Development of rapid and sensitive molecular diagnostic tools to detect trunk disease pathogens from environmental and plant materials

Abstract

Dr. Regina Billones-Baaijens travelled to Canada in March – June, 2016 to undergo training on new molecular techniques and conduct collaborative research with Dr. Jose R. Úrbez Torres at the Summerland Research and Development Centre (SuRDC), Agriculture and Agri-Food Canada, Summerland, British Columbia. This travel was financially supported by a National Wine and Grape Industry Centre (NWGIC) Research Fellowship Grant; a Wine Australia Travel Bursary with in-kind contribution from SuRDC. The output of this travel will contribute to one of the objectives of a current Wine Australia-funded project, SAR1205 ‘Practical management of grapevine trunk diseases (GTD)' and the newly approved project, SAR1601 'Grapevine trunk disease management for vineyard longevity in diverse climates of Australia'. These projects aim to ascertain spore dispersal of GTD pathogens and to determine the climatic conditions that favour the release of these pathogens. The new project also aims to quantify levels of GTD pathogens in nursery propagation material and determine different stress conditions that trigger disease expression in vineyards. The three month collaborative research period at SuRDC included hands-on training on (a) DNA macroarray; (b) Droplet Digital PCR (ddPCR); and (c) specificity test and qPCR optimisation of Eutypa dieback (ED) and Botryosphaeria dieback (BD) multi-species primers developed for the GTD project (SAR1205). The DNA macroarray and ddPCR are rapid and specific tools for the identification, detection and quantification of plant pathogens. The training at SuRDC has resulted in enhanced skills in the development of rapid and accurate molecular tools for the detection and quantification of GTD pathogens in the environment. In addition, experience was gained to optimise and troubleshoot molecular assays that will fast track the analyses of spore trap samples collected for the spore trapping studies in Australia. The skills gained in Canada will allow researchers at the NWGIC to adopt these DNA-based diagnostic tools for future GTD research subject to the availability of equipment. The transfer of technologies between Canadian and Australian researchers builds on collaboration established through previous exchanges between the countries by Drs Urbez Torres and Mark Sosnowski (SARDI) and will contribute to the development of control strategies for grapevine trunk diseases that is considered a serious threat to the sustainability of both wine industries.

Summary

The travel to Canada has allowed Dr Billones-Baaijens to enhance her skills in the development of rapid and accurate molecular tools for the detection of GTD pathogens in the environment. It has further enhanced her skills in optimisation and troubleshooting of molecular assays that will fast track the analyses of spore trap samples collected from different wine growing regions enabling the identification of critical periods for spore release. This will allow the development of industry recommendations particularly in pruning practices and fungicide applications to prevent infections by these pathogens. This travel also contributed in the transfer of technologies and exchange of GTD isolates between Canada and Australia and to the development of control strategies for grapevine trunk diseases that threaten the sustainability of the wine industry for both countries.

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