DEFENCE GENE EXPRESSION IN THE TOMATO-VERTICILLIUM PATHOSYSTEM

The Atrium, University of Guelph Institutional Repository

DEFENCE GENE EXPRESSION IN THE TOMATO-VERTICILLIUM PATHOSYSTEM

Show simple item record

dc.contributor.advisor Robb, Jane
dc.contributor.author Castroverde, Christian Danve
dc.date 2010-03-15
dc.date.accessioned 2010-04-22T19:25:45Z
dc.date.available 2010-04-22T19:25:45Z
dc.date.issued 2010-04-22T19:25:45Z
dc.identifier.uri http://hdl.handle.net/10214/2146
dc.description.abstract In tomato (Solanum lycopersicum), race-specific resistance against the fungal wilt pathogen Verticillium dahliae race 1 (Vd1) is established in the stem. However, the molecular factors and mechanisms leading to this resistance response are still unknown. In this study, Craigella resistant (CR) and susceptible (CS) tomato plants were successfully infected with Vd1 and this was verified by fungal quantification and symptom score assays. Previous microarray results showed interesting patterns of defence gene expression that correlated with biological phenomena. Plant defence genes code for proteins that are responsible for or associated with the plant resistance response. Through RT-PCR, this thesis set out to confirm these microarray observations and also to generate expression data for genes in which sensitivity was an issue in the microarray. The standard RT-PCR data confirmed a number of the microarray results, but some conflicts remained. From the defence genes investigated, there was agreement between the microarray data and the RT-PCR data for pre-mRNA processing factor 8, class IV chitinase, cyclin-dependent kinase inhibitor and IMP dehydrogenase/GMP reductase. Partial agreement was observed for genes coding for ethylene response factor 2, phenylalanine ammonia lyase and P6 protein. However, there was total disagreement for 14-3-3, beta-glucanase, P1a, RNA-binding protein, calcium-binding protein and S-Adenosyl-L-methionine: hydroxide adenosyltransferase. Real-time RT-PCR was attempted to clarify the remaining issues but further discrepancies arose, particularly in the Ve resistance genes. To resolve these discrepancies, two approaches were designed: (1) one based on the use of a universal internal control and (2) another based on restriction enzyme digestion. In general, the results were more consistent with standard RT-PCR. Overall, this study showed that standardization of a system involving vascular pathogens, leading to reproducible analysis, was possible but only with proper controls and additional validation. Standard RT-PCR appeared to offer a more accurate picture of the expression of defence genes in the tomato-Verticillium pathosystem. The defence gene expression results confirmed in this study remain as potential insights into the molecular mechanisms for Verticillium resistance in tomato plants. en
dc.language.iso en en
dc.rights.uri http://creativecommons.org/licenses/by-nc-nd/3.0/ *
dc.subject Real-time PCR en
dc.subject Microarray en
dc.subject RFLP en
dc.subject Internal control en
dc.subject Plant resistance en
dc.subject RT-PCR en
dc.subject Ve genes en
dc.subject Gene expression en
dc.subject Defence genes en
dc.subject Plant defence en
dc.subject Solanum lycopersicum en
dc.subject Verticillium en
dc.title DEFENCE GENE EXPRESSION IN THE TOMATO-VERTICILLIUM PATHOSYSTEM en
dc.type Thesis en
dc.degree.programme Molecular and Cellular Biology en
dc.degree.name Master of Science en
dc.degree.department Department of Molecular and Cellular Biology en


Files in this item

Files Size Format View Description
THESIS final.pdf 2.483Mb PDF View/Open Master's Thesis - Castroverde

This item appears in the following Collection(s)

Show simple item record

http://creativecommons.org/licenses/by-nc-nd/3.0/ Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-nd/3.0/

Search the Atrium


Advanced Search

Browse

My Account