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Barley necrotic locus nec1 encodes the cyclic nucleotide-gated ion channel 4 homologous to the Arabidopsis HLM1

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Abstract

Barley homolog of the Arabidopsis necrotic (disease lesion mimic) mutant HLM1 that encodes the cyclic nucleotide-gated ion channel 4 was cloned. Barley gene was mapped genetically to the known necrotic locus nec1 and subsequent sequence analysis identified mutations in five available nec1 alleles confirming barley homolog of Arabidopsis HLM1 as the NEC1 gene. Two fast neutron (FN) induced mutants had extensive deletions in the gene, while two previously described nec1 alleles had either a STOP codon in exon 1 or a MITE insertion in intron 2 which caused alternative splicing, frame shift and production of a predicted non-functional protein. The MITE insertion was consistent with the reported spontaneous origin of the nec1 Parkland allele. The third FN mutant had a point mutation in the coding sequence which resulted in an amino acid change in the conserved predicted cyclic nucleotide-gated ion channel pore region. The expression of two pathogenesis-related genes, HvPR-1a and β-1,3-glucanase, was elevated in two FN necrotic lines. Ten other members of the barley cyclic nucleotide-gated ion channel gene family were identified and their position on barley linkage map is reported.

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Acknowledgements

Research was supported by USDA/NRI grant No. 9901325 to AK and BBSRC/SEERAD grant SCR/903/02 to RW. We thank Dr. An Hang for the seed of nec1 alleles in Carlsberg II and Parkland, as well as the nec1 backcross to Bowman, Dr. Harold Bockelman for the seed of respective parents, Maura Lyons for help with identification of the MITE insertion, Dr. Alison Roberts for help with fluorescence microscopy and Drs. Ingo Hein and Jerome Franckowiak for helpful discussions. Technical assistance from Niki McCallum in screening the Optic mutant population is gratefully acknowledged.

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Authors and Affiliations

  1. Scottish Crop Research Institute, Genome Dynamics, Invergowrie, DD2 5DA, Dundee, Scotland, UK

    Nils Rostoks, Sharon Mudie, David G. Caldwell & Robbie Waugh

  2. Department of Crop and Soil Sciences, Washington State University, Pullman, WA, 99164, USA

    Deric Schmierer, Thomas Drader, Robert Brueggeman & Andris Kleinhofs

  3. School of Molecular Biosciences, Washington State University, Pullman, WA, 99164, USA

    Andris Kleinhofs

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  1. Nils Rostoks
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  2. Deric Schmierer
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  3. Sharon Mudie
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  5. Robert Brueggeman
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  6. David G. Caldwell
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  7. Robbie Waugh
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  8. Andris Kleinhofs
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Correspondence to Nils Rostoks.

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Communicated by R. Hagemann

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Rostoks, N., Schmierer, D., Mudie, S. et al. Barley necrotic locus nec1 encodes the cyclic nucleotide-gated ion channel 4 homologous to the Arabidopsis HLM1. Mol Genet Genomics 275, 159–168 (2006). https://doi.org/10.1007/s00438-005-0073-9

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  • DOI: https://doi.org/10.1007/s00438-005-0073-9

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