Comparison of Tetraploid Alfalfa (Medicago sativa L.) Populations Collected from Turkey and Former Soviet Countries

Abstract

Tetraploid Medicago sativa L. subspecies; M. sativa subsp. sativa, M. sativa subsp. falcata and M. sativa subsp. varia form the primary gene pool of the alfalfa. The center of diversity for this subspecies is seen as the Caucasus, Northwest Iran and Northeast Turkey, and its natural range is the former Soviet Union and the southern border of North Africa as the northern border. Genetic diversity among the primary gene pool and comparisons between regions of diversity provides a good reference for breeders when utilizing genetic resources. The United States Department of Agriculture Genetic Resources Information Network (USDA-GRIN) System provides reference data from its entire natural range, including Turkey and the Former Soviet region. In this study, seven populations collected from Turkey and thirteen alfalfa populations collected from Former Soviet Countries held in the USDA GRIN System were evaluated using 20 SSR markers. Within the scope of the study, the information between the locations of the subspecies was compared to reveal the hierarchical population structure. The results obtained from the STRUCTURE and PCA analyzes show that the populations are clustered in two main groups for both countries, but there is a high similarity in hybrid genome contents in the subspecies belonging to the Former Soviet countries. In addition, it was determined by AMOVA analysis that the variance within the populations was higher than that between the populations according to the subspecies analyzed from both countries. It is thought that the results will be effective in terms of using alfalfa genetic resources of these countries in breeding programs.

Keywords

• Tetraploid alfalfa
• Turkey
• Former Soviet
• Population
• SSR markers

References

1. Brummer, E.C., Kochert, G., & Bouton, J.H. (1991). RFLP variation in diploid and tetraploid alfalfa. Theor. Appl. Genet. 83:89–96.
2. Doyle, J.J., & Doyle, J.L. (1990). Isolation of plant DNA from fresh tissue. Focus 12:13–15.
3. Diwan, N., Bouton. J.H., Kochert, G., Cregan, P.B. (2000). Mapping of simple sequnce repeat (SSR) DNA markers in diploit and tetraploit alfalfa. Theor Appl Genet., 101: 165-172.
4. Eren, B., Keskin, B., Demirel, F. et al. (2022). Assessment of genetic diversity and population structure in local alfalfa genotypes using iPBS molecular markers. Genet Resour Crop Evol. https://doi.org/10.1007/s10722-022-01450-2
5. Evanno, G., Regnaut, S., & Goudet, J. (2005). Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620.
6. Hanson., A.A., Barnes, D.K., & Hill, J.R. (1988). ‘’Alfalfa and Alfalfa Improvement’’, ISBN: 0-89118-094-X, American Society of Agronomy, Inc., Crop Science Society of America, Inc., Soil Science Society of America, Inc., Madison, Wisconsin, USA.
7. Havananda, T., Brummer, E.C., Maureira-Butler, I.J., Doyle, J.J. (2010). Relationships among Diploit Members of the Medicago sativa (Fabaceae) Species Complex Based on Chloroplast and Mitochondrial DNA Sequences. Systematic Botany. 35(1): 140-150.
8. Ilhan, D., Li, X., Brummer, E.C., Sakiroglu, M. (2016). Genetic Diversity and Population Structure of Tetraploid Accessions of the Medicago sativa-falcata Complex. Crop Sci. 56:1-11.

9. Johnson, L.B., & Palmer, J.D. (1989). Heteroplasmy of chloroplast DNA in Medicago. Plant Molecular Biology. 12:3-11.
10. Julier, B., Flajoulot, S., Barre, P., Cardinet, G., Santoni, S., Huguet, T., Huyghe, C. (2003). Construction of two genetic linkage maps in cultivated tetraploit alfalfa (Medicago sativa) using microsatellite and AFLP markers. BMC Plant Biol., 3:9.
11. Kalo, P., Endre, G., Zimanyi, L., Csanadi, G., Kiss, G.B. (2000). Construction of an improved linkage map of diploid alfalfa (Medicago sativa). Theor. Appl. Genet. 100 (5): 641-657.
12. Mengoni, A., Gori, A., Bazzicalupo, M. (2000). Use of RAPD and Microsatellite (SSR) Variation to Assess Genetic Relationships among Populations of Tetraploit Alfalfa, Medicago sativa. Plant Breed. 119:311-317.
13. Michaud, R., Lehman, W.F., & Rumbaugh, M.D. (1988). World distribution and historical development. In A.A. Hanson[ed.], D.K. Barnes and R.R. Hill Jr. [co-eds], Alfalfa and alfalfa improvement, Agronomy monograph 29, 93-124. American Society of Agronomy, Crop Science Society of Agronomy, and Soil Science Society of Agronomy, Madison, Wisconsin, USA.
14. Muller, M-H., Prosperi, J.M., Santoni, S., Ronfort, J. (2003). Inferences from mitochondrial DNA patterns on the domestication history of Alfalfa (Medicago sativa). Molecular Ecology, 12:2187-2199.
15. Peakall, R., & Smouse, P.E. (2001). GenAlEx V6: genetic analysis in excel population genetic software for teaching and research. Australian National University, Canberra. http://wwwanueduau/BoZo/ GenAlE.
16. Pritchard, J.K., Stevens, M., & Donnelly, P. (2000). Inference of population structure using multilocus genotype data. Genetics 155:945–959.
17. Quiros, C.F., & Bauchan, G.R. (1988). The Genus Medicago and the Origin of the Medicago sativa Complex, 93-124.
18. Robins, J.G., Luth, D., Campbell, T.A., Bauchan, G.R., He, C., Viands, D.R., Hansen, J.L., Brummer, E.C. (2007). Genetic mapping of biomass production in tetraploid alfalfa (Medicago sativa L.) Crop Sci 47:1-10
19. Schuelke, M. (2000). An economic method for the fluorescent labeling of PCR fragments. Nat Biotechnol 18:233–234.
20. Schumann, C.M., & Hancock, J.F. (1989). Paternal inheritance of plastids in Medicago sativa. Theor. Appl. Genet. 78:863-866.
21. Segovia-Lerma, A., Cantrell, R.G., Conway, J.M., & Ray, I.M. (2003). AFLP-based assessment of genetic diversity among nine alfalfa germplasms using bulk DNA templates. Genome 46:51– 58.
22. Sledge, M.K., Ray, I.M., Jiang, G. (2005). An expressed sequence tag SSR map of tetraploid alfalfa (Medicago sativa L.) Theor Appl Genet, 111: 980-992.
23. Sakiroglu, M., Doyle, J.J., & Brummer, E.C. (2010). Inferring population structure and genetic diversity of broad range of wild diploid alfalfa (Medicago sativa L.) accessions using SSR Markers. Theor. Appl. Genet. 121:403–415.
24. Şakiroğlu, M., İlhan, D., Kaya, M.M., Demirözoğul, O., Uluçay, O., & Eren, B. (2011). Moleküler Veriler Işığında Medicago sativa L. Tür Kompleksinin Mevcut Durumu. Kafkas Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 4(1), 32-42.
25. Sakiroglu, M., & Kaya, M.M. (2012). Estimating genome size and confirming ploidy levels of wild tetraploid alfalfa accessions (Medicago sativa subsp. varia) using flow cytometry. Turk. J. Field Crop 17:151–156.
26. Şakiroğlu, M., Eren, B., İlhan, D., & Tufan, T. (2015). Historical alfalfa landraces perform higher yield under dry farming in Turkey. Procedia Environmental Sciences, 29, 189.
27. Tanksley, S.D., & McCouch, S.R. (1997). Seed Banks and Molecular Maps: Unlocking Genetic Potential from the Wild. Science, 277 (5329): 1063-1066.
28. Vyšniauskiene, R., Naugžemys, D., Patamsyte, J., Rancˇeliene, V., Cesniene, T., & Žvingilva, D. (2015). ISSR and chloroplast DNA analyses indicate frequent hybridization of alien Medicago sativa subsp. sativa and native M. sativa subsp. falcata. Plant Syst. Evol. 301:2341–2350.
29. Yu, K., & Pauls, K.P. (1993). Rapid estimation of genetic relatedness among heterogeneous populations of alfalfa by random amplification of bulked genomic DNA samples. Theor. Appl. Genet. 86:788–794.