Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 31824
Using Morphological and Microsatellite (SSR) Markers to Assess the Genetic Diversity in Alfalfa (Medicago sativa L.)

Authors: T. Cholastova, D. Knotova


Utilization of diverse germplasm is needed to enhance the genetic diversity of cultivars. The objective of this study was to evaluate the genetic relationships of 98 alfalfa germplasm accessions using morphological traits and SSR markers. From the 98 tested populations, 81 were locals originating in Europe, 17 were introduced from USA, Australia, New Zealand and Canada. Three primers generated 67 polymorphic bands. The average polymorphic information content (PIC) was very high (> 0.90) over all three used primer combinations. Cluster analysis using Unweighted Pair Group Method with Arithmetic Means (UPGMA) and Jaccard´s coefficient grouped the accessions into 2 major clusters with 4 sub-clusters with no correlation between genetic and morphological diversity. The SSR analysis clearly indicated that even with three polymorphic primers, reliable estimation of genetic diversity could be obtained.

Keywords: genetic diversity, Medicago sativa L., morphological traits, SSR markers

Digital Object Identifier (DOI):

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2899


[1] E. Small and M. Jomphe, "A synopsis of the genus Medicago (Leguminosae)," Can. J. Bot., vol. 67, pp. 3260-3294, 1989.
[2] T. J. McCoy and E. T. Bingham, "Cytology and cytogenetics of alfalfa," in Alfalfa and alfalfa improvement. Agronomy Monograph 29 (ed. A. A. Hanson, D. K. Bbarnes and R. R. Hill), Madison USA, 1988, pp. 737- 776.
[3] M. Keivani, S. Sanaz Ramezanpour, H. Soltanloo, R. Choukan, M. Naghavi and M. Ranjbar, "Genetic diversity assessment of alfalfa (Medicago sativa L.) populations using AFLP markers," Australian Journal of Crop Science," vol. 4, no. 7, pp. 491-497, 2010.
[4] A. Mengoni, A. Gori and M. Bazzicalupo, "Use of RAPD and microsatellite (SSR) variation to assess genetic relationships among populations of tetraploid alfalfa, Medicago sativa," Plant Breed., 2000, vol. 119, pp. 311-317.
[5] X. Wang, X. Yang, L. Chen, G. Feng, J. Zhang and L. Jin, "Genetic diversity among alfalfa (Medicago sativa L.) cultivars in Northwest China," Acta Agriculturae Scandinavica, vol. 61, no. 1, pp. 60-66, February 2011
[6] J. R. Xavier, J. Kumar and R. B. Srivastava, "Characterization of genetic structure of alfalfa (Medicago sp.) from trans-Himalaya using RAPD and ISSR markers," African Journal of Biotechnology, vol. 10, no.42, pp. 8176-8187, August 2011.
[7] N. Diwan, A. A.Bhagwat, G. R. Bauchan and P. B. Cregan, "Simple sequence repeat DNA markers in alfalfa and perennial and annual Medicago species," Genomes, vol. 40, pp. 887-895, 1997.
[8] N. Diwan, J. H. Bouton, G. Kochert and P. B. Cregan, "Mapping of simple sequence repeat (SSR) DNA markers in diploid and tetraploid alfalfa," Theor. Appl. Genet., vol. 101, pp. 165-172, 2000.
[9] Z. P. Liu, G. S. Liu and Q. C Yang, "A novel statistical method for assessing SSR variation in autotetraploid alfalfa (Medicago sativa L.)," Genetics and Molecular Biology, 2007, vol. 30, no. 2, pp. 385-391.
[10] V. Vacek, V. Mrázková, A. Sestrienka, J. Sehnalová, I. Bareš and D. Hájek, "Klasifikátor, genus Medicago L., Prague, 1985, pp. 40.
[11] L. Touil, F. Guesmi, K. Fares, C. Zagrouba and A. Ferchichi, "Genetic diversity of some mediterranean populations of the cultivated alfalfa (Medicago sativa L.) using SSR markers," Pakistan Journal of Biological Sciences, vol. 11, no. 15, pp. 1923-1928, 2008.
[12] B. Julier, S. Flajoulot, P. Barre, G. Cardinet, S. Santoni, T. Huguet and Ch. Huyghe, "Construction of two genetic linkage maps in cultivated tetraploid alfalfa (Medicago sativa L.) using microsatellite and AFLP markers," BMC Plant Biology, vol. 3, no. 9, 2003.
[13] S. Kemp, PIC Calculator Extra free software, 2002, (
[14] P. Jaccard, "Étude comparative de la distribuition florale dans une portion des Alpes et des Jura," Bull. Soc. Vandoise Sci. Nat., 1901, vol. 37, pp. 547-579.
[15] FreeTree software v. A. Pavlíček, S. Hrdá and J. Flegr, "FreeTree - freeware program for construction of phylogenic trees on the basis of distance data and bootstrap/jackknife analysis of the tree robustness. Application in the RAPD analysis of the genus Frankelia," Folia Biol., vol. 45, pp. 97-99, 1999. (
[16] STATISTICA version, Copyright StatSoft.Inc., 2005.
[computer program]. version 2012.3.01, Copyright AddinSoft. Inc., 1995 - 2012.
[18] M. V. Bagavathiannan, B. Julier, P. Barre, R. H. Gulden and R. C. Van Acker, "Genetic diversity of feral alfalfa (Medicago sativa L.) populations occurring in Manitoba, Canada and comparison with alfalfa cultivars: an analysis using SSR markers and phenotypic traits," Euphytica, vol. 173, pp. 419-432, 2010.
[19] M. Falahati - Anbaran, A. A. Habashi, M. Esfahany S. A. Mohammadi and B. Ghareyazie, "Population genetic structure based on SSR markers in alfalfa (Medicago sativa L.) from various regions contiguous to the centres of origin of the species," Journal of Genetics, 1997, vol. 86, no. 1, pp. 59-63.
[20] M. L. Crochemore, C. Huyghe, C. Ecalle and B. Julier, "Structuration of alfalfa genetic diversity using agronomic and morphological characteristics. Relationship with RAPD markers," Agronomie, 1998, vol. 18, pp. 79-94.
[21] S. L. Greene, M. Gritsenko and G. Vandermark, "Relating morphological and RAPD marker variation to collection site environment in wild populations of red clover (Trifolium pretense L.)," Genet. Resour. Crop Evol., 2004, vol. 51, pp. 643-653.
[22] X. Zhang, Y. J. Zhang, R. Yan, J. G. Han, H. Fuzeng, J. H. Wang and K. Cao, "Genetic variation of white clover (Trifolium repens L.) collections from China detected by morphological traits, RAPD and SSR," Afr. J. Biotechnol., 2010, vol. 9, no. 21, 3032-3041.
[23] L. Martinez, P. Cavagnaro, R. Masuelli and J. Rodriguez, "Evaluation of diversity of Argentine grapevine (Vitis vinefera L.) varieties using morphological data and AFLP markers," Electron. J. Biotechnol., 2003, vol. 6, pp. 241-250.