Rapid gene isolation in barley and wheat by mutant chromosome sequencing

Javier Sánchez-Martín; Burkhard Steuernagel; Sreya Ghosh; Gerhard Herren; Severine Hurni; Nikolai Adamski; Jan Vrána; Marie Kubaláková; Simon Krattinger; Thomas Wicker; Jaroslav Doležel; Beat Keller; Brande B.H. Wulff

doi:10.1186/s13059-016-1082-1

Rapid gene isolation in barley and wheat by mutant chromosome sequencing

Javier Sánchez-Martín, Burkhard Steuernagel, Sreya Ghosh, Gerhard Herren, Severine Hurni, Nikolai Adamski, Jan Vrána, Marie Kubaláková, Simon Krattinger, Thomas Wicker, Jaroslav Doležel, Beat Keller^*, Brande B.H. Wulff

^*Corresponding author for this work

Biological, Environmental Science and Engineering

Research output: Contribution to journal › Article › peer-review

97 Scopus citations

Abstract

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.

Original language	English (US)
Article number	221
Journal	Genome biology
Volume	17
Issue number	1
DOIs	https://doi.org/10.1186/s13059-016-1082-1
State	Published - Oct 31 2016

Keywords

Barley
Chromosome flow sorting
Gene cloning
MutChromSeq
Mutational genomics
Triticeae
Wheat

ASJC Scopus subject areas

Ecology, Evolution, Behavior and Systematics
Genetics
Cell Biology

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abstract = "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.",

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doi = "10.1186/s13059-016-1082-1",

language = "English (US)",

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Rapid gene isolation in barley and wheat by mutant chromosome sequencing. / Sánchez-Martín, Javier; Steuernagel, Burkhard; Ghosh, Sreya; Herren, Gerhard; Hurni, Severine; Adamski, Nikolai; Vrána, Jan; Kubaláková, Marie; Krattinger, Simon; Wicker, Thomas; Doležel, Jaroslav; Keller, Beat; Wulff, Brande B.H.

In: Genome biology, Vol. 17, No. 1, 221, 31.10.2016.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Rapid gene isolation in barley and wheat by mutant chromosome sequencing

AU - Sánchez-Martín, Javier

AU - Steuernagel, Burkhard

AU - Ghosh, Sreya

AU - Herren, Gerhard

AU - Hurni, Severine

AU - Adamski, Nikolai

AU - Vrána, Jan

AU - Kubaláková, Marie

AU - Krattinger, Simon

AU - Wicker, Thomas

AU - Doležel, Jaroslav

AU - Keller, Beat

AU - Wulff, Brande B.H.

PY - 2016/10/31

Y1 - 2016/10/31

N2 - 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.

AB - 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.

KW - Barley

KW - Chromosome flow sorting

KW - Gene cloning

KW - MutChromSeq

KW - Mutational genomics

KW - Triticeae

KW - Wheat

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Rapid gene isolation in barley and wheat by mutant chromosome sequencing

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Keywords

ASJC Scopus subject areas

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