Twenty-nine Rhizobium leguminosarum biovar trifolii strains were tested for acidity tolerance in acidified liquid medium. Only 41\ of the strains grew at pH 4.1. One acid-tolerant strain, USDA 2160, and one acid-sensitive strain, 162-X-103 from Nitragin Co., were inoculated on seeds of 'Nungarin', 'Seaton Park' and 'Clare' subclover cultivars. The inoculated and pelleted seeds were sown in potted Cluff soil with pH 5. 7. Three desiccation levels were imposed by delaying watering for 0, 15 or 30 days. Four gravimetric soil water contents (6.0, 6.6, 10.5 and 12.5\) were maintained under a greenhouse line-source sprinkler system for 7 weeks. The desiccation treatments were more detrimental to the survival of the acid-tolerant Rhizobium strain (USDA 2160) than they were for the acid sensitive strain (162-X-103). Symbiotic effectiveness, measured as shoot dry weight, was higher with strain 162-X- 103 than with strain USDA 2160 and was comparable to the Nfertilized control at the highest water level (12.5 %). At the lower water levels (6.0, 6.6 %) symbiotic N2-fixation was more affected than N-uptake. The Rhizobium strains were able to survive and grow even at the lowest soil water level. The number of rhizobia in the soil and nodulation of the subclover plants had a correlation of 0.56.

No description available.

Lolium perenne cv. Nui is a cultivar of ryegrass grown throughout New Zealand in pastures due to favourable traits such as high palatability for livestock and its ability to withstand intensive grazing. However, the productivity of pastures is reduced when levels of aluminium and other metals accumulate in soils to toxic levels, a phenomenon referred to as the ‘acid soil syndrome’. In response to this toxicity, plants activate a series of antioxidant reactions, with one catalysed by the superoxide dismutase (SOD) enzymatic system. The enzyme system comprises three isoenzymes, a Cu/ZnSOD, FeSOD and a MnSOD which catalyse the same reaction but differ in amino acid sequence, molecular mass and the metal ion co-factor (hence Cu/ZnSOD, FeSOD and MnSOD). Together these isoenzymes combat the damaging effect of superoxide radicals which accumulate due to metal toxicity. In this thesis, the isolation of genes encoding isoenzymes of the SOD enzyme from L. perenne cv. Nui is described. As well, the growth of L. perenne cv. Nui and changes in expression of the SOD genes encoding each isoenzyme in response to aluminium treatment (0.2mM AlCl3) is investigated. A 1072 bp FeSOD gene sequence and a 705 bp MnSOD gene sequence were isolated from shoot tissue of L. perenne cv. Nui using a combination of RT-PCR with degenerate primers and 3'-RACE. The FeSOD gene comprised 572 bp of the coding sequence and 500 bp of 3'-UTR while the MnSOD gene comprised 508 bp of coding sequence and a 197 bp 3'-UTR. By alignment of each sequence with the gene from the database with highest identity it was predicted that the translation start codon (ATG) is located a further 196 bp upstream for the FeSOD gene (aligned with an Oryza sativa FeSOD sequence as a reference) and a further 152 bp upstream for the MnSOD sequence (aligned with a Triticum aestivum MnSOD sequence as a reference). Using RT-PCR with degenerate primers, a 313 bp CuSOD sequence was predominantly cloned from shoot tissue of L. perenne cv. Nui, but it was not possible to generate the 3'-UTR using 3'-RACE. For growth analysis, seedlings of L. perenne cv. Nui were germinated and acclimatised in Hoagland’s solution, and then subjected to either aluminium treatment (0.2mM AlCl3) or no treatment to act as a control over a designated time course of 0, 4, 8, or 24 hours. Two growth trials were conducted that differed in the age of seedlings used and plant tissues were separated into root and shoot tissues. Similar growth trends were observed in both trials, but the sampling regime in the second growth trial meant that statistical analysis could be carried out. In this trial, analysis revealed that over a time course of 24 hours exposure to 0.2mM aluminium, both root and shoot tissue fresh weight did not significantly differ when compared to the control (no aluminium). A general trend of an increase in root and shoot fresh weight was observed in plants treated with aluminium, but this trend was not significant at P=0.05. No significant change in fresh weight partitioning from shoot to root, or root to shoot in response to aluminium was also observed. Using semi-quantitative Reverse Transcriptase-Polymerase Chain Reaction (sqRTPCR) and primers based around the 3'-UTR with RNA isolated from plants grown in the second hydroponic trial, it was determined that under the conditions used, expression of the FeSOD and MnSOD genes isolated in this study were neither up-regulated or downregulated in response to aluminium treatment in both shoot and root tissue. Further, using degenerate primers to detect expression of one or more genes encoding the Cu/ZnSOD isoenzyme, total expression of the Cu/ZnSOD isoenzyme was also unresponsive to aluminium treatment.

Lolium perenne cv. Nui is a cultivar of ryegrass grown throughout New Zealand in pastures due to favourable traits such as high palatability for livestock and its ability to withstand intensive grazing. However, the productivity of pastures is reduced when levels of aluminium and other metals accumulate in soils to toxic levels, a phenomenon referred to as the ‘acid soil syndrome’. In response to this toxicity, plants activate a series of antioxidant reactions, with one catalysed by the superoxide dismutase (SOD) enzymatic system. The enzyme system comprises three isoenzymes, a Cu/ZnSOD, FeSOD and a MnSOD which catalyse the same reaction but differ in amino acid sequence, molecular mass and the metal ion co-factor (hence Cu/ZnSOD, FeSOD and MnSOD). Together these isoenzymes combat the damaging effect of superoxide radicals which accumulate due to metal toxicity. In this thesis, the isolation of genes encoding isoenzymes of the SOD enzyme from L. perenne cv. Nui is described. As well, the growth of L. perenne cv. Nui and changes in expression of the SOD genes encoding each isoenzyme in response to aluminium treatment (0.2mM AlCl3) is investigated. A 1072 bp FeSOD gene sequence and a 705 bp MnSOD gene sequence were isolated from shoot tissue of L. perenne cv. Nui using a combination of RT-PCR with degenerate primers and 3'-RACE. The FeSOD gene comprised 572 bp of the coding sequence and 500 bp of 3'-UTR while the MnSOD gene comprised 508 bp of coding sequence and a 197 bp 3'-UTR. By alignment of each sequence with the gene from the database with highest identity it was predicted that the translation start codon (ATG) is located a further 196 bp upstream for the FeSOD gene (aligned with an Oryza sativa FeSOD sequence as a reference) and a further 152 bp upstream for the MnSOD sequence (aligned with a Triticum aestivum MnSOD sequence as a reference). Using RT-PCR with degenerate primers, a 313 bp CuSOD sequence was predominantly cloned from shoot tissue of L. perenne cv. Nui, but it was not possible to generate the 3'-UTR using 3'-RACE. For growth analysis, seedlings of L. perenne cv. Nui were germinated and acclimatised in Hoagland’s solution, and then subjected to either aluminium treatment (0.2mM AlCl3) or no treatment to act as a control over a designated time course of 0, 4, 8, or 24 hours. Two growth trials were conducted that differed in the age of seedlings used and plant tissues were separated into root and shoot tissues. Similar growth trends were observed in both trials, but the sampling regime in the second growth trial meant that statistical analysis could be carried out. In this trial, analysis revealed that over a time course of 24 hours exposure to 0.2mM aluminium, both root and shoot tissue fresh weight did not significantly differ when compared to the control (no aluminium). A general trend of an increase in root and shoot fresh weight was observed in plants treated with aluminium, but this trend was not significant at P=0.05. No significant change in fresh weight partitioning from shoot to root, or root to shoot in response to aluminium was also observed. Using semi-quantitative Reverse Transcriptase-Polymerase Chain Reaction (sqRTPCR) and primers based around the 3'-UTR with RNA isolated from plants grown in the second hydroponic trial, it was determined that under the conditions used, expression of the FeSOD and MnSOD genes isolated in this study were neither up-regulated or downregulated in response to aluminium treatment in both shoot and root tissue. Further, using degenerate primers to detect expression of one or more genes encoding the Cu/ZnSOD isoenzyme, total expression of the Cu/ZnSOD isoenzyme was also unresponsive to aluminium treatment.

Lolium perenne cv. Nui is a cultivar of ryegrass grown throughout New Zealand in pastures due to favourable traits such as high palatability for livestock and its ability to withstand intensive grazing. However, the productivity of pastures is reduced when levels of aluminium and other metals accumulate in soils to toxic levels, a phenomenon referred to as the ‘acid soil syndrome’. In response to this toxicity, plants activate a series of antioxidant reactions, with one catalysed by the superoxide dismutase (SOD) enzymatic system. The enzyme system comprises three isoenzymes, a Cu/ZnSOD, FeSOD and a MnSOD which catalyse the same reaction but differ in amino acid sequence, molecular mass and the metal ion co-factor (hence Cu/ZnSOD, FeSOD and MnSOD). Together these isoenzymes combat the damaging effect of superoxide radicals which accumulate due to metal toxicity. In this thesis, the isolation of genes encoding isoenzymes of the SOD enzyme from L. perenne cv. Nui is described. As well, the growth of L. perenne cv. Nui and changes in expression of the SOD genes encoding each isoenzyme in response to aluminium treatment (0.2mM AlCl3) is investigated. A 1072 bp FeSOD gene sequence and a 705 bp MnSOD gene sequence were isolated from shoot tissue of L. perenne cv. Nui using a combination of RT-PCR with degenerate primers and 3'-RACE. The FeSOD gene comprised 572 bp of the coding sequence and 500 bp of 3'-UTR while the MnSOD gene comprised 508 bp of coding sequence and a 197 bp 3'-UTR. By alignment of each sequence with the gene from the database with highest identity it was predicted that the translation start codon (ATG) is located a further 196 bp upstream for the FeSOD gene (aligned with an Oryza sativa FeSOD sequence as a reference) and a further 152 bp upstream for the MnSOD sequence (aligned with a Triticum aestivum MnSOD sequence as a reference). Using RT-PCR with degenerate primers, a 313 bp CuSOD sequence was predominantly cloned from shoot tissue of L. perenne cv. Nui, but it was not possible to generate the 3'-UTR using 3'-RACE. For growth analysis, seedlings of L. perenne cv. Nui were germinated and acclimatised in Hoagland’s solution, and then subjected to either aluminium treatment (0.2mM AlCl3) or no treatment to act as a control over a designated time course of 0, 4, 8, or 24 hours. Two growth trials were conducted that differed in the age of seedlings used and plant tissues were separated into root and shoot tissues. Similar growth trends were observed in both trials, but the sampling regime in the second growth trial meant that statistical analysis could be carried out. In this trial, analysis revealed that over a time course of 24 hours exposure to 0.2mM aluminium, both root and shoot tissue fresh weight did not significantly differ when compared to the control (no aluminium). A general trend of an increase in root and shoot fresh weight was observed in plants treated with aluminium, but this trend was not significant at P=0.05. No significant change in fresh weight partitioning from shoot to root, or root to shoot in response to aluminium was also observed. Using semi-quantitative Reverse Transcriptase-Polymerase Chain Reaction (sqRTPCR) and primers based around the 3'-UTR with RNA isolated from plants grown in the second hydroponic trial, it was determined that under the conditions used, expression of the FeSOD and MnSOD genes isolated in this study were neither up-regulated or downregulated in response to aluminium treatment in both shoot and root tissue. Further, using degenerate primers to detect expression of one or more genes encoding the Cu/ZnSOD isoenzyme, total expression of the Cu/ZnSOD isoenzyme was also unresponsive to aluminium treatment.

Thesis (MSc. Agriculture(Soil Science) -- University of Limpopo, 2018 / Soil acidity is one of the abiotic stress factors that greatly limit the productivity of crops on farmers’ fields. A greenhouse study was carried out over two summer growing seasons to evaluate the effect of lime and phosphorus (P) application rates on the growth, yield and root attributes of a determinate cowpea variety on acid soil. The experiment was laid out as a 4x5 factorial arrangement with 4 replications. Treatment factors comprised of variable rates of Vaalburg dolomitic lime (0, 2, 4 and 6 t ha-1) and P (0, 15, 30, 45 and 60 kg ha-1) using single super phosphate, 10.5% P. The two treatment factors were combined resulting in a total of 20 treatment combinations. Data collected included cowpea growth parameters, crop phenology, yield attributes and root characteristics. While cowpea plants with no P application consistently gave the least plant height, stem diameter, number and length of trifoliate leaves, the 6 t ha1 lime rate appears to be completely disadvantaged for all measured parameters with generally lower values than in soil filled pots without lime application. Results showed that soil pH was increased with 6 t ha-1 lime application while soil electrical conductivity (EC), percent of organic matter (OM) and total organic carbon (TOC) were all increased with increasing P and lime rates. All measured cowpea growth attributes such as plant height, stem diameter, number of trifoliate leaves, and leaf area were significantly increased (p≤0.05) with increasing P and lime rates. During the two planting seasons, P and lime application resulted in reduced (p≤0.05) duration to flowering, pod formation and physiological maturity. The 6 t ha-1 lime application produced higher number of pods (2.50) compared to the other rates. Application rates of 45 kg P ha-1 and 6 t ha-1 of lime produced superior number of seeds per pod with high values of (13.71) and (12.85), respectively. However, cowpea root attributes namely number of nodules per plant, the third branching root diameter, angle of adventitious root, tap root diameter at 5 and 10 cm, shallow and deep score were significantly increased at moderate P rate of 30 kg P ha-1. Overall, findings of this study revealed that application of both P fertiliser and lime were able to ameliorate the negative effect of P deficiency from soil acidity on the evaluated cowpea variety and promoted increased yield. Keywords: Acid soil, grain cowpea, P fertiliser, lime, growth, root characteristics, yield / National Research Foundation (NRF) and Department of Agriculture, Forestry and Fisheries (DAFF)

Nitrogen is one of the major elements essential for plant growth. It is the element most prone to depletion by cultivation and is usually found deficient in soils. In successful farming the problem of nitrogen supply is of special importance because plants need it in large amounts, it is fairly expensive to supply, and it is easily lost from the soil.

No description available.

Doctor of Philosophy / Department of Agronomy / Ganga M. Hettiarachchi / Although phosphorus (P) is an essential nutrient for the growth of plants, it is one of the most limiting nutrients in terms of availability as a high proportion of applied P rapidly transforms into insoluble forms with low solubility in soils. To further understand the fate of P applied to soils, two separate but related studies using three high P-fixing soil types each were used for which the objectives were to investigate the mobility, availability, and reaction products from two granular and one liquid P fertilizer alone or plus a fertilizer enhancement product. Energy dispersive spectroscopy showed a substantial amount of P remained in the granule following a 5-week incubation. At the end of the 35-day incubation period there was evidence that the fluid fertilizer was superior over the granular sources in terms of enhanced diffusion and extractability of P for three calcareous soils with varying levels of CaCO₃. Phosphorus x-ray absorption near-edge structure (XANES) spectroscopy results in conjunction with resin-extractable P indicated a strong negative correlation between Ca-P solids formed and P extractability, suggesting that degree of Ca-P formation limits P solubility. For the three acidic P-fixing soils the results were complex. In two out of three acid soils, liquid P treatments diffused farther from the application point than the granular treatments. Phosphorus XANES results suggested that Fe-P or Al-P interactions control the overall P solubility. Integration of pH, resin extractable-P and XANES results suggested the P retention mechanism was either dominated by adsorption or precipitation depending on soil pH. More acidic soil conditions favored precipitation. The objectives of the third study were to observe how long-term (14 years) addition of P with or without N influences the inorganic and organic P pools in a native grassland soil using sequential fractionation, XANES, and ³¹P-nuclear magnetic resonance (NMR) spectroscopy. The overall results suggested that P and N fertilization and associated changes in plant productivity induced significant changes in soil P pools such as Ca-P, phytic acid, monoesters, and residual forms of P. The addition of P alone induced formation of inorganic P forms while the addition of P and N induced transformation of residual P forms into more labile and/or organic P forms.

[Truncated abstract] Standard AOAC methods of chemical analysis have been used to characterize and evaluate the industrial byproducts; partly burnt chicken litter ash (CLA), totally burnt chicken litter ash (CLAT), wood ash (WA) and iron smelting slag for use as a combined liming agent and phosphate fertilizer. Rock phosphate has this function and was included for comparison purposes. All the byproducts had pH values above 9 and a liming capacity above 90% of pure lime, as a result, these materials will be effective as liming agents. Total P concentrations for CLA, CLAT, slag, and WA were 3.6%, 4.75%, 0.26%, and 0.44% respectively indicating that they could be used as P fertilizers when applied at the high rates required for liming soils. ... The RE values for all the materials relative to monocalcium phosphate (100%) for the first harvest are as follows, 50% for dicalcium phosphate, 31% for rock phosphate, 7% for partly burnt chicken litter ash, 7% for totally burnt chicken litter ash and 1% for wood ash and slag. The RE values for the second harvest were 100% for monocalcium phosphate, 80% for dicalcium phosphate, 40% for rock phosphate, 10% for partly burnt chicken litter ash, 8% for totally burnt chicken litter ash and 2% for wood ash and slag. Data for subsequent harvests are not reported due to the death of many plants. Clearly chicken litter ash has appreciable value as a phosphate fertilizer whereas wood ash and slag are ineffective. Explanations for these differences in effectiveness are discussed in the text. An evaluation of the liming effect of the byproducts indicates that they may be used as a soil amendment on acid soils and are nearly as effective as standard lime (CaCO3). Byproducts are also sources of other plant nutrients so they may be regarded as a form of compound fertilizer and liming agent.