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Theoretical and Applied Genetics

, Volume 96, Issue 3–4, pp 371–375 | Cite as

A simple PCR-based method for scoring the ph1b deletion in wheat

  • L.-J. Qu
  • T. N. Foote
  • M. A. Roberts
  • T. A. Money
  • L. Aragón-Alcaide
  • J. W. Snape
  • G. Moore
Article

Abstract

The amplified fragment length polymorphism (AFLP) technique was used to isolate DNA sequences present in the euploid wheat Chinese Spring but not in the Chinese Spring ph1b mutant (which has a deletion of the Ph1 gene, a suppressor of homoeologous chromosome pairing). The polymorphic DNA fragments identified by AFLP were then cloned, sequenced, and used to design two primer pairs. These primers were used in a PCR-based assay to specifically amplify products from the Chinese Spring euploid but not from the ph1b mutant. This PCR assay can be carried out from extracted genomic DNA or directly from alkaline-treated wheat leaves, and the reaction products can be scored on a plus-minus basis, making the screening amenable to automation. The reliability of the assay was tested using a F1-derived doubled-haploid population of 55 lines which segregate for the ph1b deletion. This PCR-screening technique is less time and labour consuming, and more accurate and reliable, than cytologically based conventional methods.

Key words AFLP Chinese Spring Chromosome pairing PCR assay ph1b deletion 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • L.-J. Qu
    • 1
  • T. N. Foote
    • 1
  • M. A. Roberts
    • 1
  • T. A. Money
    • 1
  • L. Aragón-Alcaide
    • 1
  • J. W. Snape
    • 1
  • G. Moore
    • 1
  1. 1.Cereals Research Department, John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, UKGB

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