Abstract
In alfalfa (Medicago sativa), an autotetraploid forage legume, stem length is a major component of forage yield, quality and competing ability. In this species, flowering date is not a breeding criterion. Association mapping based on a candidate gene approach has given good results in plants, including autotetraploid species for which genetic analyses are complex. The role of a CONSTANS-LIKE gene, identified as a candidate for stem elongation and flowering date in the model legume M. truncatula, was tested for association with the same traits in alfalfa. Four hundred genotypes from ten cultivars were evaluated for stem height and flowering date in two locations during 4 years. They were genotyped with simple sequence repeat markers and a low structuration was noticed. Primers were designed to amplify and sequence two regions of the alfalfa gene homologous to CONSTANS-LIKE. Single nucleotide polymorphisms (SNPs) were detected and their allelic dose in each genotype was scored. Linkage disequilibrium within CONSTANS-LIKE rapidly decreased as expected. Eight SNPs with a frequency above 10% were detected over 1,010 bp (one SNP every 126 bp on average) in the 400 genotypes. This number was lower than observed in a neutral gene (a SNP every 31 bp on average). Highly significant associations of three SNPs to flowering date and stem height were identified. Each SNP explained up to 4.2% of the genetic variance. Thus, as in the model species, the CONSTANS-LIKE gene was shown to be involved in flowering date and stem height in alfalfa.
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Acknowledgments
The authors thank C. Gibelin, A. Weber, J.F. Bourcier, A. Faure and J. Jousse for their technical assistance in the lab and field. This work was part of a contract between ACVF (Association des Créateurs de Variétés Fourragères) and INRA, and was supported by the Ministry of Agriculture and Fisheries (Contrat de Branches 2005–2008). D. Herrmann received a grant from the Plant Breeding Division of INRA.
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Communicated by T. Luebberstedt.
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Fig. S1 Example for the estimation of allele dose (SNP e332) for the five genotypic classes. F, one of the four alleles has a deletion of 1 bp; G, one allele has a deletion of 3 bp (DOC 106 kb)
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Herrmann, D., Barre, P., Santoni, S. et al. Association of a CONSTANS-LIKE gene to flowering and height in autotetraploid alfalfa. Theor Appl Genet 121, 865–876 (2010). https://doi.org/10.1007/s00122-010-1356-z
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DOI: https://doi.org/10.1007/s00122-010-1356-z