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Rapid gene isolation in barley and wheat by mutant chromosome sequencing

Identification of causal mutations in barley and wheat is hampered by their large genomes and suppressed recombination. To overcome these obstacles, we have developed MutChromSeq, a complexity reduction approach based on flow sorting and sequencing of mutant chromosomes, to identify induced mutation...

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Detalles Bibliográficos
Autores principales: Sánchez-Martín, Javier, Steuernagel, Burkhard, Ghosh, Sreya, Herren, Gerhard, Hurni, Severine, Adamski, Nikolai, Vrána, Jan, Kubaláková, Marie, Krattinger, Simon G., Wicker, Thomas, Doležel, Jaroslav, Keller, Beat, Wulff, Brande B. H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5087116/
https://www.ncbi.nlm.nih.gov/pubmed/27795210
http://dx.doi.org/10.1186/s13059-016-1082-1
Descripción
Sumario:Identification of causal mutations in barley and wheat is hampered by their large genomes and suppressed recombination. To overcome these obstacles, we have developed MutChromSeq, a complexity reduction approach based on flow sorting and sequencing of mutant chromosomes, to identify induced mutations by comparison to parental chromosomes. We apply MutChromSeq to six mutants each of the barley Eceriferum-q gene and the wheat Pm2 genes. This approach unambiguously identified single candidate genes that were verified by Sanger sequencing of additional mutants. MutChromSeq enables reference-free forward genetics in barley and wheat, thus opening up their pan-genomes to functional genomics. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13059-016-1082-1) contains supplementary material, which is available to authorized users.