Project: 2007-04. Report - DNA-based identification of needle pathogens
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Date: 2007 Author: Dr Rosie Bradshaw and Mr Arne Schwelm Publication: Progress report Project reference: 2007-04 Full report is available from: |
Executive summary:
Commercially available DNA extraction kits are generally as reliable as a “homemade” lab-based method for extracting DNA from needle lesions infected with Dothistroma. However in experiments to determine the minimum number of lesions required for extraction the success was variable with all methods, although sufficient DNA for PCR amplification could be obtained from as few as one or two lesions. A “whole-genome amplification” method is being trialed which may improve success rates with DNA extraction from single lesions. If successful, this method will be especially useful for herbarium samples.PCR primers have been developed that amplify a dothistromin biosynthetic gene. These NZ-developed primers mean we now have three sets of primers which can all be used to identify D. septosporum, but can be used for different purposes and with different levels of resolution (species-specific, mating-type specific, etc.). The dothistromin primers are currently being checked for specificity to ensure they do not detect other common fungi, and real-time PCR experiments are being planned.
Full report is available from:
Rosie Bradshaw
Massey University
Private Bag 11222
Palmerston North
