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Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions
Published on: September 30, 2016
Random amplified polymorphic DNA analysis of Ustilago violacea
C E Henry1, P Okoro, E Steward-Clark
1Chicago State University, Department of Biological Sciences, Illinois 60628, USA. C-Henry@csu.edu
Annals of the New York Academy of Sciences
|July 23, 1999
Summary
Arbitrarily primed polymerase chain reaction (PCR) effectively identified genetic differences between Ustilago violacea strains. This molecular tool aids in detecting transposable elements and understanding fungal genetics.
Area of Science:
- Mycology
- Plant Pathology
- Molecular Biology
Background:
- Ustilago violacea, a basidiomycete phytopathogen, exhibits a narrow host range within the Caryophyllaceae family.
- Traditional genetic mapping methods include RFLPs, electrophoretic karyotypes, and CHEF gel analysis.
- Arbitrarily primed PCR offers a powerful method for genomic fingerprinting and strain differentiation.
Purpose of the Study:
- To evaluate the efficiency of PCR with specific primers in detecting intraspecific genetic variation in Ustilago violacea.
- To differentiate between the two mating type strains (a1 and a2) of U. violacea using PCR-based genomic fingerprinting.
Main Methods:
- Utilized PCR technology with 40 Operon 10-mer primers and 5 simple sequence repeat (SSR) primers.
- Analyzed DNA samples from U. violacea mating type strains.
- Detected DNA polymorphisms via banding patterns after PCR and 1.4% agarose gel electrophoresis.
Main Results:
- PCR successfully generated polymorphic banding patterns, indicating intraspecific genetic variation between U. violacea strains.
- The chosen primers effectively distinguished between the genomic DNA of the a1 and a2 mating types.
Conclusions:
- PCR-based genomic fingerprinting is an efficient method for detecting intraspecific variation in Ustilago violacea.
- This identified polymorphism will be instrumental in detecting cryptic and trans-active transposable elements within the species.
