Application of fluorescence-based resistance gene analog analysis for genotyping plant genetic resources

Jianping Cheng, Jun Yan, Tamar Krugman, Tzion Fahima

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)83-89
Number of pages7
JournalBiotechnology Letters
Volume27
Issue number2
DOIs
StatePublished - Jan 2005

Bibliographical note

Funding Information:
This study was supported by the German–Israeli Cooperation Project (DIP project No. DIP-B 4.3), funded by the BMBF and supported by BMBF’s International Bureau at the DLR, and the ISRAEL SCIENCE FOUNDATION (No. 9030/96 and 9048/99). The authors wish to thank Dr Marion Röder, at the Institute of Plant Genetics and Crop Plant Research (IPK), Germany, for her kind help and critical reading of the manuscript.

Keywords

  • DNA fingerprinting
  • Fluorescence-based fragment analysis
  • Resistance gene analog

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Bioengineering
  • Biotechnology

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