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On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System
Published on: February 23, 2018
PCR analysis of phytoplasmas based on the secA gene
Matt Dickinson1, Jennifer Hodgetts
1School of Biosciences, University of Nottingham, Sutton Bonington, Nottingham, UK. Matthew.Dickinson@nottingham.ac.uk
Methods in Molecular Biology (Clifton, N.J.)
|September 19, 2012
Summary
Phytoplasma diagnostics and phylogeny can now utilize the secA gene, offering an alternative to the 16S rRNA gene. New universal primers enable secA gene amplification for improved phytoplasma classification.
Area of Science:
- Phytopathology
- Microbiology
- Molecular Biology
Background:
- Phytoplasma diagnostics and phylogenetic analyses traditionally rely on the 16S rRNA gene.
- The development of "universal" primers for alternative genes is crucial for complementary analyses.
Purpose of the Study:
- To detail the use of primers targeting the phytoplasma secA gene.
- To present secA gene-based PCR amplification strategies for phytoplasma identification and phylogeny.
Main Methods:
- Utilizing "universal" primers for the phytoplasma secA gene.
- Employing single or nested PCR approaches for gene amplification.
- Recommending appropriate control measures for PCR assays.
- Leveraging available phytoplasma secA sequences from databases for phylogenetic analysis.
Main Results:
- The secA gene serves as a viable target for phytoplasma diagnostics and phylogenetics.
- Developed primers facilitate amplification of the secA gene in phytoplasmas.
- The methodology supports both single and nested PCR applications.
Conclusions:
- The secA gene offers a valuable complementary tool to the 16S rRNA gene for phytoplasma research.
- This approach enhances the accuracy and scope of phytoplasma classification and identification.
- Standardized protocols using secA gene primers improve molecular diagnostics in phytopathology.
