Fusarium wilt of lettuce was first recognized in Arizona in 2001. Since this first discovery, the pathogen, Fusarium oxysporum f.sp. lactucae (Fol), has been recovered from infected lettuce plants from approximately 30 different fields. This fungus is a soil-borne pathogen that can remain viable in soil for many years. Cultural disease control measures, such as extended soil flooding and soil solarization, have shown promise in managing Fusarium wilt in other cropping systems. The specific research objective during the 2005 growing season was to further evaluate the effect of preplant solarization of planting beds on subsequent development of Fusarium wilt on lettuce. There was no significant difference between the short (28 days) and long (56 days) solarization period in the subsequent number of diseased lettuce plants; therefore, the disease incidence values for both solarization periods were combined and compared to nonsolarized plots. At each data collection date, the number of lettuce plants showing symptoms of Fusarium wilt was significantly lower in solarized beds compared to nonsolarized beds. At plant maturity (Nov 18), Fusarium wilt had claimed virtually all lettuce plants of the cultivar 'Lighthouse' growing in nonsolarized soil; however, only 19% of lettuce plants of the same cultivar growing in solarized soil showed disease symptoms. This equates to an 81% reduction in diseased plants in solarized soil compared to nonsolarized soil. The results of this field trial suggest that a 30-day summer solarization treatment of lettuce beds can significantly reduce the inoculum of Fusarium oxysporum f. sp. lactucae to levels that would allow substantial growth of a susceptible lettuce cultivar. Additional field studies are needed to refine the solarization process to potentially achieve further increases in efficiency of destroying propagules of Fusarium oxysporum f. sp. lactucae in infested fields.

Powdery mildew occurs annually on melons in Arizona. Podosphaera xanthii (Sphaerotheca fuliginea) is the plant pathogenic fungus that causes powdery mildew on cucurbits, such as cantaloupe, honeydew, watermelon, cucumber and squash. Development of powdery mildew on melons is favored by moderate temperatures and relative humidity, succulent plant growth and reduced light intensity brought about by a dense plant canopy. Existing products as well as materials under development were evaluated and compared for efficacy in management of powdery mildew on cantaloupe in a field trial conducted during the spring of 2007 at the University of Arizona, Yuma Valley Agricultural Center. A moderately-high level of disease had developed when disease severity data was recorded at crop maturity in mid-June. Among treatments, the degree of powdery mildew control ranged from essentially 0 to 100%. Materials that reduced the severity of disease on both the top and bottom of leaves by an average amount of at least 90% included Microthiol Disperss, Procure, Quintec, V-10118, Inspire Super, Endura, Cabrio, and Pristine, when applied on a 7-day spray interval. Other treatments that reduced powdery mildew by at least 80% compared to untreated plants included alternation between the conventional fungicide Procure and the biopesticides, Serenade Max, Sonata, or Actinovate as well as alternation among the two biopesticides Actinovate and Kaligreen. Alternating applications of products with different modes of action is imperative to minimize the development of insensitivity in the pathogen population to one or more of these active ingredients. Among the products evaluated this year, several are registered for use in Arizona for control of powdery mildew on melons.

Fusarium wilt of lettuce was first recognized in Arizona in 2001. Since this initial discovery, the pathogen, Fusarium oxysporum f.sp. lactucae (Fol), has been recovered from infected lettuce plants from over 40 different fields. This fungus is a soil-borne pathogen that can remain viable in soil for many years. Soil solarization has shown promise in managing Fusarium wilt in other cropping systems as well as in lettuce field trials conducted during 2004 and 2005. In an effort to maximize the solarization effect on subsequent disease development, the following factors were evaluated in a 2006 field trial: 1) solarization of unshaped versus preshaped beds, 2) the effect of soil moisture on solarization efficiency of preshaped beds, and 3) effect of lettuce type on Fusarium wilt incidence after solarization. The entire field was flood irrigated on Jun 21. Plots were solarized during Jul and/or Aug by covering beds with 1-mil thick clear plastic. During the solarization treatment from Jul 3 to Sep 10, the mean soil temperature in preshaped solarized beds at a depth of 2 and 9 inches was 116 and 95°F, respectively, and 102 and 97°F, respectively, in beds not covered with plastic. When solarization was initiated 15 days after soil irrigation, a 20% reduction in Fusarium wilt was recorded for a crisphead lettuce cultivar grown on solarized unshaped beds compared to a 56% reduction in disease when the same crisphead cultivar was grown on preshaped solarized beds. There was no significant difference between a one and two month solarization period in the subsequent number of diseased lettuce plants. Solarization of preshaped beds 15 days after irrigation for one month reduced Fusarium wilt on crisphead lettuce by 56%, whereas the same solarization period initiated seven days after irrigation resulted in a 96% reduction of disease. The same one-month solarization period started one week after soil irrigation reduced the incidence of Fusarium wilt on green leaf (Two Star) and romaine (Green Towers) by 97 and 88%, respectively, compared to plants grown on unsolarized beds. The data show that summer solarization of moist soil can 1) destroy propagules of Fusarium oxysporum f. sp. lactucae in infested fields and 2) be a useful cultural management tool to significantly reduce the incidence of Fusarium wilt in a subsequent crop of lettuce.

Gray mold, caused by the fungus Botrytis cinerea, usually has been considered a minor disease in field-grown lettuce. However, for the past two lettuce production seasons, the incidence of this disease has increased significantly in Southwestern Arizona lettuce fields. Most of the infected fields were planted to romaine lettuce; however, some iceberg lettuce plantings were involved as well. The occurrence of gray mold was most prevalent during February and March. The primary symptoms of the disease include plant wilting accompanied by a fuzzy gray growth at the plant base, which contains masses of airborne spores. Sclerotia may also be present on infected tissue. Infected plants that show these symptoms usually wilt and die. Botrytis cinerea can survive on crop debris, as a pathogen on numerous crops and weed hosts, and as sclerotia in soil. Airborne spores that land on senescent or damaged lettuce stems and leaves germinate and rapidly colonize this tissue. Once established, the pathogen grows into adjacent healthy stems and leaves. A field trial was established to 1) evaluate the effectiveness of some fungicides to reduce the incidence of Botrytis gray mold and 2) compare the susceptibility of some lettuce cultivars to this disease. Botrytis gray mold did not develop in plots of the crisphead cultivar 'Winterhaven'; however, the disease was present in all plots planted with cultivars of romaine lettuce. Among the tested romaine cultivars, Botrytis gray mold incidence was lowest in 'A 35585-1', 'Fresh Heart' and 'Rome 59' in untreated plots as well as in plots treated four times with the fungicides Rovral, Endura, or Switch. On the other hand, disease incidence was highest on the cultivar 'Green Towers' in untreated plots as well as those treated with one of the three tested fungicides. To compare the ability of tested fungicides to control Botrytis gray mold, the disease incidence values for each of the five romaine cultivars treated with each specific fungicide were pooled together and compared to disease levels recorded on untreated plants. Foliar applications of treatments were made Jan 24, Feb 1, Feb 16 and Feb 28, 2007. A 31% reduction in disease was recorded on romaine lettuce plants treated with Switch, whereas a 17% reduction in Botrytis gray mold was observed on plants treated with Endura. Rovral did not significantly reduce Botrytis gray mold in this trial. Additional field trials are planned to verify these initial findings.

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Oxalate oxidase is a water soluble, thermolabile, homo-oligomeric glycoprotein the synthesis of which marks the onset of germination In wheat and barley embryos. The protein Is also highly abundant In barley roots. The enzyme has an average oligomer molecular mass of about 115 kDa and about 22.8 kDa for the monomers, as determined by mass spectrometry. The ollgomeric cereal oxalate oxidases are resistant to dissociation In SDS containing media and to digestion by pepsin. The cereal organs produce two oxalate oxidase Isoforms (G and G') which possess the same apoprotein but are differentially glycosylated. The oligosaccharide side chain(s) has a molecular mass of about 2-3 kDa. Barley root also contains a third active oxalate oxidase isoform with a mass of about 22.5 kDa, which was not detected in germinating embryos of the same cultlvar. All of the cereal oxalate oxidases were shown to have identical N-terminal amino acid sequences and almost identical kinetic properties This thesis describes the characterisation of oxalate oxidases Isolated from three transgenic plants lines, expressing chimeric CaMV 35S-oxalate oxidase genes. SGS5 tobacco was expressing a gene with the native oxalate oxidase signal peptide and 3S1 oilseed rape and C26 tobacco were expressing a gene containing a foreign extensin signal peptide. Transgenic SGS5 tobacco produced an oxalate oxidase which was almost indistinguishable from the native cereal protein, in terms of Its structure, stability, enzyme activity and resistance to dissociation In SDS containing media and digestion by pepsin. This work Illustrated the ability of a dicotyledonous plant (tobacco) to recognised and correctly process a transgenic monocotyledon protein (wheat).Transgenic 3S1 oilseed rape and C26 tobacco were shown to produce active oligomeric oxalate oxidases, which did not exhibit any of the unusual resistance properties normally associated with these proteins. Instead the 3S1 and C26 oxalate oxidases were unstable and exhibited significantly altered kinetic properties compared with the native cereal and transgenic SGS5 enzymes. The instability was thought to have arisen from the Incorrect processing of the 3S1 and C26 oxalate oxidases, resulting in the partial cleavage of the extensin signal peptide, which in turn gave rise to a mature oxalate oxidase with an altered N- terminal sequence compared with the native cereal enzyme. The use of vacuum infiltration confirmed the association of the transgenic enzymes with the extracellular spaces, although the majority of the enzyme was shown to be intracellular. The main objective for producing the transgenic oilseed rape expressing oxalate oxidase was to Improve fungal pathogen resistance against oxalic acid secreting pathogens. The results described in this thesis are concerned with a direct comparison of the structure, stability and kinetics between the native cereal and transgenic oxalate oxidases and the possible consequences for pathogen resistance In plants expressing unstable yet active transgenic enzymes.

Faculty of Science School of Pathology 9903547r robyn.brakin@gmail.com / Mycobacterium tuberculosis is an important human pathogen, claiming more lives per annum than any other single infectious organism. The host environment of M. tuberculosis contains DNA-damaging agents that pose a constant threat to the M. tuberculosis genome, and as a result, the ability to repair damaged DNA is likely to play an important role in bacterial survival. Y-family polymerases perform translesional synthesis and replicate DNA in an error-prone manner. By characterising the Y-family polymerases in mycobacteria, a better understanding the organism’s adaptive mutagenesis may be established. Through gene expression studies, it was found that UV irradiation of Mycobacterium smegmatis resulted in the up-regulation of dinP3, which was determined to be a Y-family polymerase by sequence analysis. DinP3 expression was found to be under control of the SOS response and is the first example of a Y-family polymerase in mycobacteria forming part of the SOS regulon. However, loss of DinP3 did not change the ability of M. smegmatis to tolerate UV irradiation. Mutagenesis studies revealed a complex interaction between the different Y-family polymerases in M. smegmatis. It was shown that spontaneous mutagenesis was increased in the absence of DinP3, whereas UV-targeted mutagenesis was increased in the absence of DinP, another Y-family polymerase. In conclusion, these results reflect the differences in control and in the mutational profiles of the Y-family polymerases in M. smegmatis. Moreover, these polymerases exhibit distinctive features from other bacterial Y-family polymerases, highlighting the different way in which bacteria have adapted to deal with lesions in their genetic material.