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Comparison of molecular techniques for the identification of DNA markers specific to Fusarium oxysporum f. sp. cyclaminis Louie, Daryl Andy

Abstract

Fusarium wilt of cyclamen is caused by the fungus Fusarium oxysporum f. sp. cyclaminis. Early detection of this disease in greenhouse plantings has been difficult due to the long latent period which may occur and the ubiquity of morphologically identical nonhost F. oxysporum strains present in the greenhouse. A PCR based assay to confirm the identity of F. o. cyclaminis would be an improvement over any conventional methods due to its inherent speed, specificity, and sensitivity. The molecular techniques of random amplified polymorphic DNA (RAPD), combined polymerase chain reaction and restriction fragment length polymorphism (PCR and RFLP), and subtraction hybridization were used to attempt identification of DNA markers specific to F. o. cyclaminis. No universal DNA markers were found which would identify the 16 F. o. cyclaminis isolates from the 25 nonhost Fusarium isolates used in this study. The complicating factor appeared to be the apparent loss of pathogenicity with some of the F. o. cyclaminis isolates. Of the methods evaluated, RAPD analysis or PCR and RFLP analysis using the intergenic spacer (IGS) region, proved to be the most promising methods due to their ease of use. Cluster analysis of the RAPD data using the unweighted paired group method with arithmetic averaging (UPGMA) revealed that the pathogenic F. o. cyclaminis isolates were found to be exclusive to two clusters. Future research towards identification of DNA markers to this pathogen may best be approached by separation of the isolates with highest genetic similarity into groups and identifying DNA markers to these groups rather than identifying a universal marker to all F. o. cyclaminis isolates.

Item Metadata

Title
Comparison of molecular techniques for the identification of DNA markers specific to Fusarium oxysporum f. sp. cyclaminis
Creator
Louie, Daryl Andy
Publisher
University of British Columbia
Date Issued
1995
Description
Fusarium wilt of cyclamen is caused by the fungus Fusarium oxysporum f. sp. cyclaminis. Early detection of this disease in greenhouse plantings has been difficult due to the long latent period which may occur and the ubiquity of morphologically identical nonhost F. oxysporum strains present in the greenhouse. A PCR based assay to confirm the identity of F. o. cyclaminis would be an improvement over any conventional methods due to its inherent speed, specificity, and sensitivity. The molecular techniques of random amplified polymorphic DNA (RAPD), combined polymerase chain reaction and restriction fragment length polymorphism (PCR and RFLP), and subtraction hybridization were used to attempt identification of DNA markers specific to F. o. cyclaminis. No universal DNA markers were found which would identify the 16 F. o. cyclaminis isolates from the 25 nonhost Fusarium isolates used in this study. The complicating factor appeared to be the apparent loss of pathogenicity with some of the F. o. cyclaminis isolates. Of the methods evaluated, RAPD analysis or PCR and RFLP analysis using the intergenic spacer (IGS) region, proved to be the most promising methods due to their ease of use. Cluster analysis of the RAPD data using the unweighted paired group method with arithmetic averaging (UPGMA) revealed that the pathogenic F. o. cyclaminis isolates were found to be exclusive to two clusters. Future research towards identification of DNA markers to this pathogen may best be approached by separation of the isolates with highest genetic similarity into groups and identifying DNA markers to these groups rather than identifying a universal marker to all F. o. cyclaminis isolates.
Extent
4670449 bytes
Genre
Thesis/Dissertation
Type
Text
File Format
application/pdf
Language
eng
Date Available
2009-01-27
Provider
Vancouver : University of British Columbia Library
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
DOI
10.14288/1.0086949
URI
http://hdl.handle.net/2429/3914
Degree
Master of Science - MSc
Program
Plant Science
Affiliation
Land and Food Systems, Faculty of
Degree Grantor
University of British Columbia
Graduation Date
1995-11
Campus
UBCV
Scholarly Level
Graduate
Aggregated Source Repository
DSpace

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For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.