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 smallHordeum chilensegenomic introgressions in the descendants of genetic crosses between wheat andH. chilenseaddition or substitution lines in wheat when molecular markers are difficult to use. Based on genomic in situ hybridisation, DNA samples from wheat lines carrying putativelyH. chilenseintrogressions were immobilised on a membrane, blocked with wheat genomic DNA and hybridised with biotin-labelledH. chilensegenomic 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 ofH. chilensegenomic 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 asHordeum vulgare,Secale cerealeandAgropyron cristatumin the wheat background was also reported.
Plant Breeding Department, Institute for Sustainable Agriculture, Agencia Estatal Consejo Superior de Investigaciones Científicas (CSIC), Alameda del Obispo s/n, 14080 Córdoba, Spain.
María-Dolores
MD
Rey
Plant Breeding Department, Institute for Sustainable Agriculture, Agencia Estatal Consejo Superior de Investigaciones Científicas (CSIC), Alameda del Obispo s/n, 14080 Córdoba, Spain.
Pilar
0000-0002-8160-808X
P
Prieto
Bread wheat; Dot-blot hybridisation; Genomic introgressions; H. chilense