Plant Molecular Biology

, Volume 67, Issue 3, pp 257–270 | Cite as

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

  • Andrea K. Knox
  • Chengxia Li
  • Attila Vágújfalvi
  • Gabor Galiba
  • Eric J. Stockinger
  • Jorge DubcovskyEmail author


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.


Frost tolerance COR14b DHN5 WCS120 CBF Wheat Triticum monococcum 



Base pair


C-Repeat binding factor


Cold regulated


C-Repeat/Dehydration response element




Frost resistance


Quantitative trait locus


Standard errors of the means


Single nucleotide polymorphism


Supplementary online material



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.

Supplementary material

11103_2008_9316_MOESM1_ESM.doc (125 kb)
(DOC 125 kb)


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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Andrea K. Knox
    • 1
  • Chengxia Li
    • 2
  • Attila Vágújfalvi
    • 3
  • Gabor Galiba
    • 3
    • 4
  • Eric J. Stockinger
    • 1
  • Jorge Dubcovsky
    • 2
    Email author
  1. 1.Department of Horticulture and Crop ScienceThe Ohio State University/OARDCWoosterUSA
  2. 2.Department of Plant SciencesUniversity of CaliforniaDavisUSA
  3. 3.Agricultural Research Institute of the Hungarian Academy of SciencesMartonvasarHungary
  4. 4.Faculty of Information TechnologyUniversity of PannoniaVeszpremHungary

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