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Disease problems in crop plants are still a major threat to the agro-industry worldwide. Various strategies have been developed and evaluated in recent years. One strategy is to use naturally-occurring antipathogen factors such as lysozymes and chitinases in transgenic plants. In the present study, transgenic plants containing chick-egg white lysozyme (Lys 75) have been evaluated for lysozyme production in planta, subcellular localisation, and resistance to some potato pathogens, including Phytophthora infestans and Erwinia carotovora subsp. atroseptica, the two major potato pathogens worldwide. In addition, the evaluation of resistance was also undertaken for transgenic plants carrying other naturally-occurring antipathogen factors including a bean chitinase gene (BCH 35) and a snowdrop lectin gene (GNA 74). In order to accurately quantify the lysozyme production in Lys 75 plants, the turbidimetric lysozyme enzyme assay was optimised. Also, a modified substrate for the enzyme has been developed by covalently linked the Micrococcus lysodeikticus cell wall with a dye, remazol brilliant violet 5R to enable a colorimetric assay of the enzyme. In order to quantitatively assess resistance levels of the transgenic plant, a new method (leaf-bridge bioassay) for conducting and evaluating resistance in planta has been developed. All transgenic plants in tissue culture were tested for resistance using this technique. Evaluation of the progress of infection in detached leaves of Lys 75 showed that lysozyme gave some degree of protection against the bacterial pathogen, Erwinia carotovora subsp. atroseptica and the fungal pathogen, Fusarium sulphureum. Analysis of intercellular fluid from the Lys 75 leaves showed that more than 80% of the total lysozyme expressed in the leaf was located in the intercellular space which is a strategic place to combat pathogen attack. In contrast, the levels of protection in BCH 35 plants were relatively low compared with Lys 75. The progress of infection was delayed in BCH 35 leaves challenged with F. sulphureum only. No resistance at all was observed in GNA 74 to all the pathogens used. All the transgenic potato lines were susceptible to P. infestans. Recently, a new strategy to combat disease problems has been suggested based on a 'durable resistance'. Potato variety Stirling' which shows durable resistance in the field has been used to study the early biochemical and molecular events during elicitation of 'Stirling' cell suspension cultures with an elicitor mix derived from infective units of a compatible strain of P. infestans. For comparison, an elicitor mix from an incompatible strain of P. infestans was also prepared and used. The mixed elicitor comprising zoospore extract, culture filtrate and mycelium homogenate induced defence responses in 'Stirling' cell suspension cultures as judged by the increase in PAL enzyme activity. PAL activity in 'Stirling' ceUs elicited with an elicitor mix derived from an incompatible strain of P. infestans was twice the activity in the compatible interaction. The peak levels in both types of interaction were at 6 h post-elicitation. An oxidative burst was demonstrated also in both types of interactions indicated by rapid release of H(_2)O(_2) into the culture medium. The H(_2)O(_2) level peaked at 2 h post-elicitation in both interactions before being reduced to its normal level at 4 h. The H(_2)O(_2) released during incompatible interaction was twice the levels monitored in the compatible. A subtracted cDNA library of differentially expressed mRNAs during elicitation of 'Stirling' cell suspension cultures with the elicitor mix from a compatible strain of P. infestans was constructed using suppression subtractive hybridisation. Two cDNA clones, STS 42 and STS 52, relevant to the present study were identified and characterised. STS 42 showed high degree of similarity to potato leucine aminopeptidase gene which is induced in response to wounding. Gene expression studies using RT-PCR showed that the mRNA levels of STS 42 increased gradually throughout the 18 h elicitation. STS 51 was identified as a member of the ribonuclease T2 histidine proteins. It showed some degree of similarity to plant ribonucleases involved in self-incompatibility reactions during pollination. It has a site for tyrosine kinase phosphorylation at the hydrophilic region of the sequence and could possibly be involved in phosphorylation during signal transduction. mRNA levels of STS 51 were increased during the first 12 h of elicitation.

published_or_final_version / Ecology and Biodiversity / Doctoral / Doctor of Philosophy

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Thesis (Ph. D.)--University of Hong Kong, 2002. / Includes bibliographical references (leaves 91-107).

Thesis (Ph.D.)--University of Hong Kong, 2002. / Includes bibliographical references (leaves 91-107) Also available in print.

Thesis (Ph. D.)--Ohio State University, 2005. / Document formatted into pages; contains 215 p. Includes bibliographical references. Abstract available online via OhioLINK's ETD Center; full text release delayed at author's request until 2006 March 3.

No description available.

Doctor of Philosophy / Department of Plant Pathology / Karen A. Garrett / Late blight of potato, caused by Phytophthora infestans, is a pernicious disease of potatoes worldwide. This disease causes yield losses as a result of foliar and tuber damage. Many models exist to predict late blight risk for control purposes with-in season but rely upon fine-scale weather data collected in hourly, or finer, increments. This is a major constraint when working with disease prediction models for areas of the world where hourly weather data is not available or is unreliable. Weather or climate summary datasets are often available as monthly summaries. These provide a partial solution to this problem with global data at large time-steps (e.g., monthly). Difficulties arise when attempting to use these forms of data in small temporal scale models. My first objective was to develop new approaches for application of disease forecast models to coarser resolution weather data sets. I created metamodels based on daily and monthly weather values which adapt an existing potato late blight model for use with these coarser forms of data using generalized additive models. The daily and monthly weather metamodels have R-squared values of 0.62 and 0.78 respectively. These new models were used to map global late blight risk under current and climate change scenarios resistant and susceptible varieties. Changes in global disease risk for locations where wild potato species are indigenous, and disease risk for countries where chronic malnutrition is a problem were evaluated. Under the climate change scenario selected for use, A1B, future global late blight severity decreases. The risk patterns do not show major changes, areas of high risk remain high relative to areas of low risk with rather slight increases or decreases relative to previous years. Areas of higher wild potato species richness experience slightly increased blight risk, while areas of lower species richness experience a slight decline in risk.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. Programa de Pós-graduação em Engenharia de Produção / Made available in DSpace on 2012-10-20T21:06:49Z (GMT). No. of bitstreams: 1 225932.pdf: 1800466 bytes, checksum: 7546469656bbd6296a6ac29ff925f29e (MD5) / Este trabalho apresenta um estudo desenvolvido com dados longitudinais, usando a metodologia de modelos não lineares em curvas de crescimento, com diferentes estruturas para a matriz de covariância, fixando uma função para a parte determinística. Após a seleção da melhor matriz de covariância, foram experimentadas diferentes funções, a fim de se escolher o modelo não linear mais adequado. Este procedimento foi aplicado em dados da porcentagem de severidade da doença Late blight em quatro variedades de batata (Solanum tuberosum), causada por Phytophthora infestans. O modelo não linear selecionado foi o que usa uma das parametrizações da função de Gompertz e matriz de covariância de Simetria Composta. Os testes de hipóteses realizados sobre os parâmetros do modelo confirmaram a existência de diferença significativa entre as variedades.