Project Details:

Quantification of disease severity of charcoal rot in plant hosts by qPCR

Parent Project: This is the first year of this project.
Checkoff Organization:Kansas Soybean Commission
Categories:Soybean diseases, Analytical standards & measurements
Organization Project Code:
Project Year:2015
Lead Principal Investigator:Bin Shuai (Wichita State University)
Co-Principal Investigators:
Keywords: Charcoal Rot, Disease Diagnosis, Research Methodology

Contributing Organizations

Funding Institutions

Information and Results

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Project Summary

Charcoal rot is a disease caused by the soil-borne necrotrophic fungus Macrophomina phaseolina (M. phaseolina). The pathogen has a very wide host range and affects plant growth, yield and seed quality. Currently, there is no effective management approach to control the disease and disease resistance cultivars are not available for most crop species. To identify potential host genes for engineering resistant cultivars, our laboratory has established two pathosystems using model plants Medicago truncatula and Arabidopsis thaliana to study the molecular interactions between the pathogen and its plant hosts. To characterize the disease symptoms and progression, we established a scoring system based on percentage of necrosis and chlorosis that appeared on the aerial part of the plants. This system provides a quick and easy way to assess the disease progression, and it is commonly applied in studying many types of plant diseases.

Project Objectives

1. Establish a qPCR assay that can accurately quantify the amount of M. phaseolina in different types of samples and is suitable for disease progression analysis in host plants.
2. Test and optimize qPCR conditions to achieve accurate quantification of M. phaseolina DNA.
3. Apply the assay to evaluate disease progression in two pathosystems, Medicago truncatula and Arabidopsis thaliana, as models to study charcoal rot disease.

Project Deliverables

Progress of Work

Final Project Results


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Using a series dilutions of M. phaseolina genomic DNA in a range of 5 log difference in concentrations, we determined that primers for A1 and A2 amplicons are suitable for further study. However, primers for A2 amplicon were more sensitive and were able to amplify small amount of template. A protocol has been established to successfully isolate high quality genomic DNA from M. phaseolina, however, the protocol needs to be fine-tuned in order for it to work for infected plant tissues. My graduate student, Prajwal Bhandari, is working on optimizing the DNA extraction protocol to achieve consistent results. Currently, a graduate student is working on optimizing the protocol for genomic DNA isolation from infected plant samples and the real-time PCR assay. The assay will be applied to our working pathosystems in short term. During the grant period, two graduate students were supported as GRAs. One of them graduated in Summer and is admitted to a Ph.D program at University of Iowa. The other graduate student continues to work on the project and also develops new project based on current findings.

Benefit to Soybean Farmers

Performance Metrics

Project Years