Evaluation of genetic diversity in Agropyron cristatum using ISSR molecular marker

Paper Details

Research Paper 01/05/2015
Views (167) Download (4)

Evaluation of genetic diversity in Agropyron cristatum using ISSR molecular marker

Mandana Moradi, Ezatollah Farshadfar, Hooshmand Safari
J. Bio. Env. Sci.6( 5), 425-432, May 2015.
Certificate: JBES 2015 [Generate Certificate]


In order to evaluate genetic diversity, 13 accessions of Agropyron Cristatum were investigated using ISSR molecular markers. The 12 ISSR primers amplified a total of 65 bands of which 60 bands showed polymorphism and 5 bands were monomorphic. Maximum number of bands (8) was related to the primers IS9 and IS13, while minimum bands (4) belonged to the primers IS3 and IS12. The percentage of polymorphic bands (PPB) ranged between 60 and 100. Mean numbers of scored bands (NSB) and polymorphic bands (NPB) per primer were 5.42 and 5, respectively. The PIC values for varied from 0.23 to 0.47 with an average of 0.35. The lowest and the highest PIC indices were recorded for primers IS3 and IS6, respectively. Therefore primer IS6 determined genetic distance much better than other primers so it can be used for the analysis of genetic diversity in agropyron in the future investigations. The primers IS3 with the lowest PIC didn’t have good ability to discriminate genotypes. Cluster analysis based on Dice coefficient of genetic distance classified the accessions into four groups. The first group consisted of genotypes G7 and G13 with average similarity coefficient 0.68. The second group included the accessions 4, G5, G10 and G11 with similarity coefficient 0.71. Genotypes G6, G8 and G9 were located in the third group with similarity coefficient 0.63 The fourth group had the genotypes G1, G2, G3 and G12 with similarity coefficient 0.58, therefore maximum similarity belonged to group 2, while minimum similarity was attributed to group 4. Biplot analysis divided the genotypes into 4 groups which is in accordance with results of cluster analysis. Molecular analysis of variance confirmed significant difference between groupings of accessions based on cluster analysis.


Anderson JA, Church JE, Autrique SD, Thanksley S, Sorrells ME. 1993. Optimizing parental selection for genetic linkage map. Journal of Genome 36(1), 181-188.

Arghavani A, Asghari A, Shokrpour M, Mohammaddost C. 2010. Genetic diversity in ecotypes of two Agropyron Species using RAPD Markers. Research Journal of Environmental Sciences 4, 50-56.

Bor NL. 1970. Gramineae. In: Flora Iranica, Rechinger, K. H. (Ed.). Vol. 70, Akademische Druk-u. Verlagsanstalt, Graz, Austria, 571-573 p.

Che YH, Yang YP, Yang XM, Li XQ, Li LH. 2011. Genetic diversity between ex situ and in situ samples of Agropyron cristatum (L.) Gaertn. Based on simple sequence repeat molecular markers. Crop and Pasture Science 62(8), 639-644.

Chowdhury MA, Vandenberg V, Warkentin T. 2002. Cultivar identification and genetic relationship among selected breeding lines and cultivars in chickpea (Cicer arietinum L.). Euphytica 127, 317– 325.

Chowdhury  RMVK,  Kundu  SJS, Jain  RK. 2008. Applicability of ISSR markers for genetic diversity evaluation in Indian bread wheat genotypes of known origin. Environmental Ecology 26, 126-131.

El-Assal SED, Gaber A. 2012. Discrimination capacity of RAPD, ISSR AND SSR markers and their effectiveness in establishing genetic relationship and diversity among Egyptian and Saudi wheat cultivars. American Journal of Applied Sciences 9(5), 724-735.

Fang DQ, Roose ML. 1997. Identification of closely related citrus cultivars with inter-simple sequence repeats markers. Theoretical and Applied.Genetic 95, 408-417.

Farshadfar M, Farshadfar E. 2004. Evaluation of genetic diversity in Agropyron based on morphological and chemical indices. Agricultural and Natural resource. journal of Science and Technology 8, 243-250.

Ijaz S, Khan I. 2009. Molecular characterization of wheat germplasm using microsatellite markers.Genetic and Molecular Research 8, 809-815.

Iruela M, Rubio J, Cubero JI, Gil J, Millan T. 2002. Phylogenetic analysis in the genus Cicer and cultivated chickpea using RAPD and ISSR markers. Theoretical and Applied Genetic 104, 643–651.

Kuleung C, Baenziger PS, Kachman SD, and Dweikat I. 2006. Evaluating the Genetic Diversity of Triticale with Wheat and Rye SSR Markers. Crop Science 46, 1692.

Manifesto MM, Schlatter A, Hopp HE, Suarez EY, Dubcovsky J. 2001. Quantitative evaluation of genetic diversity in wheat germplasm using molecular markers. Crop Science 41, 682-690.

Najaphy A, Parchin RA, Farshadfar E. 2012. Comparison of phenotypic and molecular characterizations of some important wheat cultivars and advanced breeding lines. Australian Journal of Crop Science 6(2), 326- 332.

Pasqualone A, Lotti C, Bruno A, Vita P, Fonzo N, Blanco A. 2000. Use of ISSR marker for cultivar identification in durum wheat. Options Méditerranéennes: Serie A. Seminaires Méditerranéennes 40, 157-161.

Peakall R, Smouse PE. 2006. Genalex 6: Genetic Analysis in Excel. Population genetic software for teaching and research. Molecular Ecology Notes 6, 288-295.

Powell W, Morgante M, Andre C, Hanafey M, Vogel J, Tingey S, Rafalsky A. 1996. Molecular Breeding 2, 225-238.

Rao NK. 2004. Plant genetic resources: Advancing conservation and use through biotechnology. African Journal of Biotechnology 3(2), 136-145.

Refoufi A, Esnault MA. 2008. Population genetic diversity in the polyploid complex of wheatgrasses using isoenzyme and RAPD data. Biologia Plantarum 52(3), 543-547.

Sofalian O, Chaparzadeh N, Dolati M. 2009. Genetic diversity in spring wheat landraces from northwest of Iran assessed by ISSR markers. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 37(2), 252-256.

Španić V, Buerstmayr H, Drezner G. 2012. Assessment of genetic diversity of wheat genotypes using microsatellite markers. Periodicum biologorum 114, 37-42.

Sudupak A, Akkaya S, Kence A. 2002. Analysis of genetic relationships among perennial and annual Cicer species growing in Turkey using RAPD markers. Theoretical and Applied Genetic 105, 1220–1228.

Szczepaniak M, Bieniek W, Boroń P, Szklarczyk M, Mizianty M. 2009. A contribution to characterisation of genetic variation in some natural Polish populations of Elymus repens (L.) Gould and Elymus hispidus (Opiz) Melderis (Poaceae) as revealed by RAPD markers. Plant Biology 11, 766–773.

Xu J, Conner RL. 1994. Intravarietal variation in satellites and C-banded chromosomes of Agropyron intermedium ssp. trichophorum cv. Greenleaf. Genome 37(2), 305-310.

Xu GH, Su WY, Shu YJ, Cong WW, Wu L, Guo CH. 2012. RAPD and ISSR-assisted identification and development of three new SCAR markers specific for the Thinopyrum elongatum E (Poaceae) genome. Genetic and Molecular Research 11(2), 1741-1751