IMPROVEMENT AND DEPLOYMENT OF RAPID STANDARDIZED PCR DIAGNOSTIC TOOLS TO INCREASE DETECTION CAPACITY FOR HIGH-IMPACT PLANT PATHOGEMS
Cereal Disease Laboratory
2012 Annual Report
1a.Objectives (from AD-416):
The long-term goal of this integrated project is to increase early and accurate detection of high-impact plant pathogens. The specific objectives are:.
1)Research – Adaptation of existing plant pathogen protocols for use in diagnostic laboratories and the production of high-quality, standardized controls;.
2)Extension – Develop and disseminate disease-specific sample submission information to increase the quality and quantity of important plant disease samples;.
3)Evaluation – Determine the success and impact of the research and extension portions by using surveys of end-users and assessment of data generated by research and extension projects.
1b.Approach (from AD-416):
This project will focus on developing or adapting diagnostic protocols for the following high-impact plant pathogens:.
2)Puccinia graminis f.sp. tritici;.
4)Candidatus Liberibacter asiaticus;.
5)three major virus genera (Crinivirus, Begomovirus and Potyvirus). Available diagnostic protocols based on PCR will be tested and modified where necessary in order to adapt to a plant diagnostic clinic setting. Positive controls will be developed for each of the assays. DNA extractions methods will be improved and modified for efficient and high-throughput laboratories. Internal controls will be developed to verify assay conditions. Bead-based PCR reagents will be developed. Assays will be verified in test laboratories. Training activities (workshops, development of online materials and printed materials) will be developed and deployed for diagnosticians.
This project is part of a larger program to develop and deploy a rapid molecular diagnostic assay for the Ug99 race group of the wheat stem rust pathogen (Puccinia graminis f. sp. tritici). A DNA based assay was developed based on four independent target regions of the genome. Specificity and performance of the assay was tested on global set of samples. In addition, testing in three independent laboratories validated the performance of the assay.