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Theoretical and Applied Genetics

, Volume 126, Issue 7, pp 1885–1896 | Cite as

Cloning and characterization of chromosomal markers in alfalfa (Medicago sativa L.)

  • Feng Yu
  • Yunting Lei
  • Yuan Li
  • Quanwen Dou
  • Haiqing Wang
  • Zhiguo Chen
Original Paper

Abstract

Eleven tandemly repetitive sequences were identified from a Cot-1 library by FISH and sequence analysis of alfalfa (Medicago sativa). Five repetitive sequences (MsCR-1, MsCR-2, MsCR-3, MsCR-4, and MsCR-5) were centromeric or pericentromeric, of which three were satellite DNAs and two were minisatellite DNAs. Monomers of 144, 148, and 168 bp were identified in MsCR-1, MsCR-2, and MsCR-3, respectively, while 15 and 39 bp monomers were identified in MsCR-4 and MsCR-5, respectively. Three repetitive sequences were characterized as subtelomeric; one repetitive sequence, MsTR-1, had a 184 bp monomer, and two repetitive sequences had fragments of 204 and 327 bp. Sequence analysis revealed homology (70–80 %) between MsTR-1 and a highly repeated sequence (C300) isolated from M. ssp. caerulea. Three identified repetitive sequences produced hybridization signals at multiple sites in a few of the chromosomes; one repetitive sequence was identified as the E180 satellite DNA previously isolated from M. sativa, while the other 163 and 227 bp fragments had distinct sequences. Physical mapping of the repetitive sequences with double-target FISH revealed different patterns. Thus, nine novel tandemly repetitive sequences that can be adopted as distinct chromosome markers in alfalfa were identified in this study. Furthermore, the chromosome distribution of each sequence was well described. Though significant chromosome variations were detected within and between cultivars, a molecular karyotype of alfalfa was suggested with the chromosome markers we identified. Therefore, these novel chromosome markers will still be a powerful tool for genome composition analysis, phylogenetic studies, and breeding applications.

Keywords

Repetitive Sequence Chromosome Marker Strong Hybridization Signal E180 Signal Minisatellite DNAs 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We thank Professor Tao Wang (China Agriculture University) for providing the seeds of some alfalfa materials. This study was financially supported by the “Joint Scholars” program of the “Lights in the Western Region” talent cultivation plan of the Chinese Academy of Sciences and partly supported by the Main Direction Program for Knowledge Innovation of the Chinese Academy of Sciences (KSCX2-EW-Q-23).

Supplementary material

122_2013_2103_MOESM1_ESM.doc (119 kb)
Supplementary material 1 (DOC 119 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Feng Yu
    • 1
    • 2
  • Yunting Lei
    • 1
  • Yuan Li
    • 1
    • 2
  • Quanwen Dou
    • 1
  • Haiqing Wang
    • 1
  • Zhiguo Chen
    • 1
  1. 1.Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Plateau Institute of BiologyChinese Academy of SciencesXiningChina
  2. 2.Graduate University of Chinese Academy of SciencesBeijingChina

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