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Dehydrin variants associated with superior freezing tolerance in alfalfa (Medicago sativa L.)

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Abstract

A cDNA (msaCIG) encoding a cold-inducible Y2K4 dehydrin in alfalfa (Medicago sativa spp. sativa) was shown to share extensive homology with sequences from other species and subspecies of Medicago. Differences were mainly the result of the occurrence of large indels, amino acids substitutions/deletions and sequence duplications. Using a combination of a bulk segregant analysis and RFLP hybridization, we uncovered an msaCIG polymorphism that increases in frequency in response to recurrent selection for superior freezing tolerance. Progenies from crosses between genotypes with (D+) or without (D−) the polymorphic dehydrin significantly differed in their tolerance to subfreezing temperatures. Based on the msaCIG sequence, we looked for intragenic variations that could be associated to the polymorphism detected on Southern blots. Amplifications with primers targeting the 3′ half side of msaCIG revealed fragment size variations between pools of genotypes with (+) or without (−) the polymorphism. Three major groups of amplicons of ≈370 nt (G1), 330 nt (G2), and 290 nt (G3) were distinguished. The G2 group was more intensively amplified in pools of genotypes with the polymorphic dehydrin and was associated to a superior freezing tolerance phenotype. Sequences analysis revealed that size variation in the 3′ half was attributable to the variable occurrence of large indels. Single amino acid substitutions and/or deletions caused major differences in the prediction of the secondary structure of the polypeptides. The identification of dehydrin variants associated to superior freezing tolerance paves the way to the development of functional markers and the fixation of favorable alleles in various genetic backgrounds.

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Acknowledgments

The authors acknowledge the contribution of Mrs. Josée Bourassa, Josée Michaud and Pierre Lechasseur for their technical assistance. They would also like to thank Dr Isobel Parkin from Agriculture and Agri-Food Canada in Saskatoon for her valuable comments on the manuscript. The contribution of Mr David Gagné for bioinformatics analyses is highly appreciated.

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  1. Wilfried Rémus-Borel

    Present address: Laboratoire de Bio-Contrôle, Centre de Recherche en Horticulture, Pavillon de l’Envirotron, Université Laval, Quebec, QC, G1V-0A6, Canada

Authors and Affiliations

  1. Crops and Soils Research and Development Center, Agriculture and Agri-Food Canada, 2560 Hochelaga Blvd., Quebec, QC, G1V-2J3, Canada

    Wilfried Rémus-Borel, Yves Castonguay, Jean Cloutier, Réal Michaud, Annick Bertrand, Réjean Desgagnés & Serge Laberge

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  1. Wilfried Rémus-Borel
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Correspondence to Yves Castonguay.

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Communicated by T. Close.

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Rémus-Borel, W., Castonguay, Y., Cloutier, J. et al. Dehydrin variants associated with superior freezing tolerance in alfalfa (Medicago sativa L.). Theor Appl Genet 120, 1163–1174 (2010). https://doi.org/10.1007/s00122-009-1243-7

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  • DOI: https://doi.org/10.1007/s00122-009-1243-7

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