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Janthitrems are believed to be involved in the observed sporadic cases of AR37- infected perennial ryegrass staggers. Investigations into the role of janthitrems in perennial ryegrass staggers are difficult as isolation of the compounds from the ryegrass is hindered by the inherent instability of these compounds. Therefore attempts were made to isolate janthitrems from an alternative source, allowing these janthitrem analogues to be used as surrogates for endophyte produced janthitrems. Analysis of a series of Penicillium janthinellum cultures revealed the presence of janthitrems in a number of strains, including janthitrem B, janthitrem C and two novel janthitrem compounds. Detailed one- and two-dimensional NMR and mass spectral techniques identified the two novel compounds as 11,12- epoxyjanthitrems B and C, which were subsequently given the trivial names janthitrems A and D, respectively. Janthitrems B and C were isolated and identified by NMR and revisions of some previously reported chemical shift assignments were proposed. In addition to the janthitrems, penitrems were also identified in two strains of P. janthinellum. The isolated janthitrem B was utilised for the development of efficient extraction procedures, and for the determination of ideal storage conditions for janthitrem compounds. A method for the extraction and isolation of janthitrem B from a P. janthinellum culture was developed and optimised to yield 6 mg of janthitrem B from 900 mL of fungal culture in two days. Stability studies of janthitrem B indicated the ideal storage condition which minimised degradation was dry at −80 C where only 7% sample loss was observed over 300 days. Bioactivity studies of janthitrems A and B found these compounds to be tremorgenic to mice, with janthitrem A (an epoxyjanthitrem) inducing more severe tremors than janthitrem B. Insect testing also showed that both janthitrems A and B displayed anti-insect activity to porina larvae. Since the epoxyjanthitrems, which are associated with AR37 endophyte-infected ryegrass, were also shown to be tremorgenic and to display anti-insect activity, the insect resistance and the sporadic cases of ryegrass staggers displayed by AR37 may be related to the presence of epoxyjanthitrem compounds. LC-UV-MS analysis of janthitrems A-D, penitrems A-F, lolitrem B, paspalinine, paxilline and terpendole C found these indole-diterpenoids to be more sensitive by analysis using an APCI source as opposed to an ESI source. APCI negative ion LC-UV-MS required source induced dissociation in combination with increased collision energy to suppress an acetate adduct peak, sourced from the acetic acid buffer. Negative ion MS2 and MS3 data produced more informative fragments compared to the conventional positive ion MS2 and MS3 data. The availability of both positive and negative ion LC-UV-MS methodologies will allow future endophyte products to be more thoroughly screened for different classes of secondary metabolites. Extracts of mouldy walnuts were analysed for the presence of tremorgenic mycotoxins after a dog was found to exhibit symptoms characteristic of tremorgenic mycotoxicosis. LC-UV-MS analysis of the mouldy walnuts identified the tremorgenic mycotoxins penitrems A-F, thus confirming the veterinarian's tentative diagnosis of canine tremorgenic mycotoxicosis the first reported case in New Zealand.

A growth room experiment compared seedling growth after nine weeks of two wheat genotypes in pure cultures and mixtures with Italian ryegrass at a low phosphorus (P) level and the P level recommended by soil testing. At the recommended P level in both pure and mixed culture, Mit, a semi-dwarf genotype, had a greater height, leaf area, tiller number, and dry weight of leaves, stems and roots than did Kharkof, a landrace. These results reflected the visual selection criteria for seedling vigor and tillering used in the initial development of the semi-dwarf cultivars. Field experiments compared total dry matter, calories, and percent crude protein and acid detergent fiber of oats, wheat, and ryegrass in pure culture and mixtures at four locations in central Texas at first clipping, second clipping, and unclipped. Although, forage yield was lower for the first clipping than the second clipping or the unclipped treatment, the first clipping had the best forage quality. There was a strong environmental influence on yield and quality. The best forage was pure wheat at College Station, pure ryegrass at Marlin, the wheat-ryegrass mixture at Temple, and pure oats at Thrall. Since Temple had the greatest yield of any site, the wheat-ryegrass mixture demonstrated the highest yield potential. The soil adsorption characteristics of halosulfuron was examined using six soils. Soil adsorption of halosulfuron appeared to be a function of organic matter. Low Kf,ads values indicated that a relatively high plant availability of halosulfuron could be available for plant uptake. Sorghum produced in soils with a low organic matter content may experience injury because of the high plant availability.

Fraze mowing offers a new tool used in both overseeding establishment and removal of perennial ryegrass (Lolium perenne L.). Research objectives were to determine the most effective timing of fraze mowing in overseeding establishment and removal, as well as to compare fraze mowing to other establishment and removal methods. Fall overseeding studies suggest that fraze mowing and scalping in mid-October provides the fastest establishment of ryegrass. However, by mid-March all treatments led to visually similar results. When evaluating removal timings, fraze mowing performed in early- and late-May led to faster visual bermudagrass recovery than March and April timings. Early-season application of glyphosate for ryegrass removal was fairly ineffective; whereas, late-season application was damaging to bermudagrass. Rimsulfuron removed ryegrass more quickly late in the season. Repeated scalping gradually removed ryegrass providing a slower, but complete, bermudagrass recovery. Overall, late-May fraze mowing resulted in the best bermudagrass cover 5 weeks after treatment.

Italian ryegrass is a problematic weed in Mississippi corn production due to the development and proliferation of glyphosate resistance. Studies were conducted to assess Italian ryegrass control prior to planting using herbicides. Effects of fall and spring applied burndown herbicide applications for Italian ryegrass control and subsequent corn grain yield were investigated at the R.R. Foil PSRC in Starkville, MS, at the Coastal Plain Experiment Station in Newton, MS, and the Black Belt Experiment Station in Brooksville, MS on soil textures ranging from sandy loam to silt clay loam. A fall preemergence (PRE) application of S-metolachlor + metribuzin followed by paraquat in the spring provided 99% Italian ryegrass control 28 days after paraquat application. Four spring burndown treatments provided Italian ryegrass control similar to that observed following application of the fall PRE application followed by paraquat in the spring. Applications clethodim + glufosinate + paraquat + dimethenamid-P; clethodim + glufosinate + paraquat + S-metolachlor; clethodim + paraquat + dimethenamid-P; and clethodim + oxyfluorfen + paraquat + S-metolachlor resulted in similar levels of Italian ryegrass control at 96%, 98%, 94%, and 99%, respectively. Corn yield following the fall PRE followed by spring paraquat application was 10,687 kg ha-1. Corn yield following clethodim + paraquat + dimethenamid-P as well as clethodim + oxyfluorfen + paraquat + S-metolachlor applied in the spring resulted in similar corn grain yield to that following the fall PRE followed by spring paraquat application at 9,649 kg ha-1 and 9,567 kg ha-1, respectively. Spring burndown herbicide treatments could be used to control Italian ryegrass while producing similar corn yield to the standard fall herbicide followed by paraquat application in the spring.

Plant form is a compromise between resource gathering, reproduction and the tolerance to physical demands of the abiotic and biotic environment. In an agricultural field in addition to the natural factors causing stress, humans also introduce physical and mechanical stresses, and chemical pesticides into the environment. Many of these factors are hazardous, since they represent stresses to which plants are unable to develop defense mechanisms. However, weeds have persisted in the agricultural environment despite the efforts to eradicate them. They have adapted to environmental changes such as crop rotation and have developed tolerance to stressors like pesticides in very short periods of times (less than 10 years), much less time than normally expected for evolutionary responses to occur. Perhaps a key to why weeds persist in stress-dominated habitats is the way they compromise between yield and survival. The mechanisms that explain which process is relevant in the control of seed production or seedling growth relate to the ecophysiology of the individual plants. However, trade-offs between plants physiological functions will have implications at both population and community levels. Climate change, air pollution and water scarcity are examples of environmental stresses that particularly affect agriculture. Herbicides are a major technological tool for agriculture and are responsible, at least in part, for significant increases in crop production during the last quarter of the century. The research presented in this dissertation was developed to understand the extent that individual responses to multiple environmental stresses can be extrapolated to population-level responses in an annual weed species. The specific objectives were to assess (1) the impact of three anthropogenic stresses (herbicide, UVB light and ozone) and their interactions on individual Italian ryegrass ontogeny and reproduction and (2) the potential evolutionary effect of these stresses and combinations on changes in population size and structure over time. Plants were capable of growth and reproductive compensation under the studied stresses. Stress factors with similar biochemical mechanisms had different effects at the individual plant and population levels of organization. Compensation occurred at all levels of organization: as individuals modifying growth and allocation to different organs and as populations modifying birth, and death rates and density dependent responses. The ability to compensate sometimes decreased with the number of stress factors (e.g. herbicide and UVB). In other cases, compensation ability increased with the number of stress factors (e.g. ozone and herbicide). / Graduation date: 2004

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Phosphorus is a resource in finite supply. Use of organic amendments in agriculture can be a sustainable alternative to inorganic P, provided it can meet crop requirements. However a lack of consistent knowledge of plant P availability following application of organic amendments, limits its potential. Studies suggest chemical extraction procedures, may not reflect plant available P. The Diffusive Gradients in Thin-films (DGT) technique is based on natural diffusion of P via a hydrogel and sorption to a ferrihydrite binding layer; which should accurately represent soil P (CDGT) in a plant available form. The aim of this research was to evaluate changes in soil P availability, following the addition of organic amendments, cattle farmyard manure (FYM), green waste compost (GW), cattle slurry (SLRY) and superphosphate (SP) using Olsen P and DGT. The research included incubation, and glasshouse studies, using ryegrass (Lolium perenne L.). Soils with a history of application of the aforementioned organic amendments were used (Gleadthorpe), as well as a soil deficient in P (Kincraigie). The hypotheses were as follows H1 A build-up of P available by diffusive supply, from historic treatment additions and subsequent availability from fresh treatment additions will be demonstrated by DGT. H2 Historical treatment additions are more important at determining yield and P uptake than fresh additions. H3 DGT can detect changes in P available by diffusive supply following addition of different treatments and subsequently following lysis of microbial cells on a soil deficient in P. H4 DGT will provide a more accurate indication of plant P availability than organic amendments in a soil deficient in P. H5 P measurements using DGT will be lower from organic amendments than superphosphate.H6 DIFS simulations of soil kinetic parameters will provide additional information about how treatments influence P resupply from solid phase to solution following DGT deployment. Cont/d.

In the United Kingdom and Ireland, a major percentage of fertilized agricultural area is devoted to grasslands, which helps to support the associated milk and beef production industries. In temperate grasslands, perennial ryegrass (L. perenne) is the major forage grass and this species is particularly suitable as a forage grass due to its high yield and digestibility, when compared with other species. However, perennial ryegrass is not well adapted to abiotic stress conditions which are likely to occur in its natural environment. Some of the abiotic stress factors which have significant impacts on plant growth and development include water and nutrient availability. Therefore, this project set out to unravel some of the mechanisms involved in the adaptation of perennial ryegrass to limited water, phosphorous and nitrogen. In order to understand the metabolic mechanisms acting in response to these stresses, metabolite profiling was performed using GC-MS. Furthermore, for the water- and phosphorous-limitation studies this approach was complemented with transcript analysis.In order to study water-limitation a hydroponics system supplemented with polyethyleneglycol (PEG) was used to induce water-limitation for a period of one-week. A clear difference in the metabolic profiles of the leaves of plants grown under water stress was observed. Differences were principally due to a reduction in fatty acid levels in the more water stress-susceptible genotype Cashel and an increase in sugars and compatible solutes in the drought-tolerant PI 462336 genotype. Sugars exhibiting a significant increase included, raffinose, trehalose, glucose, fructose and maltose. Raffinose was identified as the metabolite exhibiting the largest accumulation under water-stress in the more tolerant genotype and may represent a target for engineering superior drought tolerance or form the basis of marker-assisted breeding in perennial ryegrass. The metabolomics approach was combined with a transcriptomics approach in the water stress tolerant genotype PI 462336 which identified genes in perennial ryegrass that were regulated by this stress.The characterization of the response to phosphorus-limitation was performed in a hydroponics system containing two solutions with different levels of phosphorus. Samples were collected from the roots and leaves of two genotypes 24 hours after being exposed to stress. Internal phosphate concentrations were reduced and significant alterations were detected in the metabolome and transcriptome of two perennial ryegrass genotypes. Results indicated a replacement of phospholipids with sulfolipids in response to P deficiency and that this occurs at the very early stages of P deficiency in perennial ryegrass. Additionally, the results suggested the role of glycolytic bypasses and the re-allocation of carbohydrates in response to P deficiency The characterization of the metabolic response of L. perenne leaves to different levels of nitrogen supply was performed for seven different genotypes with variability in the regrowth response rate to nitrogen supply in a hydroponics system. This facilitated the identification of common mechanisms of response between genotypes to nitrogen. The metabolic response observed included modifications of the lipid metabolism, as well as alterations of secondary aromatic metabolite precursors in plants exposed to nitrogendeficit. In contrast, plants grown in a nitrogen saturated media appeared to modify to some extent the metabolism of ascorbate. Additionally, it was found that amino acid levels increased with increasing concentrations of nitrogen supplied. This study suggested that the involvement of secondary metabolism, together with lipid and ascorbate metabolism, is of crucial importance in the early-adaptation of perennial ryegrass plants to different levels of nitrogen supply.