Iranian Journal of Field Crop Science

Iranian Journal of Field Crop Science

Analysis of genetic and cytogenetic variations between alfalfa (Medicago sativa L.) genotypes in Iran

Document Type : Research Paper

Authors
Mohsen Farshadfar 1
Fatemeh Boloorchian 2
Hooshmand Safari 3
Hooman Shirvani 4
1 Associate Professor, Department of Agriculture, Payame Noor University, Iran
2 Former M. Sc. Student, Young Researchers Club, Islamic Azad University, Kermanshah Branch, Kermanshah, Iran
3 Assistant Professor, Research Center of Agriculture & Natural Resources, Organization of Research, Agricultural Education, Kermanshah, Iran
4 Teacher, Department of Agriculture, Payame Noor University, Iran
10.22059/ijfcs.2017.208241.654123
Abstract
Alfalfa (Medicago sativa L.) is one of the important forage crops producing highly nutritious biomass and adaptability. In this study the genetic diversity among 19 genotypes of Medicago sativa was evaluatedbased on the molecular and cytogenetic markers. Molecular studies were carried out based on eight Inter Simple Sequence Repeat (ISSR) primers. Cytogenetic data were obtained by karyotype analysis with mitotic chromosomes. There were significant differences between cytogenetic characterictics (P≤0.01). The genotypes ES-211, ES-027, ES 037, ES-065, and ES-199 had the greatest chromatin content and the highest asymmetrical value as well. The greatest amount of asymmetry index (AI) belonged to ES211, E119 while the least value of AI was ES037. The genotype ES119 had the greatest CVCL, Mean centromeric asymmetry (MCA), r-value and relative length (RL%) characterictics. Analysis of banding patterns of eight ISSR primers revealed 29 polymorphic bands. A total of 29 polymorphic fragments were scored and the average of polymorphic information content (PIC) was 0.32. The number of polymorphic fragments for each primer varied from three to six with mean of 4.62 polymorphic fragments per primer. The mean polymorphic percentage was 86%. The Cluster analysis of molecular data classified all 19 genotypes into three different groups. According to the karyotypic characterictics, all genotypes were grouped to three clusters as well.
Keywords
Genetic diversity
karyotype
ISSR
Medicago sativa L
Subjects
  1. Arano, H. & Satio, H. (1980). Cytological studies in family Umbelliferae 5. Karyotypes of seven species in subtribe Seselinae. Kromosomo, 2(17), 471-480.
  2. Azhar Hossain, M. & Bauchan, G. R. (2001). Distribution and characterization of heterochromatic DNA in the tetraploid African populations of alfalfa Genome. Crop Science, 41, 1921-1926.
  3. Azizi, N., Amirouche, R. & Amirouche, N. (2016). Cytotaxonomic Diversity of some Medicinal Species of Hyacinthaceae from Algeria. Pharmacogn, 6(1), 34-38.
  4. Barcaccia, G., Albertin, E., Tavoletti, S., Falcinelli, M. & Veronesi, F. (1999). AFLP fingerprinting in Medicago spp: its development and application in linkage mapping. Plant Breeding, 118, 335-341.
  5. Barnes, D. K., Bingham, E., Murphy, T., Hunt, R. P., Beard, O. J., Skrdla, D. F. & Teuber, L. R. (1977). Alfalfa Germplasm in the United States: Genetic Vulnerability, Use, Improvement and Maintenance. U.S. Department of Agriculture, U.S. Printing Office. Technical Bulletin, 1571.
  6. Bauchan, G. R. & 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.
  7. Bauchan, G. R. & Campbell, T. A. (1994). Use of an image analysis system to karyotype alfalfa, Medicago sativa. Heredity, 85, 18-22.
  8. Bauchan, G. R., Small, E., Brooks, B. & Auricht, G. C. (1995). A systematic comparison of early-and late-flowering forms of Medicago scutellata. Canadian Journal of Botany, 71, 183-192.
  9. Bernadette, J., Sandrine, V., Philippe, B., Gaelle, C., Sylvain, S., Thierry, H. & Christian, H. (2003). Construction of two genetic linkage maps in cultivated tetraploid alfalfa (Medicago sativa) using microsatellite and AFLP markers. BMC Plant Biology, 3, 1-19.
  10. Brantestam, A. K., Botheme, R. V., Dayteg, Ch., Rashall, Tuvesson, S. & Weibull, J. (2004). Inter simple sequence repeat analysis of diversity and relationships in cultivated burly of Nordic and Baltic origin. Hereditas, 141, 186-192.
  11. Brummer, E. C. (1999). Capturing hetrosis in forage crop cultivar development. Crop Science, 939- 943.
  12. Ceccarelli, M. S., Minelli, M., Falcinelli, M. & Cionini, P. G. (1994). Genome size and plant development in hexaploid Fustuca arundinacea. Heredity, 71 555-560 Chawla, H.S. 2004. Labroratory Manual for plant Biotechnology.
  13. Diwan, N., Bhagwat, A. A., Bauchan, G. R. & Cregan, P. B. (1997). Simple sequence repeat (SSR) DNA markers in alfalfa and perennial and annual Medicago species. Genome, 40, 887-895.
  14. Doyle, J. J. & Doyle, J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 19, 11-15.
  15. Ellwood, S. E., D’Souza, N. K., Kamphuis, L. G., Burgess, T. I., Nair, R. M. & Oliver, R. P. (2006). SSR analysis of the Medicago truncatula SARDI core collection reveals substantial diversity and unusual genotype dispersal throughout the Mediterranean basin. Theoretical and Applied Genetics, 112, 977-983.
  16. Ghanavati, F., Mozafari, J. & Kazempour, Sh. (2006). Familiar relationship between Lucerne genuse on Iran based on RAPD molecular marker. Pajouhesh & Sazandegi, 66, 2-12. (in Farsi)
  17. Hieter, P. & Griffiths, T. (1999). Polyploidy more is more of less. Science, 285, 210- 211.
  18. 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.
  19. Kumari, G. & Bijoy, K. R. (2010). Karyotype studies in dominant species of Aloe from eastern India. Caryologia, 63(1), 41-49.
  20. Levan, A., Fedga, K. & Sandberg, A. A. (1964). Nomenclature for centromeric position on chromosomes. Hereditas, 52, 201-220.
  21. Li, Y., Wng, Y., Sun, X. & Han, J. (2009). Using microsattelite (SSR) and morphological markers to assess the genetic diversity of 12 alfalfa (Medicago sativa spp. Falcata) populations from Eurasia.  Biotechnology, 8(10), 2102-2108.
  22. Liliya, K. (2000). Organization of plant genetic resources in Bulgaria. Acta Horticulturae, 510, 247-259.
  23. Manifesto, M. M., Schlatter, A.S., Hopp, H.E., Suarez, E. Y. & Dubcovky, J. (2001). Quantitative evaluation of genetic diversity germplasm using molecular markers. Crop Science, 41, 682-690.
  24. Marvyan, S., Safarnejad, A. & Majd, A. (2002). Comparison of some Caryological traits of Lucerne cultivars. Pajouhesh & Sazandegi, 56, 57, 25-29. 
  25. Mortazavi, M. (2007). Evaluation of genetic variation of Lucerne using morphological, cytogenetic and biochemical markers. M.Sc. Thesis, Razi University, Kermanshah, Iran. (in Farsi)
  26. Paknia, R. & Karimzadeh, G. (2011). Karyotypic Study and Chromosome Evolution in Some Iranian Local Onion Populations. Journal of Plant Physiology and Breeding, 1(1), 49-62.
  27. Paszko, B. (2006). A critical review and a new proposal of karyotype asymmetry indices. Plant Systematics and Evolution, 258(1-2), 39-48.
  28. Peng, L. Z., Gong, L. S. & Qingm, C. Y. (2007). A novel statistical method for assessing SSR variation in autotetraploid alfalfa (Medicago sativa L.). Genetics Molecular Biology, 30, 385-391.
  29. Powell, W., Morgante, M., Andre, C., Hanafey, M., Vogel, J., Tingey, S. & Rafalski, A. (1996). The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Molecular Breeding, 2, 225-238.
  30. Price, H. J., Chmber, K. L. & Bachmann, K. (1981).Geographic and ecological distribution of genome DNA content variation in Microseris douglasii (Asteracea). Botanical Gazette, 142, 415-426.
  31. Romero-Zarco, C. (1986). A new method for estimating karyotype asymmetry. Taxonomy, 35, 526-530.
  32. Ronfort, J., Jenczewski, E., Bataillon, T. & Rousset, F. (1998). Analysis of population structure in autotetraploid species. Genetics, 150, 921-930.
  33. Sandrine, F., Joëlle, R., Pierre, B., Philippe, B., Thierry, H., Christian, H. & Bernadette, J. (2008). Genetic diversity among alfalfa (Medicago sativa) cultivars coming from a breeding program, using SSR markers. Theoretical and Applied Genetics, 111, 1420-1429.
  34. Sharyat, A. (2001). Study of genetic variability of annual Lucerne by cytogenetic, electrophoretic and morphological traits. M.Sc. Thesis, Sistan and Balouchestan University, Iran. (in Farsi)
  35. Stebbins, G. L. (1971). Chromosomal Evolution in Higher Plants. Edward Arnold, London, UK.
  36. Surve-Iyer, R. S., Adams, G. C., Lezzon, A. F. & Jones, A. L. (1995). Isozyme detection and variation in Ieucostoma species from prunus and malus. Mycologia, 87, 471- 482.
  37. Veronesi, F. Charles, B. & Huyghe, C. (2010). Alfalfa. Springer Science, 395-436.
  38. Watanabe, K., Yahara, T., Denda, T. & Kosuge, K. (1999). Chromosomal evolution in the genus Brachyscome (Asteraceae, Astereae): Statistical tests regarding correlation between changes in karyotype and habit using phylogenetic information. Journal of Plant Research, 112(2), 145-61.
Iranian Journal of Field Crop Science
Volume 48, Issue 3 - Serial Number 3
Autumn 2017
Pages 695-708

  • Receive Date 22 June 2016
  • Revise Date 26 November 2016
  • Accept Date 03 December 2016
  • Publish Date 22 November 2017