Home › Botany › On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System
Botany JoVE (Open Access) Citable · DOI

On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System

DOI: 10.3791/56891-v
What you'll learn
  • ✓ Extract DNA from soil samples using magnetic bead-based purification
  • ✓ Perform soil lysis for phytopathogen detection
  • ✓ Operate portable real-time PCR systems in field conditions
  • ✓ Diagnose soil-borne plant pathogens on-site
Protocol

Rapid and accurate detection of plant pathogens on-site, especially soil-borne pathogens, is crucial to prevent further inoculum production and proliferation of plant diseases in the field. The method developed here using a portable real-time PCR detection system enables on-site diagnosis under field conditions.

Difficulty
intermediate
Total time
~1–2 hours per soil sample
Biosafety
BSL-1

Steps

1
Extract DNA using magnetic bead-based method

Isolate pathogenic DNA from soil samples using magnetic bead purification. This approach enables rapid nucleic acid extraction suitable for field deployment.

▶ 01:06
2
Lyse soil samples for pathogen release

Prepare soil lysate through lysis steps to release phytopathogen cells and DNA from soil matrix. This step is critical for enabling downstream PCR detection.

▶ 02:06
3
Analyze results and interpret pathogen detection

Review representative real-time PCR amplification curves and quantification data to confirm presence or absence of soil-borne pathogens in field samples.

▶ 11:44
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