2015
Quantification of disease severity of charcoal rot in plant hosts by qPCR
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Contributor/Checkoff:
Kansas Soybean Commission
Category:
Sustainable Production
Keywords:
Crop protectionDiseaseField management
Parent Project:
This is the first year of this project.
Lead Principal Investigator:
Bin Shuai, Wichita State University
Co-Principal Investigators:
Project Code:
Contributing Organization (Checkoff):
Kansas Soybean Commission
(n/a)
Institution Funded:
Kansas State University
(n/a)
Brief Project Summary:

Charcoal rot is a disease caused by the soil-borne necrotrophic fungus Macrophomina phaseolina. To identify potential host genes for engineering resistant cultivars, our laboratory has established two patho-systems using model plants to study the molecular interactions between the pathogen and its plant hosts. Particular objectives include: establishing 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; testing and optimizing qPCR conditions to achieve accurate quantification of M. phaseolina DNA; and applying the assay to evaluate disease progression in two pathosystems to study charcoal rot.

Key Benefactors:
farmers, plant pathologists, agronomists, breeders

Information And Results
Project Deliverables

Final Project Results

Update:

View uploaded report Word file

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.

The United Soybean Research Retention policy will display final reports with the project once completed but working files will be purged after three years. And financial information after seven years. All pertinent information is in the final report or if you want more information, please contact the project lead at your state soybean organization or principal investigator listed on the project.