Rapid verification of wheat Hordeum introgressions by direct staining of SCAR, STS, and SSR amplicons
[EN] 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.Key words: GISH, Hordeum chilense, Hordeum vulgare, marker-assisted selection, SSR, Tritordeum.
| Main Authors: | , , , , , |
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| Other Authors: | |
| Format: | artículo biblioteca |
| Language: | English |
| Published: |
National Research Council Canada
2002
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| Subjects: | GISH, Hordeum chilense, Hordeum vulgare, Marker-assisted selection, SSR, Tritordeum, Sélection assistée, Microsatellites, |
| Online Access: | http://hdl.handle.net/10261/184200 http://dx.doi.org/10.13039/501100006280 http://dx.doi.org/10.13039/501100007273 |
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| Summary: | [EN] 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.Key words: GISH, Hordeum chilense, Hordeum vulgare, marker-assisted selection, SSR, Tritordeum. |
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