Study of genetic diversity among 17 populations of alfalfa (Medicago sativa) using cytogenetic and ISSR molecular markers

Document Type : Research Paper

Authors

Abstract

Alfalfa (Medicago sativa L.) is one of the most important forage crops because of high quality, growing extensively in farms and rangelands of Iran. Genetic variability is one of the most aspects for crop improvement. The experiments were carried out to determine genetic variability among 17 alfalfa populations based on ISSR and cytogenetic markers. All of the tested ISSR primers showed 44 visible bands. Three primers had the most number of bands while another primers showed the least band numbers. Mean percentage of polymorphism in the primers was 90.18 percent. Polymorphic information content (PIC) and marker index (MI) average were 0.36 and 1.88 respectively. Cluster analysis classified the populations genotype into four clusters. In order to investigate karyotipic variation between the populations, the populations were studied for several karyotipic characteristics. Including long arm, short arm and total length of the chromosomes, long arm to short arm and short arm to long arm ratios and total form percentage (TF%). All of the genotypes had 32 chromosomes. Analysis of variance of mitotic data in completely randomized design, showed highly significant differences between the studied genotypes for all of the studied traits (p<0.01). According to the Stebbins classification all of the genotypes located in A1 class. Cluster analyses grouped the genotypes into four clusters.

Keywords


-      Azhar Hossain, M. and Bauchan, G.R., 2001. Distribution and characterization of heterochromatic DNA in the tetraploid African populations of alfalfa Genome. Crop Science, 41: 1921-1926.
-      Bahar, M., Ghobadi, S., Erfani Moghadam, V., Pamchi, A., Talebibadaf, M., Kaboli, M. and Mokhtarzade, A., 2006. Evaluation of genetic diversity of local lucerne population using ESTS markers. Journal of Science and Technology in Agriculture and Natural Resources, 2: 142-153.
-      Bauchan, G.R., Small, E., Brooks, B. and Auricht, G.C., 1995. A systematic comparison of early-and late-flowering forms of Medicago scutellata. Canadian Journal of Botany, 71:183-192.
-      Bauchan, G.R. and Azhar Hossain, M., 1998. Karyotypic analysis of C-banded chromosomes of diploid alfalfa: Medicago sativa ssp. caerulea and ssp. falcata and their hybrid. Journal of Heredity, 89:191-193.
-      Bolourchian, F., Farshadfar, M., Safari, H. and Shirvani, H., 2013. Evaluation of genetic diversity in different genotypes of (Medicago Sativa L.) using ISSR markers. International Journal of Farming and Allied Sciences, 2: 995-960.
-      Farshadfar, M., Kissimon, T. and Sutka, T., 1995. Genetic distance between Triticum timopheery zhuk, T. araraticum takubz. Plant Breeding, 114: 401-405.
-      Ghanavati, F., Mozafari, J. and Kazempour, Sh., 2006. Familiar relationship between lucerne genuse on Iran based on RAPD molecular marker. Pajouhesh & Sazandegi, 66: 2-12 (In Persian).
-     Gupta, P.K., 1995. Cytogenetic. Rastogiand Company. Maruat India, 3-4.
-      Hamza, S., Hamida, W.B., Rebai, A. and Hrrabi, M., 2004. SSR-based genetic diversity assessment among Tunisian winter barley and relationship with morphological traits. Euphytica, 135:107-118.
-      Hou, Y., Yan, Z., Wei, Y., 2005. Genetic diversity in barely from west China based on RAPD and ISSR nalysis. Barely Genetics Newsletter, 35:9-22.
-      Huziwara, Y., 1962. Karyotype analysis in some genera of Composite. VIII. Further studies on the chromosome of aster. American Journal of Botany, 49: 116-119.
-      Levan, A., Fedga, K. and Sandberg, A.A., 1964. Nomenclature for centromeric position on chromosomes. Hereditas, 52: 201-220.
-      Majd, A. and Sharyatzade, M.A., 2001. Molecular Biology, Aeyj publisher, Teharan, 731pp (In Persian).
-      Mantel, N., 1967. The detection of disease clustering and a generalized regression approach. Cancer Research, 27: 209-220.
-      Mengoni, A., Gori, A. and Bazzicalupo, M., 2000. Use of RAPD and microsatellite (SSR) variation to assess genetic relationships among populations of tetraploid alfalfa, Meddicago sativa. Plant Breeding, 119:311-317.
-     Michaud, R., Tremblay, G.F., Belanger, G. and Michaud, M., 1990. Proteolysis in alfalfa silages made from different cultivars. Journal Agriculture and Agri-Food Canada, Soils and Crops Research and Development Center, 2560 Hochelaga blvd, Sainte-Foy, Quebec, Canada, G1V 213.
-      Michaud, R., Tremblay, G.F., Belanger, G. and Michaud, J., 1991. Agriculture and Agri-Food Canada, Soils and Crops Research and Development Center, 2560 Hochelaga blvd, Sainte-Foy, Quebec, Canada, G1V 213.
-      Mirzaie-Nodoushan, H., Mehrpur, Sh., Rezaie, M. and Rashvand, S., 2002. Primary karyotypic investigation on several populations of Aloe litoralis. Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 9: 49-84, (In Persian).
-      Mirzaie -Nodoushan, H., Mehrpur, Sh. and Sifidkon, F., 2006. Path analysis of the characters influencing essential oil in three Thymus species. Pajuohesh & Sazandegi, 70:88-94 (In Persian).
-      Mortazavi, M., 2007. Study of genetic variability of lucerne based on morphological, cytogenetic and biochemeical markers and evaluation of drought tolerance. M.Sc. thesis, Razi University, Kermanshah, (In Persian).
-      Powell, W., Morgante, M., Andre, C., Hanafey, M., Vogel, J., Tingey, S. and Rafalski, A., 1996. The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Molecular Breeding, 2: 225-238.
-      Reddy, M.P., Sarla, N. and Siddiq, E.A., 2002. Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica, 128:9-17.
-      Rezaei, A., 2001. Lucerne Breeding. Centre of University Publishing. Tehran, 233pp, (In Persian).
-      Romero-Zarco, C., 1986. A new method for estimating karyotype asymmetry. Taxonomy, 35: 526-530.
-      Ronald, L. and Indra, K., 2001. NA-Based Markers in Plants. Kluwer Academic Publisher.
-      Schaut, J.W., Qi, X. and Stam, P., 1997. Association between relationship measures based on AFLP markers, pedigree data and morphological traits in barley. Theoretical and Applied Genetics, 95: 1161-1168.
-      Shahnejat Bousheri, A., Fatahi, H.A., Yazdi Samadi, B., 2008. Study of genetic variation among varieties and population of lucerne based on RAPD markers. Modern Genetics, 3: 35-43 (In Persian).
-      Sharyat, A., 2001. Study of genetic variability of annual lucerne by cytogenetic, electrophoretic and morphological traits. M.Sc. thesis, Sistan and Balouchestan University (In Persian).
-      Sicard, D., Nanni, L., Porfiri, O., Bulfon, D., and Papa, R., 2005. Genetic diversity of Phaseolus vulgaris L. and P. coccineus L. landraces in central Italy. Plant Breeding, 124: 464–472.
-      Stebbins, G.L., 1971. Chromosome Evolution in Higher Plants. Edward Arnold Publisher. LTD. London, pp.216.
-      Terzopoulos, P.J. and Bebeli, P.J., 2008. Genetic diversity analysis of Mediterranean faba bean (Vicia faba L.) with ISSR markers. Field Crops Research, 108:39-44.
-      Torres, A.M., Weeden, N.F. and Martin, A., 1993. Linkage among isozyme, RFLP and RAPD markers in Vicia faba. Theoretical and Applied Genetics, 85: 935–945.
-      Tucak, M., Popovic, S., Cupic, T., Grljusic, S., Bolaric, S., and Kozumplik, V., 2008. Genetic diversity of alfalfa (Medicago spp.) estimated by molecular markers and morphological characters. Periodicum Biologorum, 110: 243-249.
Vern, L.M. and Donald, L., 2006. Testing alfalfa for its feeding value. Division of Agriculture and Natural Resources. University of California. Rep, 109.