Rapid verification of wheat-Hordeum introgressions by direct staining of SCAR, STS, and SSR amplicons.

A range of single tagged site (STS), simple sequence repeat (SSR), and sequence-characterized amplified region (SCAR) markers were screened for their utility in detecting Hordeum vulgare and H. chilense chromosomes in a wheat background. PCR conditions were optimized for specific amplification of the targeted sequences and to avoid cross-species amplification. Two H. vulgare derived STSs, six H. vulgare derived SSRs, and nine H. chilense derived SCARs were usable for the detection of five H. vulgare and three H. chilense chromosomes by direct ethidium bromide staining of the PCR products in test tubes, avoiding the more costly and time-consuming DNA electrophoresis step. The practical application of the method is illustrated by the identification of a monotelosomic substitution of H. vulgare chromosome 6HS in tritordeum and a monosomic addition of H. chilense chromosome 6Hch in durum wheat.

[1]  P. Hernández,et al.  Development of SCARs by direct sequencing of RAPD products: a practical tool for the introgression and marker-assisted selection of wheat , 1999, Molecular Breeding.

[2]  A. Cabrera,et al.  Application of C-banding and fluorescence in situ hybridization for the identification of the trisomics of Hordeum chilense , 1999, Euphytica.

[3]  M. Maroof,et al.  Development of simple sequence repeat DNA markers and their integration into a barley linkage map , 1996, Theoretical and Applied Genetics.

[4]  P. Langridge,et al.  STS-PCR markers appropriate for wheat-barley introgression , 1996, Theoretical and Applied Genetics.

[5]  N. Weeden,et al.  Large-scale, cost-effective screening of PCR products in marker-assisted selection applications , 1995, Theoretical and Applied Genetics.

[6]  K. Shepherd,et al.  Production of wheat-barley recombinant chromosomes through induced homoeologous pairing , 1992, Theoretical and Applied Genetics.

[7]  D. Laurie,et al.  Utility of barley and wheat simple sequence repeat (SSR) markers for genetic analysis of Hordeum chilense and tritordeum , 2002, Theoretical and Applied Genetics.

[8]  D. Laurie,et al.  A core genetic map of Hordeum chilense and comparisons with maps of barley (Hordeum vulgare) and wheat (Triticum aestivum) , 2001, Theoretical and Applied Genetics.

[9]  A. Martín,et al.  Development and characterization of Hordeum chilense chromosome-specific STS markers suitable for wheat introgression and marker-assisted selection , 1999, Theoretical and Applied Genetics.

[10]  N. Blake,et al.  Transfer of sequence tagged site PCR markers between wheat and barley. , 1996, Genome.

[11]  P. Hernández,et al.  Development of RAPD markers in tritordeum and addition lines of Hordeum chilense in Triticum aestivum , 1996 .

[12]  M. Heun,et al.  Mapping of digested and undigested random amplified microsatellite polymorphisms in barley. , 1995, Genome.

[13]  P. Walsh,et al.  Simultaneous Amplification and Detection of Specific DNA Sequences , 1992, Bio/Technology.

[14]  W. F. Thompson,et al.  Rapid isolation of high molecular weight plant DNA. , 1980, Nucleic acids research.

[15]  W. Gerlach,et al.  Cloning and characterization of ribosomal RNA genes from wheat and barley. , 1979, Nucleic acids research.