Application of fluorescence-based resistance gene analog analysis for genotyping plant genetic resources.
Conventional approaches for detecting disease resistance gene analogs (RGAs) in plants are based on agarose gels or on polyacrylamide gel electrophoresis (PAGE) in combination with silver staining or radioactive labeling. A modified method for RGA analysis has been developed by using fluorescence-labeled primers for PCR amplifications. The amplified fragments are detected by denaturing PAGE using an automated laser fluorescence DNA sequencer and analyzed by fragment analysis software. This technique is not limited to specific plant species and is suitable for high-throughput genotyping plant genetic resources. We demonstrate here the efficiency of this method for comparison of RGA patterns in diverse plant species and for genotyping of natural populations of the wheat progenitor, Triticum dicoccoides.
Cheng
Jianping
J
Institute of Evolution, University of Haifa, Mount Carmel, Haifa, 31905, Israel.
Yan
Jun
J
Krugman
Tamar
T
Fahima
Tzion
T
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005453:Fluorescence; D017343:Genes, Plant; D005819:Genetic Markers; D020022:Genetic Predisposition to Disease; D005828:Genetics, Population; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010802:Phylogeny; D010935:Plant Diseases / Q000235:genetics*; D010944:Plants / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods; D014908:Triticum / Q000235:genetics
D005819:Genetic Markers