Pomegranate is one of the most important horticultural crops in Iran, and has been cultivated for thousands of years in this country. At this period due to selection of superior cultivars from nature or mutation emerged in these cultivars, and their vegetative propagation, substantial genetic variation has occurred within and among the cultivars. Thus, each cultivar may consist of different clones. According to this issue, diversity within four commercial cultivars of pomegranate was analyzed. Two molecular marker systems including ISSR and SSR were used to evaluate variability between 36 samples of four commercial cultivars. ISSR markers produced 114 amplification products, out of which 97 were polymorphic (83.23%). Mean resolving power was 2.96 for ISSR markers. 19 SSR molecular markers were used, 15 of which amplified polymorphic products, while the remaining ones monomorphic., The number of polymorphic alleles per locus ranged from two to four (average 3.6). The observed and expected heterozygosities ranged from 0.04 to 0.92 and 0.14 to 0.62, respectively. In addition, mean polymorphic information content was 0.45 for SSR loci. Our results showed that commercial Iranian pomegranate have different clones. Therefore, ISSR and SSR markers can be a useful tools for detecting clones of each cultivar.
___
Ajal EA, Jbir R, Melgarejo P, Hernández F, Haddioui A, & Hannachi AS. 2014. Efficiency of inter simple sequence repeat (ISSR) markers for the assessment of genetic diversity of Moroccan pomegranate (Punica granatum L.) cultivars. Biochem. Syst. Ecol., 56:24–31
Bassam BJ, Caetano-Anolles G. 1993. Silver staining of DNA in polyacrylamide gels. Appl. Bioch. and Biotech., 42:181–188
Behzadi Shahrbabaki H. 1998. Genetic diversity of pomegranate genotypes in Iran. Nashr Amoozesh Keshavarzi pp 265.
Curro S, Caruso M, Distefano G, Gentile A, La Malfa S. 2010. New microsatellite loci for pomegranate, Punica granatum (Lythraceae). Am. J. Bot., 1: 58-60.
Ebrahimi S, Sayed-Tabatabaei BE, Sharifnabi B. 2010. Microsatellite isolation and characterization in pomegranate (Punica granatum L.). Iran. J. Biotechnol., 8: 159–163
Gil MI, Barberan FAT, Pierce BH, Holcroft DM, Kader AA. 2000. Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing. J. Agric. Food. Chem., 48: 4581–89
Gilbert JE, Lewis RV, Wilkinson MJ, Galigari PDS. 1999. Developing and appropriate strategy to assess genetic variability in plant germplasm. TAG Theo. Appl.Genet., 98: 1125-1131.
Hasnaoui N, Buonamici A, Sebastiani F, Mars M, Trifi M, Vendramin GG. 2010b. Development and characterization of SSR markers for pomegranate (Punica granatum L.) using an enriched library. Conservation. Genet. Resour., 2: 283–285
Hasnaoui N, Mars M, Chibani J, Trifi M. 2010a. Molecular Polymorphisms in Tunisian Pomegranate (Punica granatum L.) as Revealed by RAPD Fingerprints. Diversity. 2: 107–114 Jbir R, Hasnaoui N, Mars M, Marrakchi M, Trifi M. 2008.
Characterization of Tunisian pomegranate (Punica granatum L.) cultivars using amplified fragment length polymorphism analysis. Sci. Hort., 115: 231–237
Koohi-Dehkordi M, Seyed-Tabatabaei BE, Yamchi A, DaneshShahraki A. 2007. Microsatellite markers in pomegranate. Acta. Hort., 760: 179–183
Mars M, Marrakchi M. 2000. Diversity of pomegranate (Punica granatum L.) germplasm in Tunisia. Gene. Resour. Cro. Evolu., 46: 461–467
Marshall TC, Slate J, Kruuk L, Pemberton JM. 1998. Statistical confidence for likelihood-based paternity inference in natural populations. Mol. Ecol., 7: 639–655
Moslemi M, Zahravi M, Bakhshi Khaniki G. 2010.Genetic diversity and population genetic structure of pomegranate (Punica granatum L.) in Iran using AFLP markers. Sci. Hort., 126: 441– 447
Nafees M, Jaskani JM, Ahmed S, Awan FS. 2015. Morphomolecular characterization and phylogenetic relationship in pomegranate germplasm of Pakistan. Pak. J. Agri. Sci., 52: 97– 106
Narzary D, Rana TS, Ranade SA. 2010. Genetic diversity in intersimple sequence repeat profiles across natural populations of Indian pomegranate (Punica granatum L.). Plant. Biol., 12: 806–813
Noormohammadi Z, Parvini F, Sheidai M, Vazifeshenas MR. 2010. Further study of morphological and molecular diversity in 18 pomegranate landraces of Iran. Gene Conserve., 9: 189–200
Noormohammadi Z, Fasihee A, Homaee-Rashidpoor S, Sheidai M, Baraki SG, Mazooji A, Tabatabaee-Ardakani SZ. 2012. Genetic variation among Iranian pomegranates (Punica granatumL.) using RAPD, ISSR and SSR markers. Australian. J Crop. Sci., 2: 268–275
Owais SJ, Abdel-Ghani AH. 2016. Evaluation of Genetic Diversity among Jordanian Pomegranate Landraces by Fruit Characteristics and Molecular Markers 2. Int. J. Agric. Biol., 18: 393-402.
Parvaresh M, Talebi M, Sayed-Tabatabaei B. 2012. Molecular diversity and genetic relationship of pomegranate (Punica granatum L.) genotypes using microsatellite markers. Sci. Hortic., 138: 244–252
Pirseyedi SM, Valizadehghan S, Mardi M, Ghaffari MH, Mahmoodi P, Zahravi M, Zeinalabedini M, Khayam Nekoui SM. 2010. Isolation and Characterization of Novel Microsatellite Markers in Pomegranate (Punica granatum L.). Int. J. Mol. Sci., 11: 2010–2016
Prevost A, Wilkinson MJ. 1999. A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor. Appl. Genet., 98: 107–112
Rohlf FG. 2000. NTsys-pc numerical taxonomy and multivariate system version 2.0 Applied. Biostatistics. Inc, New York, USA
Sarkhosh A, Zamani Z, Fatahi R, Hassani ME, Wiedow C, Buck E, Gardiner SE. 2011. Genetic diversity of Iranian soft-seed pomegranate genotypes as revealed by fluorescent-AFLP markers. Physi. Mol. Bio. Plant., 17: 305–311
Sheidai M, Noormohammadi Z. 2005. Chromosome pairing and unreduced Gamete formation in nineteen pomegranate (Punica granatum L.) cultivars. Cytologia., 70: 257–265
Sheidai M, Noormohammadi Z, Saneghi A, Shahryari ZH. 2007. RAPD analysis of eleven Iranian pomegranate (Punica granatum L.) cultivars. Acta. Biol. Szeged., 51: 61–64
Shukla M, Gupta K, Rasheed Z, Khan KA, Haqqi TM. 2008. Bioavailable constituents or metabolites of pomegranate (Punica granatum L.) preferentially inhibit COX2 activity ex vivo and IL-1beta-induced PGE2 production in human chondrocytes in vitro. J. Inflammation., 5: 1–9
Sneath PHA, Sokal RR.1973. Numerical Taxonomy. WH Freeman, San Francisco Soriano JM, Zuriaga E, Rubio P, Lla´cer G, Infante R, Badenes ML. 2011. Development and characterization of microsatellite markers in pomegranate (Punica granatum L.). Mol. Bre., 27: 119–128
Talebi Badaf M, Baharmasoud SB, Yamchi A. 2011. Evaluation of genetic diversity among Iranian pomegranate (Punica granatum L.) cultivars using ISSR and RAPD markers. J. Taxo. Bios., 8: 33–44
Vroh Bi L, Harvengt A, Chandelier G, Mergeai P, Jardin Du. 1996. Improved RAPD amplification of recalcitrant plant DNA by the use of activated charcoal during DNA extraction. J Plant. Bre., 155: 205–206
Yeh FC, Yang RC, Boyle T. 1997. Population Genetic analysis (POPGENE 1.31). A joint project of University of Alberta and Center for International Forestry Research.
Yuan Zh, Yin Y, Qu J, Zhu L, Li Y. 2007. Population genetic diversity in Chinese pomegranate (Punica granatum L.) cultivars revealed by fluorescent-AFLP markers. J Genet. Genom., 34: 1061–1071
Zamani Z, Adabi M, Khadivi-Khub A. 2013. Comparative analysis of genetic structure and variability in wild and cultivated pomegranates as revealed by morphological variables and molecular markers. Plant. Syst. Evol., 299: 1967–1980.