Project reference: 2007-04
DNA–based identification of needle pathogens (Massey University)
Project description:
Objective: Improved identification capabilities for fungal needle pathogens:
Species–specific DNA diagnostic assay to detect D. septosporum (previously called D. pini) in infected needles.
Mating–type specific assay to monitor any appearance of MAT1 D. septosporum in NZ.
In the longer term diagnostic assays could be developed for other needle pathogens and quantitative as well as qualitative assays could be performed.
Being PCR–based, these assays will be sensitive and capable of detecting very low levels of infection.
Background: DNA extracted from pine needles is usually of poor quality due to the high resin and polysaccharide content of the needles. A masters student, Naydene Barron, developed a simple but promising technique for extracting DNA from Dothistroma–infected needles of Pinus radiata. The DNA is of sufficiently high quality that it can be amplified by the Polymerase Chain Reaction (PCR). Universal fungal PCR primers were used to amplify fungal material from needles using this method. This was only a small component of the student’s project with the laboratory work scheduled to be completed in September.
The aims of this project are to:
Develop species–specific primers for D. septosporum that can be used to augment traditional diagnostic methods.
Use mating type primers developed by overseas collaborators to distinguish between D. septosporum mating type 2 (already present in New Zealand) and mating type 1 (currently not present in New Zealand).
Commence development of quantitative (real–time) PCR methods to quantify fungal biomass in plant tissue.
Research Provider:
Other
