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Rapid gene isolation in barley and wheat by mutant chromosome sequencing


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.

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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27 downloads since deposited on 12 Jan 2017
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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Plant and Microbial Biology
Dewey Decimal Classification:580 Plants (Botany)
Language:English
Date:31 October 2016
Deposited On:12 Jan 2017 09:23
Last Modified:23 Sep 2018 06:05
Publisher:BioMed Central
ISSN:1474-7596
OA Status:Gold
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1186/s13059-016-1082-1
PubMed ID:27795210
Project Information:
  • : FunderSNSF
  • : Grant ID310030_163260
  • : Project TitleMolecular analysis of three different types of fungal disease resistance mechanisms in cereals
  • : FunderSNSF
  • : Grant IDPZ00P3_154694
  • : Project TitleFunctional characterization of broad-spectrum disease resistance in cereals
  • : FunderSNSF
  • : Grant ID310030B_144081
  • : Project TitleMolecular interactions of wheat and fungal pathogens: Lr34-based, durable resistance and the wheat-powdery mildew pathosystem
  • : FunderSNSF
  • : Grant ID310030_163260
  • : Project TitleMolecular analysis of three different types of fungal disease resistance mechanisms in cereals
  • : FunderSNSF
  • : Grant IDPZ00P3_154694
  • : Project TitleFunctional characterization of broad-spectrum disease resistance in cereals
  • : FunderSNSF
  • : Grant ID310030B_144081
  • : Project TitleMolecular interactions of wheat and fungal pathogens: Lr34-based, durable resistance and the wheat-powdery mildew pathosystem
  • : FunderSNSF
  • : Grant ID310030_163260
  • : Project TitleMolecular analysis of three different types of fungal disease resistance mechanisms in cereals
  • : FunderSNSF
  • : Grant IDPZ00P3_154694
  • : Project TitleFunctional characterization of broad-spectrum disease resistance in cereals
  • : FunderSNSF
  • : Grant ID310030B_144081
  • : Project TitleMolecular interactions of wheat and fungal pathogens: Lr34-based, durable resistance and the wheat-powdery mildew pathosystem

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