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Title: Development of diagnostic techniques for Pyrenopeziza brassicae on winter oilseed rape
Author: Foster, S. J.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2000
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Using sequence information from the mating type idiomorphs (MAT-1 and MAT-2) of P. brassicae, the causal agent of light leaf spot of winter oilseed rape (Brassica napus ssp. oleifera), two PCR-based diagnostic assays were developed. The first assay was designed as a general diagnostic tool for P. brassicae and was shown to be specific, amplifying a 750 base pair (bp) product from genomic DNA from over 50 isolates of P. brassicae and not from DNA from a number of other fungal species pathogenic on oilseed rape or from the closely related discomycete Tapesia yallundae. The second assay was designed to differentiate between the two mating types (MAT-1 and MAT-2) of P. brassicae, and was also specific to P. brassicae. Both assays were ale to detect the presence of P. brassicae from DNA extracted from oilseed rape leaves with light leaf spot symptoms. The sensitivities of both of the assays were increased by using templates consisting of mixtures of NDA from P. brassicae and B. napus compared with DNA from P. brassicae alone. Using a nested PCR approach, the sensitivity of the general diagnostic assay was increased by two orders of magnitude and it was able to detect the presence of P. brassicae prior to the development of visible symptoms on oilseed rape leaves. As a first step towards the development of an immuno-diagnostic tool for light leaf spot, the feasibility of adopting a targeted approach, combining molecular and immunological techniques, was investigated.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available