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Low input grassland systems depend on cheap fertiliser sources, such as clover or animal manures, to maintain sward productivity. Clover is a poor competitor for phosphate (P2O5) and potash (K2O), a mixed sward will therefore require adequate inputs of these nutrients. Nitrogen (N) leads to the suppression of clover. Recycling of cattle slurry would be followed by small N and P2O5 but large K2O effects. The K2O and P2O5 would be expected to have a beneficial effect on the clover while the N would be detrimental. Two investigations into the balance between these aspects of cattle slurry on clover in a mixed sward were performed. Slurry N led to clover suppression, this effect was strongest following spring applications. K2O had a beneficial effect on the clover, the P2O5 supplied had a small positive effect but was insufficient to fully meet the clover's requirements. There was a suggestion that these beneficial effects were strongest following summer applications. In addition to these fertiliser effects, non nutritional secondary effects often followed slurry with a deleterious action on the clover. These effects were most common following higher rates of slurry and summer applications. A further investigation was performed to establish the effect of varying clover variety and companion grass species on the clover's susceptibility to cattle slurry N and secondary effects and the nature of these effects following pig slurry applications. This revealed that clover's susceptibility to slurry N was inversely related to leaf size. No difference in secondary effects susceptibility arose with different clover varieties or companion grass species. Pig slurry was not accompanied by secondary effects. Slurry applications to mixed swards may therefore have positive and negative nutritional effects on the clover component but non nutritional secondary effects may also act with a deleterious effect on the clover.

Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Trace element deficiencies represent an ongoing limitation to agricultural productivity in Southern Australia and in many regions of the world. Trace element deficiencies are commonly encountered on alkaline and calcareous soils due to their high metal adsorption and fixation capacities. Chelating agents, such as EDTA, have been used to reduce fertiliser fixation in these soils and increase trace element transport to the rhizosphere. However, EDTA, which is the most commonly used chelating agent, can be relatively ineffective on alkaline soils and may have negative environmental implications due to its long-term persistence. This study has identified two novel sequestering agents for use on alkaline and calcareous soils. The novel products differ significantly from EDTA in terms of their structure and functionality. For example, rhamnolipid is synthesised by Pseudomonas bacteria, is nontoxic, biodegradable and forms a lipophilic complex with cationic metal ions. The other chelating agent, polyethylenimine (PEl) can complex up to 4 times more metal (g Cu(II)/g ligand) than EDTA, which has important implications for chelate application rates and the cost effectiveness of chelate use. In solution culture experiments, rhamnolipid and PEl facilitated Zn absorption into the root symplast; the kinetic rate of Zn absorption was greater than that of ZnC12 alone. On alkaline and calcareous soils the novel products were significantly (P O.05) more effective Zn sources than EDTA or the SO24-salt. EDTA increased the concentration of Zn in soil solution. However, this did not translate to increased Zn uptake by canola plants. This was not surprising as EDT A inhibited Zn absorption by roots in the solution culture experiments. Radioisotope experiments showed that rhamnolipid and PEl increased Zn adsorption to the soils solid phase. However, PEl increased the size of the total Zn labile pool (P<0.05) and mobilised Zn from the pool of fixed native soil Zn (P<0.05). Rhamnolipid did not significantly (P>O.05) increase the total size of the Zn labile pool in either soil, but significantly (P<0.05) increased Zn uptake by canola, probably by facilitating root absorption by the formation of lipophilic complexes with Zn. These results showed that, on alkaline soils, chelates that increased the rate of trace element absorption into the root symplast were significantly more effective than EDTA, which was not readily absorbed by canola roots. Experiments were also undertaken to explore the effect of chelation on the absorption of foliar applied trace element fertilisers. Perhaps not-unexpectedly, chelation reduced the absorption of foliar applied Zn. The lipophilic chelate, rhamnolipid, quadrupled Zn absorption by enzymatically excised Citrus sinensis cuticles but did not significantly (P>O.05) increase Zn absorption by live leaf tissue. Therefore, there was no discemable relationship between the Kc/w of fertiliser solutions and Zn permeability. This body of work has important implications for future fertiliser development, the cost effectiveness of chelate use and the treatment of micronutrient deficiencies on alkaline soils in the world today. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1272776 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2007

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Biofertilisers are rhizosphere microorganisms inoculated to reduce the need for N or P fertiliser application and maximise plant growth and nutrition, resulting in greater grain yield and N or P content. This study aimed to evaluate the effectiveness of diazotrophic bacteria isolated from the rhizosphere of wheat in Victoria, Australia. This thesis shows that N2-fixing Burkholderia species have great potential as biofertilisers on wheat productivity. In Chapter 2, strains of bacteria were isolated from wheat-growing soils in main Victoria wheat belt at Horsham and Birchip in North West Victoria. Strains were identified as Burkholderia spp. by their closest matches in the 16S DNA and by morphology and physiology. In Chapter 3, one selected strain from each of Birchip and Horsham were used to inoculate wheat in a pot trial in a glasshouse during winter-spring. Soil was collected on site from wheat fields. Pots were inoculated with these strains to evaluate the effects of Burkholderia inoculum as biofertiliser on the plant growth and yield. Different nitrogen sources (urea 46% N and ammonium sulphate 21% N) were used as fertiliser at one of four levels (0, 50, 100 and 150 kg N/ ha). There was a greater effect in Birchip than in Horsham soil and with ammonium sulphate than with urea due to waterlogying in Horsham soil. In Chapter 4, field-grown wheat was inoculated with the same strains of Burkholderia. Three experiments were carried out in plots at two sites, dryland and irrigated fields at Horsham and a dryland field at Birchip, during the winter wheat season of 2006, to evaluate the effect of Burkholderia species inoculum and different types of nitrogen source at one of four levels of added N (0, 50, 100 and 150 kg N/ha) on wheat growth and yield. The effects of both bacterial inoculation and N fertiliser on growth promotion and grain yield. Since 2006 was a year of drought, dry land crops were unsuccessful. Grain %N as well as total N content in grain per area in the Horsham irrigated field increased with increasing N fertiliser levels up to 100 kg N/ha. In Chapter 5, acetylene reduction (ARA) activity was measured in the pots for both inoculated and uninoculated plants at various growth stages and populations of nitrogen-fixing bacteria associated with the wheat roots and bulk soil were measured in addition to biomass and N content of plants and grain. Molecular tracing using specific primers showed that the inoculum was present only in inoculated treatments. Up to 60% of the increased N content of the grain in inoculated plants was potentially derived from nitrogen fixed by the inoculum in the rhizosphere. It was concluded that the most significant result due to inoculation was the consistent maximal increase of N content in grain in inoculated treatments with ammonium sulphate fertiliser at 100 kg N/ha. Inoculation with Burkholderia consistently increased %N in wheat grain, with the potential benefit of decreasing the production cost and reducing use of chemical fertilisers.

The use of urea fertiliser has been associated with relatively poor nitrogen (N) use efficiency (NUE) due to heavy N losses such as gaseous emissions of ammonia (NH₃) and nitrous oxide (N₂O) and nitrate (NO₃⁻) leaching into surface and ground waters. Improving N use-efficiency of applied urea is therefore critical to maximise its uptake and to minimise its footprint on the environment. The study was conducted under laboratory-glasshouse conditions (Chapter 2-4)and lysimiter-field plot studies (Chapter 5). In chapter 2, Two glasshouse-based experimentswere conducted to investigate the potential of incorporating urea fertiliser with ureaseinhibitor, (N-(n-butyl) thiophosphoric triamide (nBTPT) or ‘Agrotain’) to enhance fertiliser N uptake efficiency. Urea, with or without Agrotain, was applied to Ryegrass (Lolium perenne L.) grown in standard plant trays maintained at soil moisture contents of 75–80% field capacity, at rates equivalent to 25 or 50 kg Nha⁻¹. These treatments were compared with other common forms of N fertilisers (ammonium nitrate, ammonium sulphate and sodium nitrate). In a separate pot experiment, granular ¹⁵N urea (10 atom %) with or without Agrotain, was applied at 25 kg Nh⁻¹ to track N use-efficiency and the fate of ¹⁵N-labelled fertiliser. In both experiments, Agrotain-treated urea improved bioavailability (defined as the fraction of total soil N that can interact with a biological target in the plant or that can be taken up by plant) of added N and resulted in significantly higher herbage DM yield and N uptake than urea alone or other forms of N fertilisers. Results from the ¹⁵N experiment support the suggestion that a delay in urea hydrolysis by Agrotain provided an opportunity for direct plant uptake of an increased proportion of the applied urea-N than in the case of urea alone. In chapter 3, two more glasshouse-based experiments were conducted to investigate if urea applied in fine particle application (FPA), with or without Agrotain, had any effect on fertiliser-N uptake efficiency (defined as the difference in N uptake between the fertiliser treatment and the control as a percentage of the amount of N applied) under optimum soil moisture (75-80% field capacity) and temperature (25 °C) conditions, in comparison with other common forms of N fertilisers applied, either in FPA or in granular form. In a separate pot experiment, ¹⁵N urea (10 atom %), with or without Agrotain, was applied to either shoots or leaves only or to the soil surface (avoiding the shoots and leaves) to determine urea hydrolysis, herbage DM and ¹⁵N uptake. In both experiments, herbage DM yield and N uptake were significantly greater in the FPA treatments than in those receiving granular application. Agrotain-treated urea FPA resulted in significantly higher N response efficiency (difference between the dry matter produced by the various fertiliser treatments and the control, divided by the amount of N applied) than urea FPA alone or other forms of N fertilisers. Results from the ¹⁵N experiment support the idea that Agrotain treatment improves the N response of urea applied in FPA form due to a delay in hydrolysis of urea, thus providing herbage an extended opportunity to absorb added urea directly through leaves, cuticles and roots. A further glasshouse-based study was conducted to investigate the effect of Agrotain and irrigation on urea hydrolysis and its movement in a Typic Haplustepts silt loam soil (Chapter 4). A total of 72 repacked soil cores (140 mm inner diameter and 100 mm deep) were used - half (36) of these cores were adjusted to soil moisture contents of 80% field capacity (FC) and the remaining 36 cores to 50% FC. Granular urea, with or without Agrotain, was applied at a rate equivalent to 100 kg N ha⁻¹. Twelve pots were destructively sampled at each day after 1, 2, 3, 4, 7, and 10 days of treatment application to determine urea hydrolysis and its lateral and vertical movement in different soil layers. Agrotain-treated urea delayed urea hydrolysis compared with urea alone during the first 7 days of its application. This delay in urea hydrolysis by Agrotain enabled added urea to disperse and move away from the surface soil layer to the sub-surface soil layer both vertically and laterally. In contrast, most urea in the absence of Agrotain hydrolysed within 2 days of its application. Irrigation after 1 day resulted in further urea movement from the surface soil layer (0-10 mm) to the sub-soil layer (30-50 mm) in Agrotain-treated urea. These results suggest that Agrotain delayed urea hydrolysis and allowed more time for rainfall or irrigation to move the added urea from the surface layer to sub-soil layers where it is likely to make good contact with plant roots. This distribution of urea in the rooting zone (0-200 mm) has the potential to enhance N use efficiency and minimise N losses via ammonia (NH₃) volatilisation from surface-applied urea. Finally, a field study using lysimeters (300 mm inner diameter and 400 mm deep), and small field plots (1 m² in area) was established using a silt loam Typic Haplustepts soil (Soil Survey Staff 1998) to investigate the effect of FPA and granular applications of urea, with or without Agrotain, on N losses and N use efficiency (Chapter 5). The five treatments were: control (no N) and ¹⁵N-labelled urea (10 atom %), with or without Agrotain, applied to lysimeters or mini plots (un-labelled urea), either in granular form to the soil surface or in FPA form (through a spray) at a rate equivalent to 100 kg N ha⁻¹. Gaseous emissions of NH₃ and N₂O, NO₃⁻ leaching, herbage production, N response efficiency, total N uptake and total recovery of applied ¹⁵N in the plant and soil were determined up to 63 days. Urea-alone and urea with Agrotain, applied in FPA form, was more effective than its granular form and reduced N2O emissions by 5-12% and NO3- leaching losses by 31-55%. Urea-alone applied in FPA form had no significant effect in reducing NH₃ losses compared with granular form. However, urea with Agrotain applied in FPA form reduced NH₃ emissions by 69% compared with the equivalent granular treatment. Urea-alone and with Agrotain applied in FPA form increased herbage dry matter production by 27% and 38%, and N response efficiency compared with the equivalent granular urea application, respectively. Urea applied in FPA form resulted in significantly higher ¹⁵N recovery in the shoots compared with granular treatments – this was improved further when urea in FPA form was applied with Agrotain. Thus, treating urea with Agrotain in FPA under field conditions has the potential to delay its hydrolysis, minimise N losses and improve N use efficiency and herbage production. The lower dry matter production and N-response efficiency to urea applied in FPA form in Chapter 3 are probably because of additional factors such as lower application rates (25 kg N ha⁻¹ ) or lack of interception of urea by the leaves. Applying urea in FPA form is a good management strategy and I conclude that combining FPA urea with Agrotain has the potential to increase N use efficiency and herbage production further.

Sheep producers in Victoria are applying more phosphorus (P) fertiliser and increasing stocking rates to increase production. At the same time, there is increasing awareness amongst research, community and producer groups that P-rich runoff water from agricultural land can contribute to the growth of undesirable algal blooms in surface waters. The loads, concentrations and forms of P in surface and subsurface hydrological flow pathways were estimated or measured directly on high and low P fertility hill slope plots in south-west Victoria to assess how this practice change could affect the P status of runoff and drainage water. Small plot rainfall simulator studies were also conducted to investigate pasture management treatment effects. The spatial and temporal distribution of P loss from hillslope pastures, and the processes of P mobilisation in runoff identified in this study were used to identify appropriate management practises to help minimise P losses in runoff. Runoff P concentrations were then related to properties of pasture soils and runoff at a range of locations across Victoria to identify whether simple models and/or soil P tests could be used to predict P concentrations in runoff.