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Please read the abstract in the dissertation / Dissertation (MSc (Agric))--University of Pretoria, 2006. / Plant Production and Soil Science / unrestricted

High levels of elements such as sodium (Na), potassium (K) and phosphorus (P) in iron ore minerals are known to reduce the quality and price of these minerals. South Africa, as one of the world largest exporter of iron ore, is affected by this problem. Both potassium (K) and phosphorus (P) are peculiar to South African iron ore. The present study has therefore focussed on developing an environmentally friendly biological method for lowering the levels of K and P in iron ore minerals. Short and long term experiments were set up to isolate, identify, screen and test potential bioleaching bacteria and fungi from different environmental samples. The study started by investigating the possible relationship that exists between weathering and bioleaching processes. The investigation was intended to provide relevant information on the natural role of microorganisms such as ectomycorrhizal (ECM) fungi in the mining environment. The experiments involved the use of both mycorrhizal and non-mycorrhizal Pinus patula seedlings for the weathering of iron ore minerals. Four types of ECM fungi were used, namely Pisolithus tinctorius (PT), Paxillus involutus (PI), Laccaria bicolor (LB) and Suillus tomentosus, (ST). From the results, ectomycorrhizal weathering can be said to be species-specific and significantly influenced by fungal type and particle size. In addition, it was also discovered that both mycorrhizal and non-mycorrhizal roots can participate in weathering processes. Further investigations of ECM fungi when not in symbiosis, were carried out to know how or if they can be potential candidates to mobilise K and P from iron ore minerals. The experimental set up involved in vitro pure cultures of four different ECM fungi, namely Pisolithus tinctorius (PT), Paxillus involutus (PI), Phialocephala fortini (PFR), and Suillus tomentosus (ST). In addition, the treatments involved the use of five different particle sizes of each ore type. The results obtained indicated the potential of the ECM fungi to mobilise P and K from the two iron ore types though at different levels. Factors such as ore type, particle size, organic acid production and attachment of the fungi to the iron ore were all found to influence the mobilisation of nutrients from these ores. Another experiment that addressed some of the limitations encountered with the use of pure cultures of ECM fungi was conducted. Isolated indigenous fungal pure cultures from the surfaces of iron ore minerals were screened for their abilities to solubilise minerals by lowering the levels of K and P. These isolates were identified molecularly as close relatives of three genera that included Penicillium, Alternaria (2 isolates) and Epicoccum for isolates FO, SFC2/KFC1 and SFC2B respectively. The identified Penicillium sp. turned out to be the only phosphate solubiliser among these isolates. Direct bioleaching capability of the fungus was compared to that of its metabolite. At the end, the metabolite showed better K removal than the direct use of the fungi. Interpretation of these results indicates possible relationship between K and P removal, and the organic acids production by this fungus. Other factors such as particle size and mineral type were also found to significantly influence the leaching process. Additional experiment was conducted to investigate the indigeous bacteria and their potentials in reducing the K and P contents of iron ore minerals. A total of 23 bacterial strains that belong to Proteobacteria, Firmicutes, Bacteroidetes and Actinobateria were isolated from the iron ore minerals and identified with molecular methods. All the bacterial isolates were screened for their potential as mineral solubilisers. Only eight of the isolates were selected and used in shake flask experiments that contained both KGT and SK mineral types as their sources of K and P. The experiment showed that all the eight isolates have potentials to produce organic acids especially high levels of gluconic acid but lower quantities of acetic, citric and propanoic acid. Scanning electron microscopy (SEM) and fourier transform infrared (FITR) analyses also helped to uncover the role that biofilm and extracellular polymeric substances could play in mineral solubilisation. Finally, an investigation of a new method for reduction of K and P levels of iron ore minerals was carried out, focussing on the use of cheap resources as well as septic conditions. The study involved the use of fermented spoilt grape fruits (Vitis sp.) and the solution from the product utilised in shake-flask experiments. Treatments involved two types of iron ore minerals (KGT and SK) and two different particle sizes. The result suggests the significant effect of particle size, time and organic acids on the reduction of K and P from the iron ore minerals. The important part of this finding is the discovery of a cheap microbial energy source (spoilt grape) that can be further exploited for full biobeneficiation of iron ore minerals. Another advantage of this method is the fact that the experiment can be conducted under non–sterile conditions, making it a system that can be operated outdoor. / Thesis (PhD)--University of Pretoria, 2010. / Microbiology and Plant Pathology / unrestricted

The rapid accumulation of municipal solid waste is a significant environmental concern in our rapidly growing world. Due to its low cost, high energy recovery and limited environmental impact anaerobic digestion (AD) is a promising solution for stabilizing the organic fraction of municipal solid waste (OFMSW). Hydrolysis is often the rate-limiting step during AD of wastes with high solid content; this step can be accelerated by pretreatment of waste prior to AD. This thesis presents the results of alkaline pretreatment of OFMSW using NaOH and KOH. Four different pH levels 10, 11, 12 and 13 at two temperatures 23±1°C and 80±1°C were examined to study the effects of the pretreatment on (i) enhancing the solubility of the organic fraction of the waste, and (ii) enhancing the AD process and the biogas production. The effects on solubility were investigated by measuring changes in the soluble COD (SCOD) concentrations of pretreated wastes and the enhanced AD was investigated by measuring volatile solids (VS) destruction, total COD (TCOD) and SCOD removal in addition to biogas and methane production using biochemical methane potential (BMP) assay and semi-continuous laboratory reactor experiments. Pretreatment at pH 13 at 80±1°C demonstrated the maximum solubility for both NaOH and KOH pretreated samples; however the BMP analysis demonstrated that pretreatment at pH 12 at 23±1°C showed the greatest biogas yield relative to the removed VS for both chemicals. Thus pretreatment at pH 12 at 23±1°C using NaOH and KOH were examined using semi-continuous reactors at three different HRTs: 10, 15 and 20 days. Pretreatment demonstrated a significant improvement in the AD performance at SRTs of 10 and 15 days.

Experiments were undertaken to examine the extent of variation for potassium and nitrogen uptake and utilization in wheat and also to address some issues of relevance to the improvement of these traits. These issues included the inheritance of these traits and the difficulties that could arise due to (1) the methodology that is used to measure ion fluxes and utilization, (2) ontogenetic variation in the expression of these traits, and (3) the growth stage at which nutrient utilization is evaluated. To compare varieties developed during different periods in the history of wheat breeding, the varieties were assigned to five groups on the basis of height and origin. Nutrient fluxes were measured either as average net fluxes or short-term net fluxes. Nutrient utilization was expressed as shoot fresh weight per plant, efficiency ratio or utilization efficiency. Substantial variation was observed for all traits except potassium and nitrogen efficiency ratios. Although short-term net potassium fluxes were negatively correlated with root potassium concentration, some of the differences in flux were not associated with differences in root potassium concentration. These differences must therefore be heritable. Due to the complexity of the regulation of nitrate uptake, genotypic differences in short-term net nitrate flux were not examined in relation to root nitrate concentration. Therefore, some of the variation in nitrate flux could be due to differences in root nitrate concentration or some other factor(s) which regulates nitrate uptake. Significant differences between groups were also observed. The tall varieties had the highest potassium and nitrate fluxes but were not significantly different from the triple dwarfs. The double dwarfs were the poorest performers for both nutrient uptake and utilization. In general, the tall traditional varieties were more vigorous and hence showed the highest shoot weight per plant and utilization efficiencies. These findings are examined in relation to the contention that plant breeding under high fertility conditions may have resulted in a decline in the ability of plants to acquire and utilize mineral nutrients. The inheritance of short-term net potassium flux, shoot weight per plant, potassium efficiency ratio and potassium utilization efficiency was studied in four crosses. Complex modes of inheritance were observed for all the traits. For one of the crosses significant reciprocal effects were observed for shoot weight per plant, efficiency ratio and utilization efficiency. Narrow sense heritabilities for the two traits most likely to be selected for, namely short-term net potassium flux and shoot weight per plant, indicated that selection for these traits should be carried out amongst families rather than amongst single plants. Diallel analysis for nitrate uptake and utilization indicated that both additive and dominance gene effects are important in the determination of these traits. The effect of developmental changes in potassium uptake and utilization on varietal comparisons and genetic studies was investigated by comparing the performance of six varieties at different stages of growth over a five-week period. The rankings of the varieties for short-term net potassium flux and shoot weight per plant were found to be fairly consistent. Correlations between average net fluxes for different time periods as well between short-term and average net fluxes were poor. These findings indicate that selection for differences in uptake should be based on fluxes obtained from solutions identical in concentration to the growth solution rather than on perturbation fluxes obtained by depletion of a solution much more concentrated than the growth solution. All measures of potassium utilization based on vegetative growth were poorly correlated with performance at the adult stage. Significant negative rank correlations between shoot fresh weight per plant and grain weight per plant were obtained most likely due to differences in harvest index. This finding casts some doubt on the usefulness of vegetative measures of nutrient utilization as indicators of nutrient-use efficiency for a crop in which the economic product consists of grain. / Science, Faculty of / Botany, Department of / Graduate

Orientador: Rogério Peres Soratto / Coorientador: Adalton Mazetti Fernandes / Banca: Thiago Leandro Factor / Banca: Roberto Lyra Villas Boas / Resumo: Com o uso de cultivares de batata (Solanum tuberosum L.) mais produtivas, possivelmente a demanda por nutrientes também ficou maior, levando a necessidade de se adequar a adubação. Dentre os nutrientes absorvidos pela cultura, destaca-se o potássio (K), que é extraído em grande quantidade, e é de extrema importância para o desenvolvimento da cultura, para a elevação da produtividade e para proporcionar tubérculos de maior qualidade. Muitos produtores estão adotando adubações sem recomendação técnicas e, como o K é muito extraído pela cultura, as baixas doses de K e manejo inadequado podem estar limitando a produtividade, principalmente em áreas com baixos teores de K disponível. Diante do exposto, o objetivo deste trabalho foi avaliar o efeito de doses e do parcelamento da adubação potássica na nutrição e produtividade da cultura da batata, cv. Ágata. Foram realizados três experimentos em campo, em solos de textura argilosa e com teores baixo (0,7 mmolc dm-3), médio (1,6 mmolc dm-3) e alto (3,7 mmolc dm-3) de K trocável. Os experimentos foram conduzidos no delineamento experimental de blocos casualizados, em esquema fatorial 3x2+1, com quatro repetições. Os tratamentos foram compostos pela combinação de três doses de K (100, 200 e 400 kg ha-1 de K2O), com duas formas de parcelamento (100% no sulco de plantio e 50% no sulco + 50% em cobertura, por ocasião da amontoa) e mais uma testemunha (sem aplicação de K). Independentemente do parcelamento, o aumento das doses de K proporcionou incremento no teor de K na folha da batateira, cultivada em solos com baixa, média e alta disponibilidade de K, porém, de forma mais acentuada no solo com baixo teor. No solo com baixo teor de K trocável, a aplicação da dose K totalmente no sulco de plantio proporcionou maior teor de K na folha do que quando parcelado no plantio e em cobertura. Independentemente da disponibilidade de ... / Abstract: With the use of more productive potato (Solanum tuberosum L.) cultivars, the plant's demand for nutrients has possibly increased, raising the necessity of adjustments on fertilizer application. Potassium (K), which is extracted in high amounts, is one of the main nutrients absorbed by the plant. This compound is of pivotal importance to potato development, especially to increase its yield and to generate high quality tubers. Many producers are making use of fertilization without a proper technical recommendation. As K uptake by the plant is very high, low K rates and inadequate management can impair tuber yield, mainly in soil with low K availability. In this context, this study aimed to evaluate the effects of K fertilization rates and split application on nutrition and yield or potato crop, cv. Agata. For this, three field experiments was carried out on clay-textured soils with low (0.7 mmolc dm-3), medium (1.6 mmolc dm-3) and high (3.7 mmolc dm-3) exchangeable K concentration. These experiments was conducted under a randomized block design using a 3x2+1 factorial scheme, with four replications. The treatments consisted of three K rates (100, 200, and 400 kg ha-1 K2O), combined with two forms of split application (100 % at planting furrow or 50% at planting furrow + 50% at sidedressing, during hilling) and a control (without K application). Regardless of installments, increasing K rates showed an increase in K concentration in leaf of potato grown in soils with low, medium, and high K availability, but more markedly in soil with low availability. In soil with low concentration of exchangeable K, the application of K rate totally at planting furrow increased the concentration of K in the leaf than when split applied at planting and hilling. Regardless of the soil K availability, K fertilization reduced the concentrations of P, Ca, and Mg in leaf of potato crop and little influenced the ... / Mestre

Adenylyl Cyclases (ACs) catalyze the formation of the key universal second messenger adenosine 3’, 5’-cyclic monophosphate (cAMP) from adenosine 5’- triphosphate. Cyclic AMP participates in several signal transduction pathways and is present in bacteria and higher and lower eukaryotes including higher plants. Previous studies in plants have shown a role for cAMP in signal transduction during e.g. the cell cycle, elongation of the pollen tube and stimulation of protein kinase activity. More recently cAMP has been shown to play a role in stress responses. Interestingly, cAMP has also been shown to regulate ion transport in plant cells. Here we used a similar strategy that led to the discovery of the first guanylyl cyclase in plants that was based on the alignment of conserved and functionally assigned amino acids in the catalytic centre of annotated nucleotide cyclases from lower and higher eukaryotes, to identify a novel candidate ACs in Arabidopsis (Arabidopsis thaliana K+ Uptake 5 and 7). ATKUP5 and 7 are homologous to K+ uptake permeases (KUPs) from bacteria and high-affinity K+ transporters (HAKs) from fungi. The AC activity was investigated by recombinantly expressing the ATKUP5 and 7 AC domain in vitro and by complementation of an E. coli AC mutant (cyaA). Furthermore, ATKUP5 was tested for its ability to functionally complement a yeast mutant deficient in Trk1 and Trk2 high affinity potassium uptake transporters. Site-mutagenesis in the AC domain was used to test the effect of both functions in each other. Furthermore, ATKUP5 was characterized electrophysiologically in HEK-293 cells to characterize the nature of this transporter. The localization of the ATKUP5 in Arabidopsis was examined using a Green Fluorescent Protein (GFP) fusion with the ATKUP5 to determine whether ATKUP5 is expressed at the plasma or tonoplast membrane. Arabiodpsis thaliana of the wild type, overexpressing ATKUP5 and atkup5 mutant lines were used to examine phenotypic differences.

Tartrate stabilization is the process that removes components that contribute to the crystallization of potassium hydrogen tartrate (KHT) and calcium tartrate (CaT) which is an undesirable outcome for wine quality. There are a variety of current tartrate stabilization techniques such as cold stabilization, chemical additives, ion exchange resins, and electrodialysis that stabilize wine, but the most popular being cold stabilization. Cold stabilization requires high amounts of energy and resources to stabilize wine. With the ever increasing demand for more efficient processing, an alternative tartrate stabilization technology based on an electrolytic method was developed and its viability to stabilize wine was determined. Twelve treatments involving different combinations of time and current were replicated three times each on different batches of Chardonnay wine. Several different variables were analyzed for stability and quality purposes. Tartaric acid, potassium, calcium, and conductivity differences were the most important factors for tartrate stability. Temperature, titratable acidity, pH, color (hue and intensity), and chemical oxygen demand (COD) were indicators of sensory quality characteristics of the wine. The concentrations of potassium, calcium, and tartaric acid were reduced by the electrolytic method at satisfactory process parameters, inherently making the wine more stable. The temperature and hue were significantly affected by the electrolytic method and accelerated the oxidative browning process. Electrolytic treatment of Chardonnay is a viable alternative stabilization technology. The technology can be further developed to become a great option in terms of water and energy consumption, process time, and price.

Aim. The aim of the study was to evaluate the efficacy, safety and possible mechanism of action of potassium humate loaded with selenium Objectives. The objectives of the study were to evaluate the possible in vitro cytotoxic effect of potassium humate loaded with selenium (Phse) on the growth of primary cell cultures (lymphocytes), to evaluate the in vitro antioxidant activity of Phse, to evaluate the in vitro effect of Phse on CR3 expression using mixed leukocytes, to evaluate the anti-inflammatory properties of Phse using the carrageenan-induced paw oedema rat model and finally to evaluate the effect of Phse on acute phase proteins in the rat model Methods. For the cytotoxicity effects on lymphocytes, the MTT assay was used where lymphocytes were isolated and divided in to two groups, one group was stimulated with PHA and the other not, then the cells were treated with different concentrations of the test compounds. The evaluation of the antioxidant activity was done using the ORAC and DCFH-DA assays with the DCFH-DA assay also done using the HepG2 cell line. The expression of CR3 by mixed leukocytes was quantified by flow cytometry. The evaluation of the anti-inflammatory properties of Phse was done using the carrageenan-induced paw oedema rat model. The rats were randomly assigned to six groups, the negative control, positive control, experimental group 1, 2, 3 and sham group. A once daily dose by gavage for five consecutive days of their respective treatment was administered. Prior to the assay the rats were dosed according to the experimental group to which they were assigned. On the fifth day of the experiment, 50 µl of λ- Carrageenan was injected subplanter into the right hind paw of the rats and 50 µl of saline into the left hindpaw. The right hind paw volume of each rat was measured hourly from the time of injection for seven consecutive hours with a water displacement plethysmometer. At the end of the 7 hours the rats were anaesthetised and approximately 5 ml of blood was collected via cardiac puncture. The blood was centrifuged, the plasma removed and frozen at -80°C until assayed. For evaluating the effects of the test compounds on acute phase proteins ELISA was used according to the manufacturer’s protocols. Results. None of the test compounds were toxic to lymphocytes but rather caused cell proliferation. The test compounds demonstrated no antioxidant activity, with Phse showing pro-oxidant activity. All the test compounds inhibited the expression of CR3 significantly with selenium free Ph being the most potent inhibitor. Ph reduced the carrageenan induced paw oedema volumes in a similar manner to indomethacin and Phse had an insignificant effect. Ph decreased SP slightly but the results were not statistically significant whereas Se and Phse had no effect. All the test compounds statistically significantly decreased plasma CRP levels with Se showing the greatest effect. Conclusion. Phse is safe but not more effective than Ph as an anti-inflammatory agent. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Pharmacology / unrestricted