carbon was possibly used for the production of organic acids that diverted carbon away from the synthesis of amino acids. It was concluded that mycorrhizas were beneficial for plant growth under conditions of salinity stress provided that there was an additional source of carbon. Arbuscular mycorrhizal infection did not improve the nutrient uptake of hydroponically grown plants.
AFRIKAANSE OPSOMMING: In hierdie studie was die effek van verhoogde konsentrasies opgeloste anorganiese koolstof wat aan plant wortels verskaf is, getoets om te bepaal of dit die groei van plante kan verbeter asook of sout stres verlig kon word in tamatie plante wat met arbuskulêre mikorrhizas geïnfekteer was. Lycorpersicon esculentum cv. FJ44 saailinge was in water kultuur gegroei (pH 5.8) met 0 en 75 mM NaCI asook met of sonder infeksie met die fungus Glomus mosseae. Die plant wortels was bespuit met normale CO2 (360 dele per miljoen (dpm)) sowel as verhoogde CO2 (5 000 dpm) konsentrasies. Die arbuskulere en hife komponente, sowel as die persentasie infeksie was vermeerder of verminder na gelang van die verandering in seisoen. Die plant droë massa van mikorrhiza geïnfekteerde plante by verhoogde CO2 konsentrasies was verhoog met 30% in vergelyking met plante wat nie geïnfekteer was nie, terwyl die droë massa met 68% afgeneem het by gewone CO2 konsentrasies. Verhoogde CO2 konsentrasies het moontlik die plant droë massa en die groei van die fungus verbeter deur koolstof te verskaf as gevolg van die vaslegging van HCO)- deur die werking van PEP karboksilase. Plant wortels wat met verhoogde CO2 konsentrasies bespuit was, het 'n verlaagde CO2 vrystelling getoon in vergelyking met die wortels by normale CO2 vlakke. Die vermindering in CO2 vrystelling van wortels by verhoogde CO2 was die gevolg van die vaslegging van HC03- deur PEPk aktiwiteit. Onder toestande van sout stres, het plante 'n groter hoeveelheid N03- gereduseerde N in die xileemsap bevat in vergelyking met plante wat onder geen sout stres was nie, asook meer NO)- was in die xileemsap vervoer moontlik omdat meer organiese sure geproduseer was ten koste van amino sure. Dit was die moontlike gevolg omdat die plante onder lae P toestande gegroei het. Die tempo van NO.; opname was verhoog onder verhoogde CO2 konsentrasies en in die afwesigheid van sout stres omdat die HCO)- vir die produksie van amino sure gebruik was. In die teenwoordigheid van sout was koolstof moontlik gebruik om organiese sure te vervaardig wat koolstof weggeneem het van die vervaardiging van amino sure. Daar is tot die slotsom gekom dat mikorrhizas voordelig is vir die groei van plante onder toestande van sout stres mits daar 'n addisionele bron van koolstof teenwoordig is. Arbuskulere mikorrhiza infeksie het 'n geringe invloed gehad op die opname van voedingstowwe van plante wat in waterkultuur gegroei was.

There is an urgent need to develop crops that can withstand future climates. Results from this thesis demonstrated that a native Australian resurrection grass exhibits structural, physiological and metabolic strategies to tolerate drying. These strategies may be utilized for the generation of stress tolerant crops.