Penetration of sink as well as source leaves of wheat plants by the Russian wheat aphid, Diuraphis noxia (Mordvilko) was investigated using light, fluorescence and transmission electron techniques, to determine the feeding strategies adopted by the aphid in penetrating and successfully feeding from the phloem, and to assess the structural effects of the probing and feeding behaviour of D. noxia on the feeding sites. Examination of aphid-infested sink, as well as source leaf tissue, showed that D.noxia probed in cells of the vascular bundle more frequently than mesophyll cells. Within the vascular bundle, thin-walled sieve tubes were visited (probed) more than the other cells. In sink leaf material, 68 of 82 (83%) stylets and stylet tracks encountered during the examination of 1000 serial sections (from 5 different plants) terminated in thin-walled sieve tubes and only 14 (17%) in thick-walled sieve tubes. Thin-walled sieve tubes were visited more significantly than thick-walled sieve tubes. However, examination of the aphid.,.infested sink leaf on a per centimetre basis, from the tip of the leaf, revealed that thick-walled sieve tubes in the area closest to the tip (0-2cm from the tip) were as attractive to the aphid as were thin-walled sieve tubes, with no significant difference in the number of times thick- and thin-walled sieve tubes were probed in this area. Some 2-4cm from the tip however, thinwalled sieve tubes were significantly more probed and therefore more attractive than thick-walled sieve tubes. Examination of 2000 serial sections using aphid-infested source leaf tissue, showed that the thin-walled sieve tubes were significantly more probed than thickwalled sieve tubes, along the whole leaf, expressed as a total of all leaves, as well as on a per centimetre basis along the length of the leaf, with 212 (95%) of 222 terminations within the thin-walled sieve tubes and only 10 (5%) in thick-walledsieve tubes. The aphid probed the small vascular bundles (loading bundles) many more times than intermediate or large transport vascular bundles, in sink as well as source leaf. Of a total of 82 stylets and stylet tracks encountered in sink leaf tissue, 31 terminated in small vascular bundles ang the remaining 28 and 16 were located within large and intermediate vascular bundles respectively. In source leaf tissue 121 of 222 stylets and stylet tracks encountered were associated with small vascular bundles and only 58 tracks and 43 tracks with intermediate and large vascular bundles, respectively. The effect of sustained RWA feeding on the transport capacity was examined after the application of 5,6 carboxyfluoresceine diacetate (5,6-CFDA) in control (sink and source leaf tissue) and aphid-infested (source) wheat leaves, using fluorescence microscopy. After 3h acropetal longitudinal transport of 5,6-CF had occurred in sink leaves in longitudinal veins, as well as a lateral transfer via cross veins and subsequent unloading into mesophyll cells close to the tip of the leaf was observed. In control leaf tissue, the fluorescence front was detected up to about 5cm from the point of application and was only associated with the phloem and not unloaded. In contrast, aphid-infested leaf tissue showed very little 5,6-CF transport, being limited to 2cm or less from the point of application. Structural damage to the phloem in general and to the sieve tubes in particular within of control and infested wheat leaves was investigated using transmission electron microscopy (TEM). In addition, leaf strips were mounted in aniline blue to visualise callose deposition using the fluorescence microscopy. At the TEM level. infested leaf tissue showed various abnormalities, which included destruction of cell contents, membrane damage and subsequent loss of cell contents. TEM studies suggest severe osmotic shock resulted from the aphid's probing. Examination of leaf tissue using fluorescence microscopy showed that there was very little characteristic aniline blue-stained callose visible in control leaf tissue, other than the thin diffuse patches along the sieve plates and punctate spots associated with pore plasmodesmatal areas and plasmodesmatal aggregates. In contrast, the aphid-infested leaf tissue was heavily callosed, with callose deposited not only within the phloem tissue but also in neighbouring vascular parE:}nchyma cells as well. The data collectively suggest that D. noxia feeds preferentially within thin-walled sieve tubes, within the small longitudinal vascular bundles in sink , as well source leaf tissue. Based upon the data presented here the thin-walled sieve tubes in the wheat leaf appear to be more attractive to the aphid and that they are probably more functional in terms of transport system and unlo?lding in sink leaves. Aniline blue stained leaf material that had previously hosted large aphid colonies showed evidence of extensive callose deposits 24 to 36h after the aphids were removed, suggesting that the aphids caused severe mechanical damage to the vascular tissue and mesohyll cells as well. Damage (transient or more permanent) and the subsequent deposition of wound callose, disrupted phloem transport and hence the export of photoassimilate from the leaves.

M.Tech. / Mycotoxins are toxic substances naturally produced by moulds (fungi) that may contaminate agricultural commodities by growing on grain or feed. These moulds produce in a variety of dangerous mycotoxins, such as aflatoxin, fumonisin, deoxynivalenol (DON), ochratoxin, and zearalenone, and may induce acute or chronic effects (carcinogenic, mutagenic, tetragenic and oestrogenic) in humans and animals if ingested. Wheat is an important cereal grown in most provinces of South Africa and yet little is known with respect to incidence of its infection with fungi, contamination with mycotoxins or what carry over of these toxins is into human food. The purpose of this project is to investigate the incidence of fungi and mycotoxins in samples of wheat taken from specified areas in South Africa with a view to finding out if there is indeed a problem and, if so, what would be its impact on the industry and consumers. This pilot project would then form the basis for further studies in other geographical areas and how wheat-based foods for human consumption are affected. Mycotoxins have probably been present in food and feed since the beginning of human existence. Some of their effects have been known for hundreds of years. The technology to detect and chemically characterize them has only really developed in the last 40 years, particularly since 1980. Very small quantities of many of the important mycotoxins can now be detected and accurately measured in foods and feeds. In addition to those already known, many others are known to exist, but have not yet been chemically characterized. Scientists are now identifying toxic compounds in food faster than the information can be processed. However, to maintain perspective, it must be remembered that these substances have always been there, that humans have always been eating the food in which they occur and in the case of many substances, only the dose makes the poison. This study investigated the presence of toxigenic fungi and mycotoxins in wheat and wheat-based products, and also did a pilot study on a few heavily contaminated field wheat (HCFW) samples. Fusarium species dominated the mycological contamination at 40 to 65%, Aspergillus 45%, Penicillium 41% Mucor and Rhizopus were also detected. Among the Fusarium species isolated, six were identified as F graminearum, F. solani and F. vertillioides-like and F subglutinans. Phoma species were also isolated. Quantitative and qualitative analysis by, multi-mycotoxin screen method and VICAM method to analyze foods and feeds was used. Deoxynivalenol was the most prevalent at 45% incidence in samples with a mean concentration of 4.6ppm, aflatoxins in 12% of the samples with a mean concentration of 1.2ppb, ochratoxin in 16% of the samples with a mean of 1.5ppb, zearalenone in 18% of the samples with a mean of 0.1ppb. Attention was drawn to the potential presence of fumonisin in wheat, where fumonitest VICAM method was used, fumonisins were detected on few samples at very low levels of 1 to 2ppm in commercial wheat, and a high reading of 49ppm was recorded on one of the heavily contaminated field wheat (HCFW) samples. Confirmation of these results was carried out with the use of the liquid chromatography coupled with mass spectrum (LC/MS) and high-pressure liquid chromatography (HPLC) system. These results indicate an incidence of mycotoxin in these products, a recommendation of monitoring for prevention of moulds and mycotoxins is suggested. Since health concerns related to dietary exposure to mycotoxins depend on the levels of mycotoxins in the food as consumed, on the amount of food consumed and on the body weight and physiology state of the individual. Therefore, there is a likely possibility that in South Africa, some of the population is exposed to mycotoxins through consumption of wheat and its by-products.

A Thesis Submitted to the Faculty of Science, University of the Witwatersrand, Johannesburg, in fulfillment of the requirements for the Degree of Doctor of Philosophy / The Epidemiology, occurrences and effect of BMV (Brome Mosaic virus) wheat (triticum aestivum) in the summer rainfall area. (Abbreviation abstract) / AC 2018

Call number: LD2668 .T4 1952 H8 / Master of Science

Call number: LD2668 .T4 1986 C374 / Master of Science / Plant Pathology

Photocopy of typescript. / Digitized by Kansas Correctional Industries

ix, 223 leaves : ill., graphs, tables photos (part col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Pathology, 1979

Bibliography: leaves 184-209. This thesis describes the occurence of natural, biologically based suppression of Rhizoctonia barepatch in a direct drilled system at Avon, South Australia. The supressive characteristics are transferable, removed by biocidal treatments, and active against increasing doses of R. solani AG-8, Gaeumannomyces graminis var. tritici and Fusarium graminearum. Disease severity and the viable population of Rhizoctonia are reduced in suppressive soil but the causal agent is still present. The microbial populations in suppressive and non-suppressive soil appear to differ both in their functioning and composition. The control strategy is developed through manipulation of the existing soil biota with farming practices.

Tan spot of wheat is an economically significant disease caused by the fungal pathogen, Pyrenophora tritici-repentis. Certain races of the fungus secrete Ptr ToxA (ToxA), a 13.2 kDa proteinaceous host-selective toxin that is responsible and sufficient to cause disease in susceptible wheat varieties. Disease symptoms develop only when the ToxA gene in the fungus and a single gene in the wheat host are expressed. The understanding of this gene-for-gene interaction could be instrumental towards control of the disease and is also being developed as a model system for understanding host-pathogen interactions. Here, this effort is given a solid structural foundation through crystallographic analysis of the ToxA structure. The ToxA structure was solved at 1.65 Å resolution using the anomalous signal from inherently present sulfur atoms. The monomeric toxin adopts a β-sandwich fold of two anti-parallel β-sheets composed of four strands each. The mapping of existing mutation data onto the structure reveals that a sequence of Arg- Gly-Asp(RGD) and surrounding residues required for activity are present on a solvent-exposed loop thereby making them potential candidates for recognition events that are required for ToxA activity. Unexpectedly, after a simple circular permutation, the ToxA structure is topologically identical to the classic mammalian RGD containing fibronectin type III (FnIII) domain, and furthermore the RGD residues are topologically equivalent. These results support the hypothesis that ToxA, like FnIII, interacts with an integrin-like receptor on the host plant cell surface. There has been a renewed interest in the method of using the anomalous signal from sulfur atoms to solve protein structures. As a spin-off of the structure solution work, the data were systematically analyzed to study the effects of crystal decay, resolution and data redundancy on the ability to locate the sulfur positions and subsequent phasing of the protein. The analyses show that the choices made about data redundancy and resolution limits may be crucial for the structure determination and that anomalous correlation coefficients are helpful indicators in making these choices. / Graduation date: 2006

Cephalosporium stripe of wheat (Triticum aestivum), caused by the soilborne fungus Cephalosporium gramineum, results in significant yield reductions in dryland winter wheat crops of the U.S. Pacific Northwest. The development of resistant cultivars offers the best hope for disease control. Breeding for resistance is hampered by the long trial times inherent in screening adult plants, and by cultivar x environment interactions in field tests. The principal objective of this research was to develop and test a procedure for screening wheat seedlings in controlled environments for resistance to Cephalosporium stripe. Wheat seedlings were raised hydroponically in growth chambers, and the fungus was increased in large fermentation tanks. The seedlings were inoculated at about 12 days post-germination. Disease severity was assessed approximately seven days later using a chlorophyll meter to measure the symptoms of chlorosis and striping. In three trials, five soft white cultivars from the Pacific Northwest and four hard red cultivars from the Southern Great Plains with known levels of field resistance were tested with a Pacific Northwest fungal isolate. With one exception, chlorophyll readings ordered the cultivars appropriately, with moderately resistant cultivars ranking above susceptible cultivars. Three other moderately resistant cultivars from the Pacific Northwest also appeared in one or two trials, and were ranked properly by chlorophyll level. Chlorophyll levels of uninoculated plants were assayed to determine if differences in chlorophyll content were innate in the cultivars. The chlorophyll levels of uninoculated and inoculated seedling treatments were only significantly correlated when the cultivar Madsen, which ranks high both in resistance and in chlorophyll content, was included. In adult plants, flag-leaf chlorophyll level corresponded to intensity of Cephalosporium stripe symptoms where disease was present, and was independent of known field resistance in undiseased cultivars. The seedling screening technique was used to investigate pathogenic variability in C. gramineum. In two experiments, a total of eight cultivars from the Pacific Northwest and the Southern Great Plains were tested with three fungal isolates from each region. No evidence of virulence/vertical resistance was found. There was also no significant adaptation of isolates to greater virulence on cultivars from the same region. / Graduation date: 1998