Identification of barley mutants in the cultivar ‘Lux’ at the Dhn loci through TILLING

Authors

  • S. Lababidi,

    1. Department of Agriculture and Ecology, Plant and Soil Science Laboratory, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark
    2. Biotechnology Laboratory, International Center for Agricultural Research in the Dry Areas, Aleppo, Syria
    3. Department of Plant Science, Life Science Faculty, Aleppo University, Aleppo, Syria
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  • N. Mejlhede,

    1. Department of Agriculture and Ecology, Plant and Soil Science Laboratory, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark
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  • S. K. Rasmussen,

    1. Department of Agriculture and Ecology, Plant and Soil Science Laboratory, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark
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  • G. Backes,

    1. Department of Agriculture and Ecology, Plant and Soil Science Laboratory, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark
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  • W. Al-Said,

    1. Department of Plant Science, Life Science Faculty, Aleppo University, Aleppo, Syria
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  • M. Baum,

    1. Biotechnology Laboratory, International Center for Agricultural Research in the Dry Areas, Aleppo, Syria
    2. Corresponding author, E-mail: m.baum@cgiar.org
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  • A. Jahoor

    1. Department of Agriculture and Ecology, Plant and Soil Science Laboratory, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark
    2. Nordic Seed, Kløngevej 6, DK-4983 Dannemare, Denmark
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Abstract

TILLING is a reverse genetic strategy that allows screening for mutations in genes with known sequences in a plant mutant population. A TILLING population has been developed for the Danish barley variety ‘Lux’ (Hordeum vulgare L.), by using sodium azide to induce mutations. Scoring of four visible phenotypic characters of barley seedling in reference to the parental cultivar ‘Lux’ in the M3 plants showed over 3.5% lethality. A series of pool ratios of mixed DNA from mutant lines were tested and 10-fold pools appeared to be the practical mixing ratio for the detection of fragments in the 500–700 bp range. Two of the 13 known dehydrin genes, Dhn12 and Dhn13, respectively, were examined and five independent missense mutations were obtained from a population of 9575 barley mutant plants. This corresponds to a mutation density of approximately one mutation every two and half million base pairs for these two genes. The mutant population of approximately 10 000 lines was screened for mutations in two genes in a short time due to high pooling ratio.

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