Call number: LD2668 .T4 1952 H34 / Master of Science

Broccoli head rot is a destructive disease found in most broccoli production areas. The main pathogen is the bacterium Pseudomonas fluorescens. P. fluorescens 5064, which was first isolated from an infected broccoli head in SE Scotland, produces biosurfactants that are important for bacterial establishment on the plant surface prior to causing disease in broccoli. Preliminary experiments performed in this study showed that biosurfactant production in P. fluorescens 5064 was cell density dependent, which is a typical characteristic of the quorum sensing mechanism. Quorum sensing is a bacterial communication mechanism, which controls a number of key processes in growth, reproduction and virulence via signalling molecules (quorum sensing signal) in many gram-negative bacteria. One aim of this study was to determine if biosurfactant production in P. fluorescens 5064 is controlled via quorum sensing. To do this, 35 surfactant-minus Tn5 mutants of P. fluorescens 5064 were screened for their abilities to produce a quorum sensing signal. Six of these biosurfactant-deficient mutants showed a large reduction in quorum sensing signal production. In one mutant 6423, which contains a single Tn5 insertion, the production of the quorum sensing signal was almost eliminated. Addition of quorum sensing signal, either synthetic or extracted from wild type P. fluorescens 5064, was able to restore biosurfactant production in mutant 6423. This strongly suggests that quorum sensing regulates biosurfactant production in P. fluorescens 5064. Attempts were made to clone and sequence the Tn5 disrupted gene in mutant 6423, but the identity of the gene remains inconclusive. The quorum sensing signal in wild type P. fluorescens 5064 was identified in this study by High Pressure Liquid Chromatography and Mass Spectrometry as N-3-hydroxyoctanoyl-homoserine lactone, which has been shown by other researchers to be present in P. fluorescens strain 2-79, but not in the strains F113, 7-14 and NCIMB 10586. The discovery that biosurfactant production in P. fluorescens 5064 is regulated by quorum sensing opens up a possibility for novel control of broccoli head rot. Although only the control of biosurfactant production by quorum sensing was examined in this study, it is possible that other virulence factors, such as pectic enzyme production, are also controlled by quorum sensing as in other pathogenic bacteria. By blocking the quorum sensing system, the pathogenic P. fluorescens that use this mechanism to control virulence could potentially be rendered avirulent. In greenhouse pathogenicity tests, a quorum sensing signal-degrading bacterium, Bacillus sp. A24, was evaluated for biocontrol of head rot disease caused by P. fluorescens 5064 on broccoli. However, the Bacillus sp. A24 showed only limited control effects, despite its strong quorum sensing signal-degrading ability towards the pathogen in vitro. A subsequent test proved that Bacillus sp. A24 is a surfactant producer itself and this could explain its ineffectiveness in disease control. When screening the quorum sensing signals of the 35 biosurfactant mutants, mutant 6418 was found to produce a potent antibiotic-like compound. This was identified by thin-layer chromatography as pyrrolnitrin. Unlike wild-type P. fluorescens 5064, mutant 6418 has lost its ability to produce virulence factors and is thus non-pathogenic. It was therefore of interest to determine if mutant 6418 could be used as a biocontrol agent to control broccoli head rot disease. In greenhouse pathogenicity tests, mutant 6418 significantly reduced disease by 41 %. The practical application of this research to bacterial disease control – via the manipulation of quorum sensing to inhibit virulence gene expression – is discussed.

Vita. / Digitized by Kansas Correctional Industries

Background: Perinatal ischemic brain injury leads to developmental disability (DD), which accounts for 30% of disabilities in children. Antepartum risk, or risk occurring prior to birth occurs in more than 90% of cases. This study investigated whether maternal ingestion of a natural health product (broccoli sprouts) would provide neuroprotection in an intrauterine model of HI. Methods: Intrauterine ischemia was induced by bilateral uterine artery ligation (BUAL) on E20 of gestation. Rats were fed broccoli sprouts (200 mg) from E15 until postnatal day 14 (PD14). Rat pups underwent neurobehavioural testing from birth to PD21 and were then sacrificed for neuropathologic assessment on PD21. Results: BUAL ligation resulted in growth restriction (IUGR) of the fetuses, which persisted throughout the study (p < 0.001). Reflex testing indicated IUGR pups were developmentally delayed compared to controls (p < 0.001). Open field testing on PD21 indicated hyperactivity in IUGR animals compared to controls (p < 0.001). Histological assessment showed a reduction in pyramidal cells in CA1 and CA3 of IUGR hippocampi and in myelin basic protein (MBP) immunohistochemistry signal. Broccoli sprout supplementation improved some reflex and behavioural measures, increased cell counts in CA1 and CA3 as well as MBP signal in growth restricted animals. Conclusions: Supplementation with broccoli sprouts during the last trimester of gestation and the first 2 weeks of life in the rat lessened the effects of chronic intra-uterine ischemia. These findings suggest a novel approach to the prevention of DD associated with perinatal HI.

Organic agriculture is an important and growing sector of U.S. and world food production. Consumers are increasingly aware of and interested in the production practices and impacts associated with agriculture and as such, are showing a preference for sustainably produced, raised, and harvested foods. In order to continue to meet the growing demand for organic produce, organic growers need cultivars that are optimally adapted to organic and low input conditions. Quality seed is the foundation of any functional and stable farming system. Unfortunately the lack of organically bred and produced seed is hindering the continued growth and success of organic farming. Meeting the needs of the organic sector has been a challenge for the seed industry; it is an industry that often doesn't understand the specific and unique requirements associated with the diversity of environmental and market demands of organic systems. However, organic farmers and the organic food systems they supply, require a robust organic seed system that is appropriately adapted to regional agronomic challenges and market needs, meets standards and regulations, and encompasses the social and ecological values of organic agriculture. One plausible approach to meeting the cultivar and seed needs of organic and low input production systems is through the use of participatory plant breeding (PPB). PPB is a collaborative approach for identifying and developing genetically diverse plant material and varieties involving partnerships among formal sector breeders and researchers, farmers, extension agents, educators, and end users. Participatory plant breeding fundamentally changes the way that formal breeding programs and farmers manage germplasm and plant genetic resources. Typically, formal breeding programs restrict access to germplasm and breeding materials and only supply farmers with finished varieties. In PPB, farmers are involved in the early stages of creation and evaluation of germplasm and breeding material, and stay engaged with the breeding process until new varieties are created. PPB is an excellent model for breeding specifically for organic systems because organic systems in developed countries have many similarities to low-input agricultural systems in the developing world. Some of these parallels include heterogeneous growing environments, a wide range of end uses and marketing strategies, lack of suitably adapted and/or derived varieties, lack of attention from the formal seed sector, and a reduced reliance on synthetic inputs (compared to conventional systems). Breeding for organic systems is a relatively young field and breeders in the formal sector do not have a good handle on what traits are important for robust production under organic conditions. Thus the opportunity to meld farmers' experience and knowledge with breeders' expertise is an effective way to breed for organic production systems. The purpose of this project was to investigate and explore the opportunities and challenges of organic plant breeding using participatory research methods. This research had three goals: 1) to develop an open pollinated broccoli with contemporary quality traits for organic production systems using participatory strategies; 2) to compare broccoli selections made by formally trained plant breeders and farmer breeders; and 3) to capture the stories and experiences of the formal breeders and farmer breeders involved with this broccoli material in order to contribute to the growing wealth of knowledge on collaborative and organic breeding work. The Oregon State University Vegetable Breeding Program made significant progress towards decreasing the variability of the broccoli project material through three successive years of modified half-sibling selections. Evaluations and selections were based more strongly on quality traits rather than soley on production traits such as yield. Although progress was incremental and statistically verified in only three out of the fifteen quality traits, we observed trends in the data indicating progress towards an increasingly uniform, stable, and reliable open pollinated broccoli with specific adaptation for organic production systems. There were very few differences between broccoli materials developed by formally trained plant breeders and farmer breeders. This was especially true for the three cultivars developed in the Pacific Northwest (PNW) (one in Washington and two in Oregon). The 'East Coast' population, which had been collaboratively selected by formal and farmer breeders in New York, expressed significantly distinct differences from the PNW materials. When the farmer breeder and formal breeder materials were pooled together and compared to pooled check cultivars they expressed significant differences for nearly all traits across all years. This demonstrated that all of the collaboratively developed open pollinated materials are distinctly different from the F1 hybrids currently available. Our work has demonstrated a few of the myriad of positive outcomes achievable with the use of participatory plant breeding for organic production systems. The participatory nature of this project resulted in increased confidence and feelings of empowerment for all involved. Both farmers and breeders felt their involvement was socially beneficial and widened their networking and seed community circles. The farmer-bred cultivar 'Solstice' is now available as a result of Jonathan Spero's work, and a cultivar tentatively named 'Benton' is about to be released for sale through Oregon State University. Our results agree with previous study findings that formal and farmer breeder selections are often not distinctly different; thus providing evidence for continuing to support the involvement and education of farmers in plant breeding, especially in reference to organic production systems. This study demonstrates the potential of collaboratively developed and farmer-bred cultivars to become viable and vibrant open pollinated alternatives to the current open pollinated cultivars on the market today. / Graduation date: 2013

This item was digitized as part of the Million Books Project led by Carnegie Mellon University and supported by grants from the National Science Foundation (NSF). Cornell University coordinated the participation of land-grant and agricultural libraries in providing historical agricultural information for the digitization project; the University of Arizona Libraries, the College of Agriculture and Life Sciences, and the Office of Arid Lands Studies collaborated in the selection and provision of material for the digitization project.

No description available.

Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1992. / Vita. Abstract. Includes bibliographical references (leaves 115-117). Also available via the Internet.

The first aim of this study was to determine the effect of irrigation intervals on the survival of L. monocytogenes on spray irrigated broccoli under field trial conditions, and subsequent survival of the pathogen on broccoli during postharvest processing procedures. The nonpathogenic L. innocua was used as surrogate organism to L. monocytogenes. Broccoli in the field was treated with irrigation water inoculated with L. innocua, during intervals over a period of five weeks and the growth and survival of the organism was monitored weekly. L. innocua numbers remained similar over intervals that received consecutive inoculations and L. innocua numbers decreased by at least 2.3 log cfu/g after inoculation ceased, which showed an inoculation effect and that time had an influence on organism survival. Cessation of irrigation before harvest was found to effectively reduce pathogen contamination levels on the crop, whilst repeated irrigation with contaminated water contributes to maintenance of L. innocua as well as elevated total microbial counts on the broccoli. A lack of correlation between the L. innocua counts and the recorded environmental temperatures in the field, including temperature and relative humidity, suggested that survival is not solely dependent on and influenced by, nor can it be predicted by these parameters. It was found that the presence of high levels of contamination (with, in this case L. innocua) in irrigation water used for vegetable crops, can be associated with an increased microbial population on the crop surface. Secondly, the effect of processing on organism survival post-harvest was assessed. Washing with water caused a 1 log reduction of L. innocua, whilst washing with 200 ppm chlorinated water facilitated a further 1 log reduction. Cooking reduced L. innocua numbers on broccoli by an average of 1.1 log units and aerobic plate counts by between 1 and 2 log units. A combined treatment of washing with chlorine, storage in MAP (5% CO2, 5% O2) for two days at 4°C and final microwave heating resulted in the lowest pathogen numbers, causing a 5.13 log cfu/g log reduction. Therefore, even though chlorine isprocessing, it does not suffice alone to eliminate pathogens (with L. innocua being representative of L. monocytogenes) from vegetables, just as MAP storage is only effective as part of a hurdle procedure. Cooking is essential in destroying L. innocua present on broccoli and to ensure vegetables that are safe for consumption in terms of pathogenic exposure. With this knowledge on the behaviour of L. monocytogenes on broccoli, the risk associated with the application of contaminated irrigation water to fresh produce can be better understood and the hazard managed. Copyright / Dissertation (MSc)--University of Pretoria, 2012. / Food Science / unrestricted

Broccoli (<i>Brassica oleracea</i> L. var. italica) has become an increasingly popular vegetable with American consumers. Much of the attractiveness of fresh broccoli is derived from this vegetable's high nutrition and excellent organoleptic properties. In a consumer response survey, Virginia Master Gardeners indicated a preference for broccoli that has less stalk and more florets by weight. The objectives of this research were to produce single unit broccoli, and to examine vacuum and shrink-film wrapping of single unit and bunching broccoli as alternatives to the standard post harvest practice of top icing in order to preserve post harvest quality attributes: vitamin C, color, odor, and turgidity. Broccoli cv. 'Symphony' was direct seeded and transplanted at two sites in Virginia (within row spacing 20 cm , .9 m centered, 3 row bed). Single unit broccoli was classified as broccoli with head diameters greater than or equal to 20 cm (i.e., equivalent to one bunch of broccoli). Single unit and bunching broccoli for post harvest experimentation was vacuum or shrink-film wrapped with plastic film wraps of varying gauges. The broccoli was held in cold storage from 12 to 22 days with no supplemental ice. Single unit broccoli was produced successfully at one site only. The yields, however, were a fraction of total bunching broccoli yields. There was no significant difference in marketable yields with either planting method (direct seeded vs transplanting) at site 2. Transplants, however, out yielded the direct seeded broccoli at site 1. Vacuum and shrink-film wrapping proved to be very effective in preserving organoleptic attributes of single unit and bunching broccoli even when never receiving ice. Vitamin C and chlorophyll retention were not influenced by wrapping. / Master of Science