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Identification of candidate CBF genes for the frost tolerance locus Fr-A m 2 in Triticum monococcum

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Abstract

A cluster of eleven CBF genes was recently mapped to the Frost resistance-2 (Fr-A m 2) locus on chromosome 5 of diploid wheat (Triticum monococcum) using a cross between frost tolerant accession G3116 and frost sensitive DV92. The Fr-A m 2 locus was mapped at the peak of two overlapping quantitative trait loci (QTL), one for frost survival and the other for differential expression of the cold regulated gene COR14b. Seven lines with recombination events within the CBF cluster were used to identify CBF candidate genes for these QTL. The lines carrying the critical recombination events were tested for whole plant frost survival and for differential transcript levels of cold induced COR14b and DHN5 genes. The strongest effect for these traits was associated to the linked TmCBF12, TmCBF14 and TmCBF15 genes, with the G3116 allele conferring improved frost tolerance and higher levels of COR14b and DHN5 transcript at mild cold temperatures (12–15°C) than the DV92 allele. Comparison of CBF protein sequences revealed that the DV92 TmCBF12 protein contains a deletion of five amino acids in the AP2 DNA binding domain. Electrophoretic Mobility Shift Assays (EMSA) confirmed that the protein encoded by this allele cannot bind to the CRT/DRE (C-repeat/dehydration-responsive element) motif present in the promoters of several cold induced genes. A smaller effect on frost tolerance was mapped to the distal group of CBF genes including TmCBF16. Transcript levels of TmCBF16, as well as those of TmCBF12 and TmCBF15 were up-regulated at mild cold temperatures in G3116 but not in DV92. Higher threshold induction temperatures can result in earlier initiation of the cold acclimation process and better resistance to subsequent freezing temperatures. The non-functional TmCBF12 allele in DV92 can also contribute to its lower frost tolerance.

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Abbreviations

bp:

Base pair

CBF :

C-Repeat binding factor

COR :

Cold regulated

CRT/DRE:

C-Repeat/Dehydration response element

DHN :

Dehydrin

Fr :

Frost resistance

QTL:

Quantitative trait locus

SEM:

Standard errors of the means

SNP:

Single nucleotide polymorphism

SOM:

Supplementary online material

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Acknowledgements

This project was supported by the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number 2004-01783, the Hungarian Wheat Spike Consortia (NKFP 406404) and the Hungarian National Research Fund No. T046573. Work in the Stockinger lab is supported in part by grants from the NSF Plant Genome Program (DBI 0110124) and the Ohio Plant Biotechnology Consortium (Proposal Number 2007-020). The Hungarian authors thank Alexandra Soltész for her technical assistance.

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Authors and Affiliations

  1. Department of Horticulture and Crop Science, The Ohio State University/OARDC, Wooster, OH, 44691, USA

    Andrea K. Knox & Eric J. Stockinger

  2. Department of Plant Sciences, University of California, Davis, CA, 95616, USA

    Chengxia Li & Jorge Dubcovsky

  3. Agricultural Research Institute of the Hungarian Academy of Sciences, 2462, Martonvasar, Hungary

    Attila Vágújfalvi & Gabor Galiba

  4. Faculty of Information Technology, University of Pannonia, Veszprem, Hungary

    Gabor Galiba

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  1. Andrea K. Knox
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  2. Chengxia Li
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  3. Attila Vágújfalvi
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  4. Gabor Galiba
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  5. Eric J. Stockinger
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  6. Jorge Dubcovsky
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Correspondence to Jorge Dubcovsky.

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Andrea K. Knox, Chengxia Li and Attila Vágújfalvi contributed equally to this work.

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Knox, A.K., Li, C., Vágújfalvi, A. et al. Identification of candidate CBF genes for the frost tolerance locus Fr-A m 2 in Triticum monococcum . Plant Mol Biol 67, 257–270 (2008). https://doi.org/10.1007/s11103-008-9316-6

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  • DOI: https://doi.org/10.1007/s11103-008-9316-6

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