No description available.

No description available.

No description available.

M.S.

Sclerotinia crown and stem rot (SCSR) incited by Sclerotinia trifoliorum Eriks. causes severe losses in some fall-seeded, no-tillage plantings of alfalfa (Medicago sativa IL.) in Virginia. A mycelial plug inoculation technique was used to detect differences between cultivar (cv) responses of two alfalfa cvs, Arc and Vertus, under greenhouse conditions. A six dia plug from the margin of a 5-day-old culture of S. trifoliorum was placed near the crown area of a plant and incubated for a pre-determined period in a dew chamber at 18 C and 100% RH. Differences in isolate virulence were detected; cv Vertus was less susceptible than Arc to the less virulent isolates while the more virulent isolate (TAL 4) was equally severe on both. An incubation period of 96 hr produced significantly higher disease severity than 72, 48 or 24 hr, however, cv differentiation was best after 72 hr. Eight-, and nine-week-old plants were found to be most suitable for cv evaluation tests since younger seedlings were severely damaged and more mature plants did not develop sufficient symptom expression. Evaluation of twelve cvs with the virulent isolate (TAL 4) and the less virulent isolate (LAL 3) after 96 hr incubation produced significant differences between the mean disease severity ratings (MDSRs). Disease severity increased up to 20 days and then stabilized. Cultivar Anstar followed by WL 320, Vertus and Saranac AR were less susceptible in a majority of the tests; Endure and Euver performed well in some tests while Pioneer Brand 526 and Raidor performed poorly in all tests. This inoculation technique may act as the primary step in the selection of disease resistant germplasm for propagation, re-evaluation, and mass selection before field testing. / Master of Science

M.S.

INTRODUCTION In the summer of 1918, at Blacksburg, Virginia, there developed a considerable amount of a soft rot of tomatoes. This occurred in experimental plots which were designated to study the control of septoria leaf blight, and the soft rot of the fruit developed into an important factor. In describing these experiments Fromme (2) states: "Practically all of the unsoundness of the fruit was caused by bacterial soft rot, a disease which is exceedingly common and often very destructive in tomato fields in Virginia." Isolations from diseased fruits made by S. A. Wingard proved a bacterium to be the causative agent. Its growth in pure culture resembled that of the group of bacteria which causes soft rots of plants but it could not be readily assigned to any of the described species of this group. There has been only casual mention of a bacterial soft rot of tomato in literature, and the distinguishing features of the organisms which might be responsible have not been as sharply defined as is desirable. It was decided, therefore, to undertake comparative studies of the organism in question together with some of the non-chromogenic soft rot forms. / Master of Science

Several potential pathways exist for significant nitrogen loss from nitrogen fertilizer applied to winter wheat in Virginia. A number of materials have been previously identified which inhibit steps in one or more of these loss pathways. Ammonium polyphosphate, ammonium thiosulfate, dicyandiamide, potassium chloride, and monoammonium phosphate were chosen for use in the present study. They were mixed, singly and in combinations, with UAN solution or urea solution and spring top-dressed on soft red winter wheat. Experiments were conducted over two growing seasons at two locations each season. Nitrogen uptake was measured in mid-season and yield measured at harvest. In the second season, field measurement of ammonia volatilization was performed in the experiment utilizing urea solution. Addition of monoammonium phosphate significantly reduced volatilization from urea solution applied to winter wheat at both locations; however, weather conditions were such that total volatilization was low in this season, and the reduced losses were not reflected in increased nitrogen uptake or yield. None of the treatments significantly increased yield in any experiment, nor consistently increased mid-season nitrogen uptake. / Master of Science

Conventional linkage and primary trisomic analyses were undertaken to locate genes conditioning resistance of barley (Hordeum vulgare L.) to Puccinia hordei Otth. The linkage relationships of resistance genes Pa in Speciale (C.I. 7536) and Pa₇ in Cebada Capa (C.I. 6193) with genes known to reside on chromosomes 2 and 3 were studied in crosses involving Speciale and three chromosome 2 markers (e, gs₅, and lr), and in crosses involving Cebada Capa and four chromosome 3 markers (gs₂, uz, als, and yst₂). The location of the resistance gene Pa₃ in Estate was studied by primary trisomic analysis. Inheritance of leaf rust resistance was determined by seedling reaction to culture 57-19, race UN4 of Puccinia hordei. The inheritance and linkage relationships of marker genes and rust resistance were analyzed from F₂ and F₃ coupling data. A recombination value of 41.0 ± 2.2% was found between Pa and gs₅, a marker for chromosome 2L. No evidence was obtained linking Pa with genes e or lr, but population sizes were too small to detect reliably loose linkages in these populations. A loose association was observed between Pa₇ and yst₂ a marker for chromosome 3S; however, the data were not sufficiently homogeneous to permit estimation of a recombination value. The data from crosses involving Pa₇ and uz were not consistent but one population exhibited a trend which suggested these genes might be linked. No evidence of linkage was observed in crosses between Pa₇ and the genes gs₂ and als, although population sizes were limited. The assignment of Pa₇ to chromosome 3 was confirmed by primary trisomic analyses. By the same method, the Pa₃ gene in Estate was shown to be independent of chromosome 3. High temperatures during leaf rust inoculation and incubation periods resulted in poor expression of infection type in crosses involving Estate and Triplo 4, 5, and 6. Despite this the data from these crosses indicated that Pa₃ is not likely inherited through chromosomes 4 through 6. Accrued data suggest that Pa₃ may be located on chromosome 1. / Ph. D.

Barley yellow dwarf virus (BYDV), an aphid transmitted luteovirus, is the most widespread and economically damaging virus of cereal crops. The work in this thesis aims to characterise the basis of the naturally occurring resistance to BYDV in cereals in three ways: Firstly, by facilitating the isolation of the Yd2 gene for BYDV resistance from barley by a map-based approach. Secondly, by determining if a BYDV resistance gene in rice is orthologous to Yd2. Thirdly, by establishing if other BYDV resistance genes in non- Ethiopian barleys are allelic to Yd2. It is hoped that the information generated in this study will ultimately assist in the production of BYDV resistant cereal cultivars. A detailed genetic map of the Yd2 region of barley chromosome 3 was constructed, containing 19 RFLP loci, the centromere and the Yd2 gene. Yd2 mapped on the long arm, 0.5 cM from the centromere, and in the mapping population of 106 F2 individuals, perfectly cosegregated with the RFLP loci XYlp, and Xwg889. This map represents the first stage in a project to isolate the Yd2 gene by a map-based approach. The isolation of Yd2 could help to elucidate the molecular mechanism of the Yd2-mediated BYDV resistance, and may allow the production of BYDV resistant cereals by genetic transformation. The RFLP markers mapped closest to Yd2 could also be useful in barley breeding, by enabling selection for both the presence of Yd2 and the absence of agronomically undesirable traits known to be closely linked to Yd2. Genetically Directed Representational Difference Analysis (GDRDA) is a technique based on subtractive hybridisation, which can be used to identify RFLP markers closely linked to a gene of interest. Two GDRDA experiments were performed with the intention of generating additional RFLP markers close to Yd2. However, the first experiment yielded RFLP probes that were not derived from the barley genome, while the second experiment yielded probes that detected repetitive sequences. It was concluded that GDRDA is of limited use in generating further markers close to Yd2. To isolate the Yd2 gene by a map-based approach, a much larger mapping population will need to be analysed to genetically resolve markers tightly linked to Yd2. If the two morphological markers uzu dwarf and white stripe,,j flank Yd2, then they could assist in this task by enabling the visual identification of F2 seedlings resulting from recombination close to Yd2. However, in this study, both morphological markers were found to be located distal to Yd2. Therefore, these two morphological markers can not be used together to facilitate high resolution genetic mapping of the Yd2 locus. It may be possible to use large-insert genomic DNA clones from the relatively small genome of rice to generate further RFLP markers close to the Yd2 gene in barley, provided that the order of orthologous sequences in barley and rice is conserved close to the Yd2 locus. To assess the feasibility of this approach, RFLP probes used to identify loci close to Yd2 were mapped in rice using a segregating rice F2 population. Five of the RFLP loci mapped together and in the same order as RFLP loci mapped close to Yd2 in barley using the same probes. By comparing the location of RFLPs mapped by other researchers in rice using probes mapped close to Yd2, the region of conserved linkage between rice and the Yd2 region was tentatively identified as the central portion of rice chromosome 1. The collinearity shown by orthologous sequences in barley and rice indicated that it may indeed be possible to use rice to assist in generating RFLP markers close to Yd2. Of all the cereals, rice is the most amenable to map-based gene isolation, due to its small genome, well developed physical and genetic maps, and its ability to be genetically transformed with high efficiency. If a BYDV resistance gene that is orthologous to Yd2 could be identified in rice, this gene could be isolated with relative ease, and then used to identify barley cDNA clones corresponding to Yd2 gene by virtue of the sequence homology expected between these genes. To test if a BYDV resistance gene from an Italian rice line is orthologous to Yd2, recombinant-inbred rice lines previously characterised for this gene were analysed using probes mapped close to Yd2 in barley. No genetic linkage was detected between the RFLP loci and the BYDV resistance gene, indicating that the gene is unlikely to be orthologous to Yd2. BYDV resistance alleles at the Yd2 locus which are of a non-Ethiopian origin may show interesting differences to Ethiopian Yd2 resistance alleles. To identify barleys which may contain resistance alleles of Yd2, ten BYDV resistant barleys not known to contain Yd2 were assessed for their resistance to the PAVadel isolate of BYDV in the glasshouse. CI 1179, Rojo, Perry, Hannchen, Post and CI 4228 were found to be the most resistant under these conditions, and were analysed further. If the resistance from these barleys is controlled by alleles of Yd2, RFLP markers close to Yd2 will be expected to cosegregate with the resistance in F2 families derived from crosses between these resistant barleys and the BYDV susceptible barleys Atlas and Proctor. RFLPs suitable for use in these allelism tests were identified using probes mapped close to Yd2. However, time did not permit the analysis of these F2 populations. / Thesis (Ph.D.) -- University of Adelaide, Dept. of Plant Science, 1996