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Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies
Published on: May 22, 2012
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Direct Polymerase Chain Reaction-Based Detection of Cercospora beticola in Field Soils
R T Lartey1, T C Caesar-TonThat1, A W Lenssen1
1United States Department of Agriculture-Agricultural Research Service (USDA-ARS), Northern Plains Agricultural Laboratory, Sidney, MT 59270.
Plant Disease
|February 13, 2019
Summary
A new polymerase chain reaction (PCR) method rapidly detects Cercospora beticola in soil. This technique aids in managing sugar beet leaf spot by improving disease forecasting and biological control research.
Area of Science:
- Plant pathology
- Molecular biology
- Agricultural science
Background:
- Cercospora beticola causes sugar beet leaf spot, surviving in soil residue.
- Overwintering propagules initiate new infections in sugar beet crops.
Purpose of the Study:
- To develop a rapid polymerase chain reaction (PCR) technique for detecting C. beticola in field soils.
- To enhance research capabilities for sugar beet disease management.
Main Methods:
- DNA extraction from soil using PowerSoil DNA Kit.
- Polymerase chain reaction (PCR) amplification with CBACTIN primers.
- Agarose gel electrophoresis and DNA sequencing for amplicon verification.
Main Results:
- PCR successfully detected C. beticola DNA in amended and field soil samples.
- Amplicon size and sequence alignment confirmed the presence of C. beticola.
- The developed PCR technique showed high specificity and accuracy.
Conclusions:
- The rapid PCR method enables efficient screening of C. beticola in soil.
- This tool will advance research in biological control and disease forecasting.
- Improved management strategies for sugar beet leaf spot are facilitated.
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