Thesis (M.S.)--Oregon State University, 2008. / Printout. Includes bibliographical references (leaves 70-78). Also available on the World Wide Web.

Grapevine powdery mildew is caused by Erysiphe necator, an Ascomycete fungus and an obligate biotroph restricted to growth on its grapevine host. Biotrophic pathogens form a stable association with host cells without directly causing cell death, and take up nutrients from, in the case of powdery mildew ( PM ), host epidermal cells ( Rumbolz et al., 2000 ). As the fungus grows, its increasing biomass becomes a strong nutrient sink capable of altering assimilate flow and storage in the host. To identify host genes that may mediate nutrient delivery to powdery mildew infected tissues and therefore may contribute to disease susceptibility, a candidate gene approach using degenerate and RT - PCR, and a nontargeted approach using microarray analysis was instigated. Once identified, " susceptibility genes " could be targeted for manipulation to provide alternative resistance strategies based on reduced susceptibility in the future. In addition to genes encoding pathogenesis and stress related proteins, microarray analysis revealed that transcript levels of a putative metal transporter and a cell wall structural protein were elevated in infected berry skin, while aquaporin water channels and genes associated with photosynthesis were generally repressed. Degenerate PCR was used to isolated new cell wall invertase, monosaccharide and amino acid transporter genes and initial RT - PCR revealed that expression of genes involved in sugar mobilisation were the most significantly modulated by powdery mildew infection. Previously unreported hexose transporters ( HTs ), ( VvHT3, VvHT4 and VvHT5 ) and a cwINV ( VvcwINV ) had been isolated from cDNA prepared from powdery mildew infected grapevine leaves. Full length clones of grapevine HTs and cwINV were obtained by RACE PCR. Heterologous expression of the three new HTs in yeast confirmed that VvHT4 and VvHT5 mediated glucose uptake, while VvHT3 did not function in the yeast system. However, transient expression of a translational fusion of the VvHT3 protein with green florescence protein in onion epidermal cells indicated that it is targeted to the plasma membrane of plant cells. Quantitative RT - PCR analysis of these new genes, together with previously reported grapevine HTs and cytoplasmic and vacuolar invertases, indicated that expression of VvcwINV and VvHT5, were significantly up - regulated by PM infection, while a vacuolar invertase was strongly down - regulated by PM infection. Invertase activity assays were in agreement with these findings, showing elevated sucrolytic activity in insoluble fractions and reduced sucrolytic activity in soluble fractions. These results suggest that apoplasmic phloem unloading of sucrose in the infected leaf is elevated and that VvHT5 is induced to recover the additional hexoses from the apoplasm. Basic localisation studies indicated that VvHT5 and VvcwINV are not induced specifically in powdery mildew infected leaf regions, but are induced in a more diffuse distribution within infected leaves. To determine if induction of VvHT5 and VvcwINV is specific to PM infection or if other stimuli may also mediate these responses, leaves were inoculated with downy mildew or stressed by wounding. Transcript levels of VvHT5 and VvcwINV were elevated by wounding and downy mildew infection, suggesting that the induction of these genes may be part of a general stress response. To explore the signalling pathways that may underlie these responses, leaves were treated with the plant growth regulators ethylene, jasmonate and abscisic acid. Exogenous application of ethylene and methyl jasmonate only marginally affected the expression of the genes studied, however foliar application of abscisic acid ( ABA ) induced gene expression changes similar to those observed in response to powdery mildew infection and wounding. Promoter sequences of VvHT3, VvHT4, VvHT5 and VvcwINV were isolated and analysed for the presence of regulatory elements. Compared with the promoters of VvHTs that were not induced by pathogen infection or wounding, the VvHT5 and VvcwINV promoters contained numerous motifs associated with induction by ABA including ABRE, Myc and Myb binding elements. The path of sugar loading into the mesocarp of grape berries during ripening is still poorly understood and few molecular components associated with this process have been described. Quantitative RT - PCR was used to monitor the expression of five HTs and VvcwINV during Cabernet sauvignon and Shiraz berry development and ripening. Of the three new HTs reported here, the expression of VvHT3 is most consistent with a potential role in sugar loading, while VvHT5 is induced late in this process. VvcwINV transcript levels were high pre - ripening and also during the later stages of ripening, therefore based on this expression pattern, a role for this enzyme during ripening is not clearly evident. These results are discussed in terms of an apoplasmic step in phloem unloading in ripening grape berries. This study has provided new insights into the molecular and biochemical processes associated with the formation of carbohydrate sink metabolism in response to stress stimuli, and sugar delivery to grape berries during ripening. ABA - dependant pathways may mediate the stress - associated induction of VvcwINV and VvHT5, presumably to recruit additional carbohydrates to the affected organ to energise repair and defence responses. At this stage it is unknown if this response is beneficial to pathogen nutrition, however potentially, modification of genes associated with carbohydrate sink metabolism could provide an alternative way to engineer resistance to this pathogen. / Thesis (Ph.D.)--School of Agriculture, Food and Wine, 2006.

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Sorghum downy mildew (SDM) of maize is caused by the oomycete Peronosclerospora sorghi (Weston and Uppal) C. G. Shaw. The disease can cause devastating yield losses in maize (Zea mays L.). Quantitative trait loci (QTLs) mediating resistance to SDM were mapped using both restriction fragment length polymorphisms (RFLPs), and simple sequence repeats (SSRs) in 220 F2 individual maize progeny derived from a cross between two extremes; highly susceptible inbred parent SC-TEP5-19-1-3-1-4-1-1 (white) and highly resistant inbred P345C4S2B46-2-2-1-2-B-B-B (yellow). The phenotypic expression was assessed on F2:3 families in a wide range of environments under natural field infection and in a controlled greenhouse screening method. Heritability estimates of disease reaction ranged from 93.3% in Thailand sit 1 to 48% in Thailand sit 2. One hundred and thirty three polymorphic markers were assigned to the ten chromosomes of maize with LOD scores exceeding 4.9 covering about 1265 cM with an average interval length between markers of 9.5 cM. About 90% of the genome was located within a 10 cM distance to the nearest marker. Three putative QTLs were detected in association with resistance to SDM in different environments using composite interval mapping. Despite environmental and symptom differences, one QTL on chromosome 2 bin 9 had a major effect in all trials and explained up to 70% of the phenotypic variation in Thailand where the highest disease pressure was experienced. Two other QTLs on chromosome 3 bin 5 and chromosome 9 bin 2 had a minor effect, each explaining no more than 4% of the phenotypic variation. These results revealed one major gene and two minor genes that control sorghum downy mildew resistance. These markers should be very useful in breeding programs in facilitating the introgression of the resistance genes into commercial varieties. Marker-assisted selection for these loci should be useful in incorporating SDM resistance genes in maize across environments, even in the absence of the pathogen.

4 pp. / Mesquite (Prosopis sp.) is a popular tree in landscapes in Arizona because of its drought tolerance and attractive growth habit. Powdery mildew has been observed from late summer until early spring on mesquite leaves. It has been identified as Pleochaeta polychaeta based on morphological descriptions and comparison to herbarium specimens. Surveys were conducted in fall 2008 through winter 2009 at two locations in southern Arizona to determine the susceptibility of different mesquite species to powdery mildew. Twelve mesquite trees representing two species were sampled at Texas Canyon near Willcox, AZ, and 177 trees representing eight species were sampled at the University of Arizona campus in Tucson, AZ. The North American mesquite species P. glandulosa var. glandulosa and P. velutina were infected with powdery mildew at the University of Arizona campus and P. velutina at the Texas Canyon site. No powdery mildew was observed on P. alba, P. cinerea, P. nigra, P. chilensis, P. pubescens and P. chilensis x flexuosa. The powdery mildew affects the aesthetic value of severely infected trees but seems to have little effect on long term tree health.

Sorghum downy mildew (SDM) of maize is caused by the oomycete Peronosclerospora sorghi (Weston and Uppal) C. G. Shaw. The disease can cause devastating yield losses in maize (Zea mays L.). Quantitative trait loci (QTLs) mediating resistance to SDM were mapped using both restriction fragment length polymorphisms (RFLPs), and simple sequence repeats (SSRs) in 220 F2 individual maize progeny derived from a cross between two extremes; highly susceptible inbred parent SC-TEP5-19-1-3-1-4-1-1 (white) and highly resistant inbred P345C4S2B46-2-2-1-2-B-B-B (yellow). The phenotypic expression was assessed on F2:3 families in a wide range of environments under natural field infection and in a controlled greenhouse screening method. Heritability estimates of disease reaction ranged from 93.3% in Thailand sit 1 to 48% in Thailand sit 2. One hundred and thirty three polymorphic markers were assigned to the ten chromosomes of maize with LOD scores exceeding 4.9 covering about 1265 cM with an average interval length between markers of 9.5 cM. About 90% of the genome was located within a 10 cM distance to the nearest marker. Three putative QTLs were detected in association with resistance to SDM in different environments using composite interval mapping. Despite environmental and symptom differences, one QTL on chromosome 2 bin 9 had a major effect in all trials and explained up to 70% of the phenotypic variation in Thailand where the highest disease pressure was experienced. Two other QTLs on chromosome 3 bin 5 and chromosome 9 bin 2 had a minor effect, each explaining no more than 4% of the phenotypic variation. These results revealed one major gene and two minor genes that control sorghum downy mildew resistance. These markers should be very useful in breeding programs in facilitating the introgression of the resistance genes into commercial varieties. Marker-assisted selection for these loci should be useful in incorporating SDM resistance genes in maize across environments, even in the absence of the pathogen.