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

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Abstract:

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.

Un ensemble de marquers de type STS (« single-tagged-site »), microsatellite (SSR) et SCAR (« sequence-characterized amplified region ») ont été examinés quant à leur utilité potentielle pour la détection de chromosomes du Hordeum vulgare et du H. chilense dans un fond génétique du blé. Les conditions PCR ont été optimisées pour l'amplification spécifique des séquences ciblées et pour éviter l'amplification croisée entre espèces. Deux marqueurs STS dérivés du H. vulgare, six microsatellites dérivés du H. vulgare et neuf SCAR dérivés du H. chilense permettaient de détecter cinq chromosomes du H. vulgare et trois chromosomes du H. chilense par simple détection des amplicons au bromure d'éthidium dans des tubes réactionnels, évitant ainsi l'étape plus longue et coûteuse de l'électrophorèse. L'intérêt pratique de cette méthode est illustré via l'identification d'une substitution monotélosomique du chromosome 6HS du H. vulgare chez une lignée issue d'un croisement blé-orge (tritordeum) ainsi que par l'identification d'une addition monosomique du chromosome 6Hch du H. chilense chez le blé durum.Mots clés : GISH, Hordeum chilense, Hordeum vulgare, sélection assistée, microsatellite, Tritordeum[Traduit par la Rédaction]

Keywords: GISH; Hordeum chilense; Hordeum vulgare; SSR; Tritordeum; marker-assisted selection; microsatellite; sélection assistée

Document Type: Research Article

Publication date: February 1, 2002

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  • From its inception in 1957, this international cytogenetics journal has catered to the research areas of the members of the Genetics Society of Canada; traditionally, these have included agriculture, entomology, genetics/cytogenetics, and evolutionary mechanisms. The contents of the journal have evolved as contributors developed new technologies and interests. A 20-member Editorial Board is composed of scientists from around the world. Reviews and commentary from respected experts are often featured.
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nrc/gen/2002/00000045/00000001/art00025
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