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Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation

Simple, reliable methods for the identification of alien genetic introgressions are required in plant breeding programmes. The use of genomic dot-blot hybridisation allows the detection of small Hordeum chilense genomic introgressions in the descendants of genetic crosses between wheat and H. chilen...

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Autores principales: Rey, María -Dolores, Prieto, Pilar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5344947/
https://www.ncbi.nlm.nih.gov/pubmed/28337069
http://dx.doi.org/10.1007/s11032-017-0629-5
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author Rey, María -Dolores
Prieto, Pilar
author_facet Rey, María -Dolores
Prieto, Pilar
author_sort Rey, María -Dolores
collection PubMed
description Simple, reliable methods for the identification of alien genetic introgressions are required in plant breeding programmes. The use of genomic dot-blot hybridisation allows the detection of small Hordeum chilense genomic introgressions in the descendants of genetic crosses between wheat and H. chilense addition or substitution lines in wheat when molecular markers are difficult to use. Based on genomic in situ hybridisation, DNA samples from wheat lines carrying putatively H. chilense introgressions were immobilised on a membrane, blocked with wheat genomic DNA and hybridised with biotin-labelled H. chilense genomic DNA as a probe. This dot-blot screening reduced the number of plants necessary to be analysed by molecular markers or in situ hybridisation, saving time and money. The technique was sensitive enough to detect a minimum of 5 ng of total genomic DNA immobilised on the membrane or about 1/420 dilution of H. chilense genomic DNA in the wheat background. The robustness of the technique was verified by in situ hybridisation. In addition, the detection of other wheat relative species such as Hordeum vulgare, Secale cereale and Agropyron cristatum in the wheat background was also reported.
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spelling pubmed-53449472017-03-21 Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation Rey, María -Dolores Prieto, Pilar Mol Breed Article Simple, reliable methods for the identification of alien genetic introgressions are required in plant breeding programmes. The use of genomic dot-blot hybridisation allows the detection of small Hordeum chilense genomic introgressions in the descendants of genetic crosses between wheat and H. chilense addition or substitution lines in wheat when molecular markers are difficult to use. Based on genomic in situ hybridisation, DNA samples from wheat lines carrying putatively H. chilense introgressions were immobilised on a membrane, blocked with wheat genomic DNA and hybridised with biotin-labelled H. chilense genomic DNA as a probe. This dot-blot screening reduced the number of plants necessary to be analysed by molecular markers or in situ hybridisation, saving time and money. The technique was sensitive enough to detect a minimum of 5 ng of total genomic DNA immobilised on the membrane or about 1/420 dilution of H. chilense genomic DNA in the wheat background. The robustness of the technique was verified by in situ hybridisation. In addition, the detection of other wheat relative species such as Hordeum vulgare, Secale cereale and Agropyron cristatum in the wheat background was also reported. Springer Netherlands 2017-03-09 2017 /pmc/articles/PMC5344947/ /pubmed/28337069 http://dx.doi.org/10.1007/s11032-017-0629-5 Text en © The Author(s) 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Article
Rey, María -Dolores
Prieto, Pilar
Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation
title Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation
title_full Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation
title_fullStr Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation
title_full_unstemmed Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation
title_short Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation
title_sort detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5344947/
https://www.ncbi.nlm.nih.gov/pubmed/28337069
http://dx.doi.org/10.1007/s11032-017-0629-5
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