[No abstract available] / xi, 176 leaves : ill. (chiefly col.) ; 29 cm

Turnip crinkle virus (TCV) has been implicated in the suppression of the hypersensitive response (HR), a type of programmed cell death induced during active resistance in Arabidopsis thaliana. In order to investigate the involvement of individual viral components in mediating suppression, TCV genes were cloned for use in an Agrobacterium tumefaciens mediated transient expression in Nicotiana benthamiana. Agroinfiltration of the HR-inducing avrPto/Pto system in conjunction with individual TCV genes has identified the p38 gene, which encodes the viral coat protein, as the gene responsible for the cell death suppression phenotype. The extent of cell death suppression by coat protein was quantified and found to be equal to the level of suppression by the whole virus and AvrPtoB, another cell death inhibitor from bacteria. Thus, the coat protein alone is sufficient to inhibit the HR in plants. Further, the effect of TCV on HR initiation by an avirulence factor from an unrelated bacterial pathogen was investigated. The presence of TCV does not affect the production, secretion or cellular processing of the bacterial avirulence factor.

Shuk-man Chow. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 171-186). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgement --- p.viii / General abbreviations --- p.x / Abbreviations of chemicals --- p.xi / List of figures --- p.xii / List of Tables --- p.xiii / Table of contents --- p.xv / Chapter 1. --- Literature review / Chapter 1.1. --- General introduction to rice disease --- p.1 / Chapter 1.1.1. --- Pathogenesis of Bacterial Leaf Blight (BLB) --- p.1 / Chapter 1.1.2. --- Pathogenesis of rice blast --- p.2 / Chapter 1.1.3. --- Control of rice diseases --- p.3 / Chapter 1.2. --- Plant defense mechanisms --- p.4 / Chapter 1.2.1. --- Basal resistance in plants --- p.4 / Chapter 1.2.2. --- Wound induced defense response --- p.5 / Chapter 1.2.3. --- Pathogen induced host defense response --- p.6 / Chapter 1.3. --- Structure of R gene products --- p.7 / Chapter 1.4. --- Recognition between R and Avr proteins in rice --- p.8 / Chapter 1.5 --- Current knowledge on Xa resistance and AvrXa avirulence protein --- p.9 / Chapter 1.6 --- Current knowledge on Pi resistance and AvrPi avirulence protein --- p.10 / Chapter 1.7 --- Pathogen induced signal transduction cascade --- p.12 / Chapter 1.7.1. --- R gene mediated signal transduction cascade --- p.12 / Chapter 1.7.2. --- Signal events of G-protein activation --- p.12 / Chapter 1.7.3. --- Signaling events for the accumulation of Ca2+ in cytosol --- p.13 / Chapter 1.7.4. --- Signaling events for oxidative burst --- p.14 / Chapter 1.7.5. --- MAPK cascade in defense signaling --- p.15 / Chapter 1.7.6. --- Transcriptional regulation of disease resistance related genes --- p.16 / Chapter 1.7.7. --- Translational regulation of disease resistance related genes --- p.17 / Chapter 1.8. --- Defense responses and defense related genes --- p.19 / Chapter 1.8.1. --- Pathogenesis related (PR) proteins --- p.20 / Chapter 1.8.2. --- Phytoalexins --- p.21 / Chapter 1.9. --- Disease resistance related genes common between rice blast and BLB resistance --- p.22 / Chapter 1.10. --- SA induced signal transduction pathway in rice --- p.23 / Chapter 1.11. --- Important tools facilitating the identification of disease resistance related genes from BLB resistant rice lines --- p.24 / Chapter 1.12. --- Hypothesis --- p.26 / Chapter 1.13. --- Project objective --- p.26 / Chapter 2. --- Materials and Methods --- p.27 / Chapter 2.1. --- Plant Materials --- p.27 / Chapter 2.2. --- Pathogen Inoculation --- p.27 / Chapter 2.3. --- RNA extraction --- p.29 / Chapter 2.4. --- Denaturing gel electrophoresis --- p.29 / Chapter 2.5. --- Subtraction libraries construction --- p.30 / Chapter 2.5.1. --- Cloning of disease resistance related genes --- p.32 / Chapter 2.5.1.1. --- pBluescript II KS (+) T-vector preparation --- p.32 / Chapter 2.5.1.2. --- Ligation --- p.32 / Chapter 2.5.1.3. --- Transformation --- p.32 / Chapter 2.5.1.4. --- Colony picking --- p.33 / Chapter 2.5.1.5. --- PCR amplification of DNA inserts --- p.33 / Chapter 2.5.1.6. --- Purification of PCR products --- p.34 / Chapter 2.6. --- Gene chips printing --- p.34 / Chapter 2.7. --- Probes synthesis and gene chips hybridization --- p.35 / Chapter 2.8. --- Standard-RNAs synthesis --- p.35 / Chapter 2.9. --- Data collection and analysis --- p.36 / Chapter 2.10. --- Sequencing --- p.36 / Chapter 2.11. --- cDNA synthesis --- p.37 / Chapter 2.12. --- RT-PCR --- p.38 / Chapter 2.13. --- DNA gel electrophoresis --- p.39 / Chapter 3. --- Results --- p.58 / Chapter 3.1. --- Construction of BLB gene chips --- p.58 / Chapter 3.1.1. --- Preparation of cDNA clones for gene chips construction --- p.58 / Chapter 3.1.2. --- Purification of PCR products on microtiter plate --- p.59 / Chapter 3.1.3. --- Gene chips construction --- p.59 / Chapter 3.1.4. --- DNA immobilization --- p.62 / Chapter 3.1.5. --- Probe synthesis --- p.62 / Chapter 3.1.6. --- Gene chip analysis --- p.65 / Chapter 3.1.6.1. --- Scanning --- p.65 / Chapter 3.1.6.2. --- Data analysis --- p.65 / Chapter 3.2. --- "Identification of disease resistance related genes commonly regulated by Xa2, Xal2 and Xal4 BLB resistance loci" --- p.70 / Chapter 3.2.1. --- "Signal perception, transduction and regulatory elements" --- p.71 / Chapter 3.2.1.1. --- Proteins involved in reversible phosphorylation cascade --- p.71 / Chapter 3.2.1.2. --- Proteins potentiate signal transduction through specific protein-protein interaction --- p.72 / Chapter 3.2.1.3. --- Other signal transduction components --- p.73 / Chapter 3.2.2. --- Transcriptional and translational regulatory elements --- p.74 / Chapter 3.2.2.1. --- Proteins involved in transcriptional regulation --- p.74 / Chapter 3.2.2.2. --- Proteins involved in post-transcriptional regulation --- p.75 / Chapter 3.2.2.3. --- Proteins involved in translational regulation --- p.76 / Chapter 3.2.3. --- "Oxidative burst, stress, apoptotic related genes" --- p.77 / Chapter 3.2.3.1. --- Stress related proteins --- p.77 / Chapter 3.2.3.2. --- Proteins involved in induction of oxidative burst --- p.78 / Chapter 3.2.3.3. --- PR proteins --- p.79 / Chapter 3.2.3.4. --- Proteolysis related proteins --- p.79 / Chapter 3.2.4. --- Cell maintenance and metabolic genes --- p.80 / Chapter 3.2.4.1. --- Antioxidant --- p.80 / Chapter 3.2.4.2. --- Metabolic genes --- p.81 / Chapter 3.2.4.3. --- Molecular chaperone --- p.82 / Chapter 3.2.4.4. --- Cell cycle regulators --- p.82 / Chapter 3.2.4.5. --- Cell wall maintenance --- p.83 / Chapter 3.2.4.6. --- Proteins involved in protein transport --- p.83 / Chapter 3.2.5. --- Unclassified/others --- p.84 / Chapter 3.3. --- Expression analysis of disease resistance related genes --- p.88 / Chapter 4. --- Discussion --- p.141 / Chapter 4.1. --- Differential expression of disease resistance candidates --- p.141 / Chapter 4.2. --- Disease resistance signal transduction components --- p.143 / Chapter 4.2.1. --- Reversible phosphorylation cascade --- p.143 / Chapter 4.2.2. --- Signal transduction potentiated by protein-protein interaction --- p.144 / Chapter 4.3. --- Other signaling molecules --- p.145 / Chapter 4.3.1. --- PRL1-interacting factor G --- p.145 / Chapter 4.3.2. --- Vacuolar-type H+-ATPasen subunit G --- p.146 / Chapter 4.4. --- Regulation of expression of disease resistance candidates --- p.146 / Chapter 4.4.1. --- Transcriptional regulation of disease resistance related genes --- p.146 / Chapter 4.4.1.1. --- G-box binding protein --- p.147 / Chapter 4.4.1.2. --- MYB TF --- p.147 / Chapter 4.4.2. --- Post-transcriptional modification of disease resistance candidates --- p.148 / Chapter 4.4.2.1. --- RNA splicing factor --- p.148 / Chapter 4.4.2.2. --- Glycine rich RNA binding proteins --- p.149 / Chapter 4.4.3. --- Translational regulation of disease resistance related genes --- p.149 / Chapter 4.5. --- Induction of oxidative burst --- p.150 / Chapter 4.6. --- PR proteins --- p.151 / Chapter 4.7. --- Cell maintenance --- p.152 / Chapter 4.7.1. --- Protein folding --- p.152 / Chapter 4.7.2. --- Protein degradation --- p.153 / Chapter 4.7.3. --- ROS scavenging --- p.154 / Chapter 4.7.4. --- Regulation of cell cycle --- p.154 / Chapter 4.8. --- "Confirmation and profiling of disease resistance related candidates commonly regulated in Xa2, Xal2 and Xal4 BLB resistance NILs at different time points" --- p.155 / Chapter 4.8.1. --- Basal resistance related genes --- p.156 / Chapter 4.8.2. --- General disease resistance related genes --- p.161 / Chapter 4.8.3. --- Pathogen responsive genes --- p.164 / Chapter 4.8.4. --- Prediction of novel genes functions --- p.168 / Chapter 4.9. --- Future prospect --- p.169 / Chapter 4.10. --- Conclusion --- p.169 / References --- p.171 / Appendix --- p.187

No description available.

No description available.

Graduation date: 2003

No description available.

An interaction was recently detected between the potyviral protein, genome-linked (VPg) and the Arabidopsis thaliana translation initiation factor eIF(iso)4E (Wittmann et al., 1997). / Here, experiments were undertaken to address biological aspects of the VPg-eIF4E interaction. First, coimmunoprecipitation experiments performed with purified recombinant proteins have shown that VPg not only associates with eIF4E, as was previously published, but also with the larger eIF4F complex, of which eIF4E is a subunit. These results were confirmed by ELISA-type binding assays. It was also shown that there is no direct interaction between VPg and the other subunit of eIF4F, namely eIF4G. Finally, with the same experimental system, it was shown that the presence of eIF4G does not influence the binding affinity of VPg and eIF4E. / The interaction of VPg with the plant translation apparatus suggests that potyviral infection may alter the host protein expression profile. This hypothesis was investigated with the use of a protoplast system. We have shown that the global rates of protein synthesis in protoplasts transfected with an infectious TuMV cDNA clone dropped shortly after transfection, by as much as an estimated 70%. Recovery to normal levels occurred within 48 hours. / Evidence was obtained that the interaction between VPg and eIF4E is instrumental in this transient down-regulation of protein expression: protoplasts transfected with a mutant TuMV cDNA clone, the VPg of which has no affinity for eIF4E, failed to exhibit the drop in protein synthesis observed with the wild-type clone.

Turnip mosaic potyvirus (TuMV) is a single-stranded RNA plant virus. One of its proteins, the cylindrical inclusion (CI) protein, was hypothesized to interfere with host transcription via interaction with histone H3 protein. Interaction between CI and histone H3 was previously observed in Dr. Fortin's laboratory. Based on previous studies that demonstrated the importance of the H3 tail domain in gene regulation and chromosome arrangement, it was hypothesized that CI would interact with the tail rather than the globular domain. The objective of this project was to identify which histone H3 domains CI protein interacts with. The full-length, globular, and tail domains of histone H3 DNA were expressed in E. coli and purified. Based on in vitro interaction experiments, the CI protein was observed to interact with the globular domain of histone H3.

Turnip mosaic virus (TuMV) infects a variety of crops, worldwide, including the economically relevant Brassicacea family. It was previously demonstrated that TuMV infection in tobacco protoplasts leads to an overall decrease of host protein. However, it remains unclear whether this phenomenon is due to the repression of plant gene transcription during the infection period or due to viral inhibition of host translation. In this study, quantification of various transcripts and protein products from infected tobacco was performed via real-time RT-PCR and ELISA. In comparison to the gamma-tubulin endogenous control, gene expression for the tobacco H3, HSP70 and granule-bound starch synthase was affected by TuMV infection with time. / Tobacco protein accumulation in whole leaf tissues was also significantly affected by increase of virus particles.