Forage production and longevity of alfalfa (Medicago sativa L.) are reduced by the effect of the fungus Phytophthora megasperma Drechs. in irrigated alfalfa fields located in Central Mexico. There are no resistant alfalfa cultivars adapted to this region. Phytophthora root rot was found in irrigated fields at elevations between 1600 to 2000 m. Strains of this fungus isolated from soil collected in Mexico were more virulent than one strain from Arizona. Four alfalfa cultivars of Mexican origin, Certified Hayden, and Hayden PX-III, an experimental cultivar resistant to strains of P. megasperma from Arizona, were susceptible to P. megasperma Bajio strain collected in Central Mexico. Following two cycles of phenotypic current selection, the level of resistance to Phytophthora root rot was increased from 5.6% to 22.8% in the new population originating from the cultivar INIA-76, and from 12.8% to 30.6% in the new population originating from the cultivar Hayden PX-III.

Resistance (R) genes, a key factor in determining the resistance of plants, have been shown often to be highly allelic entities existing in duplicated regions of the genome. This characteristic suggests that R-gene acquisition may have arisen through frequent genetic rearrangements as a result of transient, reduced genome stability. Tabacco plants transgenic for a recombination construct exhibited reduced genome stability upon infection with a virulent pathogen (tobacco mosaic virus). The reduced genome stability manifested as an increase in recombination events in the transgene. Such increases were observed following a virulent pathogen attack. This increase in recombination was shown to be systemic and was observed prior to systemic viral movement suggesting the presence of a systemic recombination signal. Further molecular analyses revealed that specific R-gene loci experience a large frequency of rearrangements following a virulent pathogen encounter. The possible targeting of instability to R-gene regions may be controlled through epigenetic processes, in particular, DNA methylation. / xiii, 119 leaves ; 29 cm.

Plant-wide communication between tissues and cells is organized, in part, by a suite of compounds called hormones. I have chosen to focus on the effects of one plant hormone, ethylene; how its synthesis is controlled and how its perception is mediated to differentially control cell development and response to pathogens. In the production of ethylene, one level of control is by modulating the levels of the immediate precursor to ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC). I characterize here a plant encoded gene homologous to bacterial ACC Deaminases, AtACD1, and show through up- and down-regulation of the gene that it can modulate the plants sensitivity to exogenous ACC. Once ethylene is produced, it is sensed in Arabidopsis thaliana by a family of 5 receptors. I show that ETR2 in Arabidopsis is responsible for modulation of the microtubule cytoskeleton assembly as loss-of –function mutations to this gene cause randomized microtubule assembly in trichomes and increase sensitivity to microtubule depolymerising drugs in root hairs. In studies of plant:pathogen interactions, ethylene is a central signaling agent required for plant resistance. While it has been shown that etr1 mutants show increased susceptibility to fungal pathogens, exogenous ethylene has also been shown to speed the progress of pathogenesis. Using Fumonisin B1 (FB1) to induce cell death I show that etr1-1 has accelerated cell death while ein4-1 has a reduced rate of necrosis. Further to this, mutations to the other three ethylene receptors do not have any effect on the rate of cell death. My interest in cell development led to the characterization of an activation tagged Populus tremula x P. alba line with increased trichome initiation. The gene responsible for these phenotypes was identified as PtMYB186, which also affected growth rate, transpiration rate, photosynthetic capacity, and resistance to the Tussock moth larvae. Together these studies provide a new framework for our understanding of how the ethylene signal is modulated in plants and the controls behind cellular development. This knowledge will help reconcile studies which show that ethylene has different effects on plant development and provide new avenues of research into trichome development. / Thesis (Ph.D, Biology) -- Queen's University, 2009-01-13 10:08:03.605

The number of lesions increased with increasing temperatures over the range of wet periods except at 30$ sp circ$C, where their number decreased with increasing wetness duration. Cultivars were evaluated for partial resistance under field and greenhouse conditions. In the field ranking was based on cluster analysis of the standard area under the disease progress curve (SAUDPC) for intervals between sampling dates. In the greenhouse, the cultivars were evaluated on the basis of their response relative to five components of partial resistance: the SAUDPC, mean lesion area (MLA), pycnidial density (PCD), spore density (SPD), and the latency period defined as the time from inoculation to 50% and 75% disease (T$ sb{50}$ and T$ sb{75}$). In the greenhouse, overall ranking was based on cluster and principal component analysis of responses to SAUDPC, MLA, PCD and SPD. T$ sb{50}$ and T$ sb{75}$ were not significant. Three cultivars, Golden Plume, Superdora and Summit, were rated as moderately resistant in both field and greenhouse trials. The others ranged from moderately susceptible to very susceptible. (Abstract shortened by UMI.)

In this study, we have demonstrated that inoculation of bean seeds with non-pathogenic binucleate Rhizoctonia (np-BNR) at sowing protected bean seedlings from infection of R. solani. Using quantitative real-time RT-PCR (QRT-PCR), transcript levels of defense genes encoding 1,3-beta-glucanase (GLUC), phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) in one-week old bean seedlings was monitored during np-BNR and R. solani interaction. The results revealed that protection effect of np-BNR correspond to a systemic suppression of these three defense genes' expression from significant higher level elicited by R. solani to the level of non-infected plants. This indicates that bio-protection by np-BNR isolates is not correlated to activation of these three defense genes' expression. Similar suppression was achieved for pre-colonization of bean seedlings with arbuscular mycorrhizal (AM) Glomus introradices on GLUC gene expression, although the AM fungus did not significantly reduce rot symptoms. Possible mechanisms implicated in down-regulation during plant-pathogen and np-BNR or AM interaction are discussed.

Field studies of a population of Arisaema triphyllum affected by the systemic rust Uromyces ari-triphylli were conducted to examine dynamics of disease transmission and to identify factors influencing plant susceptibility to rust infection. Of undiseased plants that developed foliar infections during the growing season, 55% possessed active resistance to the rust disease, and were able to prevent the foliar infection from becoming a systemic one. Flowering increased the susceptibility of plants to both Mar and systemic infections. A phenotypic selection study was carried out to estimate selection on five plant traits and to determine the role of foliar infections of U. ari-triphylli in mediating selection. Multivariate path analysis suggests that both leaf diameter and a general size factor are under positive directional selection. Pathogen-mediated selection coefficients are small, but comparable in magnitude to overall selection coefficients (15%, on average), indicating the potential importance of diseases as selective agents in nature.

Previous studies on Leafy reduced-stature (LRS) maize found that it had extremely early maturity and a higher harvest index (HI), leading to high yields for its maturity rating. Whether this apparent high HI is relaxed to its earliness, or can also exist among the medium or late maturity LRS maize has not been previously investigated. It was also of interest to know if the traits that produced the LRS canopy structure have pleiotropic effects on root architecture. Finally, field observations indicated that LRS maize had a lower incidence of common smut. It is not known whether this apparent resistance is specific to smut or includes other diseases. / Using a wide range of the most recently developed LRS hybrids and some conventional hybrids, a two-year field experiment was conducted to examine the HI and disease resistance of LRS maize. HI, yield, and yield components were compared between the two genotype groups (LRS and conventional) under different population densities. The resistance to the natural incidence of common smut and artificially inoculated Gibberella ear rot was also tested. Morphology and fractal dimension analyses of roots at an early development stage were conducted in indoor experiments. These analyses were performed with WinRHIZO (version 3.9), an interactive scanner-based image analysis system. / This work showed that: (1) There was no relationship between the HI and maturity; higher HIs can also exist among the medium and late maturity LRS hybrids. (2) While LRS maize hybrids have the potential for high yield this was not realized in the LRS hybrids used in this work. Further breeding and development of optimum management practices are needed to fully exploit this potential. (3) During early development LRS hybrids generally had more branching and more complex root systems than conventional hybrids. (4) Fractal dimension, as a comprehensive estimation of root complexity, was highly related to major root morphological variables, such as root total length, surface area, branching frequency and dry mass. (5) Of the hybrids tested the greatest resistance to both common smut and Gibberella ear rot, two major ear diseases, occurred in some of the LRS types.

Meloidogyne javanica is the most widely spread nematode pest on soybean in South Africa. Only a few registered cultivars have some resistance to this nematode and there is an urgent need for an efficient breeding programme for resistant cultivars of all maturity groups. However, breeding is hampered by laborious screening procedures for selection of resistant lines. The objective of this study was to develop an economically viable molecular marker system for application in selection procedures. Three techniques of marker identification were investigated, i.e. RAPD, RFLP and AFLP analysis. The RAPD technique proved to be applicable in fingerprinting soybean varieties, but was too sensitive for interplant variation to be used as an efficient system for identification of molecular markers linked to nematode resistance. Both RFLP and AFLP screening identified markers linked to gall index variation in a segregating population of 60 F₂ progeny from across between a resistant cultivar, Gazelle, and a highly susceptible variety, Prima. A codominant RFLP marker( B212) was linked significantly to resistance and explained 62% of the variation in gall index. Seven AFLP markers were linked significantly to the resistance trait, of which four were linked in repulsion phase and three in coupling phase. All seven AFLP markers mapped to LG-F on the public soybean molecular map. The QTL for resistance mapped between markers E-ACC/M-CTC2 linked in coupling phase, 8212 and E-AAC/M-CAT1, linked in repulsion phase. These two AFLP markers bracketing the major resistance QTL were successfully converted to SCARs. Marker E-ACC/M-CTC2 was converted to a codominant SCAR marker SOJA6, which acounted for 41% of variation in gall index in the mapping population. Marker E-AAC/M-CAT1 was converted to a dominant SCAR marker (SOJA7) and explained 42% of gall index variation in the mapping population. These two markers mapped approximately 3.8 cM and 2.4 cM respectively from the resistance QTL. This study represents the first report of the development of PCR-based sequence specific markers linked to resistance to M. javanica in soybean. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2000.

The complexity of the behaviour of neonate Eldana saccharina Walker (Lepidoptera: Pyralidae) larvae and the limited information on their response to the morphological characteristics of South African sugarcane varieties was the primary justification to study antixenotic/antibiotic effects on larval behaviour. Laboratory experiments were conducted with stalk segments in plastic jars inoculated with larvae and in a metal cage covered with gauze. In jars, the larvae were observed until they penetrated the stalks. After 14 days, the stalks were dissected and larvae weighed. In all varieties, larvae moved directly to the node after inoculation and penetrated the stalk through leaf scars and buds. No significant differences in larval mass were observed among varieties. In cage experiments different parts of the node, namely the rind below the wax band; the bud; and the root primordia were tested. There was a clear indication that rind hardness and the budscale properties are associated with varietal resistance and only affect early instars. The experiments were repeated using whole cane plants in a glasshouse. The results were similar to those of laboratory experiments. In the Insect Rearing Unit, scraped waxes from different varieties were incorporated into the diet. Larval masses from different diets showed significant differences among varieties, but they did not conform to the known resistance ratings, as cane varieties N12 and N21 showed high susceptibility, instead of resistance. Dispersal behaviour of neonates shortly after hatching was investigated in 'mobility experiments' conducted on live cane plants. Mobility is important because the more time neonates spend wandering around on the stalk surface or on exposed parts of the plant, the more vulnerable they are to predation and other adverse factors that may reduce their survival. Experiments to test stalk penetration by larvae on the node showed that neonates required a softer food source before attacking the hard nodal parts. Second and third instar larvae were used subsequently to the mortality of all neonates fed on the rind, which in turn resulted in non-significant differences, suggesting that feeding on debris and/or leaves is critical to the survival and penetration of larvae into the sugarcane stalk. Incorporation of the characteristics tested in these experiments aims to reduce the number of larvae that penetrate the stalk and to expose them for longer on the surface where their numbers may be controlled by predators and insecticides. The resistant varieties used in these experiments have high fibre and less sugar, but newer varieties, such as N29 and N33 incorporate both high resistance and high sucrose yield, which are the two key elements for optimised sugar production. Chemical characteristics of the plants need to be taken into consideration as high sucrose is seldom found in fibrous varieties. Leaf sheath tightness is another characteristic that would go well with leaf sheath hairiness, because though not tested in this work-would make it difficult for the larvae to get to the smooth adaxial surface of the leaf. The hardness of trichomes is another feature that needs to be investigated, because a variety may have dense, but soft pubescence that does not repel even the most sensitive larvae, neonates. At present, integrating plant resistance with cultural control, i.e. field hygiene etc. is cost-beneficial for the sugar industry. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2002.

The Wheeler Orchid Collection is home to some of the most endangered species of orchids in the world. This fantastic reservoir of endangered species has been enhanced and broadened by its function as a plant rescue station for the U.S. customs service. Unfortunately, this responsibility increases the risk of bringing orchids, which harbor contageous diseases, into the greenhouse where sap transmitted diseases such as the Tobacco Mosaic Virus (TMV), can run rampant. Although manipulation of orchid characteristics is typically done by classical plant breeding techniques, genetic engineering is emerging as a useful technique for the introduction of desirable traits into the orchid genome. Through the use of genetic engineering techniques it may be possible to mitigate the symptoms associated with this destructive virus. Virus resistance may be achieved through the expression of either the sense or antisense viral coat protein gene in orchid tissues if an efficient means of orchid transformation is developed. In this research two transformation protocols were examined for their ability to efficiently transform orchid tissue. The first transformation protocol explored utilized the native ability of Aq bacterium tumefaciens to incorporate DNA into host plants to achieve transformation. The second mechanism explored was particle bombardment transformation.Many strains of A. tumefaciens were employed using direct exposure of Cattleya_ orchid protocorm and callus tissue. Particle bombardment using DNA coated 0.5 um diameter tungsten particles and high pressure helium tank acceleration was employed. The particle bombardment procedure employed the pG35barB plasmid which confers herbicide resistance to the herbicide basta when integrated and expressed in plant tissues.GUS fluorescence assays and PCR analysis indicate that T-DNA is present in orchid tissues, while Southern blot analysis was unable to display that integration had occurred. Particle bombardment yielded herbicide resistant orchid tissues which have yet to be analyzed by Southern blot analysis to confirm integration due to limited tissue quantities. / Department of Biology