Identification of Genetic Variation of Male and Female Date Palm (Phoenix dactylifera L.) Cultivars Using Morphological and Molecular Markers

Document Type: Research paper


1 Horticulture Crops Research Department, Hormozgan Agricultural and Natural Resources Research and Education Center, AREEO, Bandar Abbas, Iran

2 Department of Plant Protection, Hormozgan Agricultural and Natural Resources Research Center, Bandar Abbas, Iran


Date palm (Phoenix dactylifera L.) is a monocot, dioecious and high longevity tropical plant greatly considered due to its high economic importance in Middle East. This investigation was carried out to detect genetic relations among 34 cultivars of Iranian date palm, both males and females, using ISSR markers. Accordingly, total DNA was extracted using CTAB method and was subjected to PCR amplification using 12 ISSR primers. Analysis of ISSR data indicated that PIC value varied from 0.3695 to 0.4998 with average of 0.4497. The highest PIC value belonged to (CT)10 G primer (0.4498). Totally, 112 alleles were recognized. According to morphological attributes and ISSR markers, the 34 studied date cultivars divided into eight main groups and different sub-groups, and no significant correlation between grouping made by morphological and molecular markers. Unlike ISSR markers that produced a clear-cut among male and female cultivars, no distinct border was seen among male and female cultivars using morphological attributes. Furthermore, PCA analysis confirmed the grouping made by ISSR markers. In conclusion, ISSR markers have been useful to separate date palm cultivars.


Adawy S.S, Hussein EHA, El-Khishin D, Saker M.M, El-Itriby H.A. 2002. Genetic variability studies and molecular fingerprinting of some Egyptian date palm (Phoenix dactylifera L.) cultivars II-RAPD and ISSR profiling. Arab Journal of Biotechnology 5, 225-236.
2. Adawy S.S, Hussein E.H.A, El-Khishin D, Saker M.M, Mohamed A.A, El-Itriby H.A. 2011. Genotyping Egyptian date palm cultivars using RAPD, ISSR, AFLP markers and estimation of genetic stability among tissue culture derived plants. From: Datepalm/Topics/ Biotech/Biotech-right.htm.
3. Ahmed T.A, Al-Qaradawi A.Y. 2010. Genetic diversity of date palm genotypes in Qatar as determined by SSR and ISSR markers. In Proceeding of 4th International Date Palm Conference, Eds.: Zaid A and Alhadrami GA, Acta Horticulture 882, ISHS.
4. Aitchitt M, Ainsworth C.C, Thangavelu M. 1993. A rapid and efficient method for the extraction of total DNA from mature leaves of the Date Palm (Phoenix dactylifera L.). Plant Molecular and Biology Reports 11(4), 317-319.
5. Al-Issa A.M, Al-Helal A.A, Al-Saad F.A. 2008. DNA fingerprinting analysis for three date palm Phoenix dactylifera L. cultivars grown at Al-Ahsa and Al-Qatif in Saudi Arabia using (ISSR-PCR) technique. Magazine of Abd-Al-Aziz King, Astro and Environmental Science and Agriculture in Dry Lands 19, 3-21.
6. Al-Khalifa N.S, Askari E. 2003. Molecular phylogeny of date palm (Phoenix dactylifera L.) cultivars from Saudi Arabia by DNA fingerprinting. Theoretical and Applied Genetics 107, 1266-1270.
7. Al-Mahmoud M.E, Al-Dous E.K, Al-Azwani E.K, Malek J.A. 2012. DNA-based assays to distinguish date palm (Arecaceae) gender. American Journal of Botany e7-e10.
8. Anonymous. 2016. Agriculture statistics. Vol. III: Horticultural Crops. Jahad-Keshavarzi Ministry. (In Persian)
9. Arabnezhad H, Bahar M, Mohammadia H.R,
Identification of Genetic Variation of Male and Female Date Palm … 75
Latifianb M. 2012. Development, characterization and use of microsatellite markers for germplasms analysis in date palm (Phoenix dactylifera L.). Scientia Horticulturae 134, 150-156.
10. Arslan E, Tamkoc A. 2011. The application of ISSR-PCR to determine the genetic relationship and genetic diversity between narrow leaved Bluegrass (Poa angustifolia) and rough bluegrass (Poa trivialis) accessions. Turkish Journal of Biology 35, 415-423.
11. Bruschi P, Vendramin G.G, Bussotti F. 2003. Morphological and molecular diversity among Italian populations of Quercus petraea (Fagaceae). Annual Botany 91, 707-716.
12. Corniquel B, Mercier L. 1994. Date palm (Phoenix dactylifera L.) cultivar identification by RFLP and RAPD. Plant Science 101, 163-172.
13. Cullis C. 2011. Molecular markers in date palm. In: S.M. Jain et al. (eds.), Date palm biotechnology 361-370.
14. Dhawan C, Kharb P, Sharma R, Uppal S, Aggarwal RK. 2013. Development of male-specific SCAR marker in date palm (Phoenix dactylifera L.). Tree Genetics and Genomes DOI: 10.1007/s11295-013-0617-9.
15. Eissa E.A, Abdel-razek B, El-sharabasy F, Rizk R.M. 2009. Morphological and molecular genetic characterization of soft date palm (Phoenix dactylifera L.) cultivars in Egypt. Egyptian Journal of Genetics and Cytology 38, 269-284.
16. Gürcan K, Mehlenbacher S.A, Cristofori V. 2009. Inter-simple sequence repeat (ISSR) markers in hazelnut. Acta Horticulturae (ISHS) 845, 159-162.
17. Haider N, Nabulsi I, MirAli N. 2012. Phylogenetic relationships among date palm (Phoenix dactylifera L.) cultivars in Syria using RAPD and ISSR markers. Journal of Plant Biology Research 1(2), 12-24.
18. Hamza H, Benabderrahim M.A, Elbekkay M, Ferdaous G, Triki T, Ferchichi A. 2012. Investigation of genetic variation in Tunisian date palm (Phoenix dactylifera L.) cultivars using ISSR marker systems and their relation with fruit characteristics. Turkish Journal f Biology 36, 449-458.
19. Hashempour, M. (1999). Date depository. Vol. I. Agriculture Education Press. (In Persian)
20. Hassanzadeh Khankahdani H, Rastegar S, Golein B, Golmohammadi M, Aboutalebi Jahromi A. 2018. Genetic variation of lime (Citrus sp.) accessions using flow cytometry technique,
morphological characteristics and molecular markers. International Journal of Horticultural Science and Technology 5(2): 199-208.
21. Hussein E.H.A, Adawy S.S, Ismail S.E, El-Itriby H.A. 2005. Molecular characterization of some Egyptian date palm germplasms using RAPD and ISSR markers. Arab Journal of Biotechnology 8(1), 83-98.
22. Karim K, Chokri B, Amel S, Wafa H, Richid H, Nouredine D. 2010. Genetic diversity of Tunisian date palm germplasms using ISSR Markers. International Journal of Botany 6(2), 182-186.
23. Khajepour-Tadovani A, Arzani K, Zargari H, Sarikhani-Khorami S. 2016. Effect of pollen grain on quantitative and qualitative characteristics of date fruit of Shahani cultivar. Seed and Plant Breeding Journal 32-1(3), 293-310. (In Persian)
24. Khanam S, Sham A, Bennetzen J.L, Aly M.A.M. 2012. Analysis of molecular marker-based characterization and genetic variation in date palm (Phoenix dactylifera L.). Australian Journal of Crop Science 6(8), 1236-1244.
25. Maraashi S.S. 2006. National prescription of distinction, uniformity and stability (DUS) tests in date palm. Seed and Plant Certification and Registration Institute 41 p. (In Persian)
26. Marsafari M, Ashraf-Mehrabi A. 2013. Molecular identification and genetic diversity of Iranian date palm (Phoenix dactylifera L.) cultivars using ISSR and RAPD markers. Australian Journal of Crop Science 7(8), 1160-1166.
27. Mirbahar A.A, Markhand G.S, Khan S, Abul-soad A.A. 2014. Molecular characterization of some Pakistani date palm (Phoenix dactylifera l.) cultivars by RAPD markers. Pakistan Journal of Botany 46(2), 619-625.
28. Mitra C, Kharb P, Uppal S. 2011. Genetic diversity analysis in date palm (Phoenix dactylifera L.): a comparative assessment using ISSR and RAPD marker assays. Journal of Horticultural Science and Biotechnology 86, 398-402.
29. Moghaieb R.E.A, Abdel-Hadi A.A, Ahmed M.R.A, Hassan A.G.M. 2010. Genetic diversity and sex determination in date palms (Phoenix dactylifera L.) based on DNA markers. Arab Journal of Biotechnology 13(2), 143-156.
30. Munshi A, Osman G. 2010. Investigation on molecular phylogeny of some date palm (Phoenix dactylifra L.) cultivars by protein, RAPD and ISSR markers in Saudi Arabia. Australian Journal of Crop Science 4(1), 23-28.
76 Int. J. Hort. Sci. Technol; Vol. 6, No. 1; June 2019
31. Murray H.G, Thompson W.F. 1980. Rapid isolation of high molecular weight DNA. Nucleic Acids Research 8, 4321-4325.
32. Naghavi M.R, Ghareyazi B, Hosseini-Salekdeh B. 2005. Genetic principles. Tehran University Publication. (In Persian)
33. Pasqualone A, Lotti C, Bruno A. 2000. Use of ISSR markers for cultivar identification in durum wheat. Option Mediterranean Series A 40, 157-161.
34. Rezazadeh R, Hassanzadeh H, Hosseini Y, Karami Y, Williams R.R. 2013. Influence of pollen source on fruit production of date palm (Phoenix dactylifera L.) cv. Barhee in humid coastal regions of southern Iran. Scientia Horticulturae 160, 182-188.
35. Rhouma S, Zehdi SA, Ould Mohamed Salem A. 2007. Genetic diversity in ecotypes of Tunisian date palm (Phoenix dactylifera L.) assessed by AFLP markers. Journal of Horticultural Science and Biotechnology 82, 929- 933.
36. Rhouma S. 2008. Analyse de la diversite genetique chez le palmier dattier (Phoenix dactylifera L.). Etude transcriptomique de la maladie des feuilles cassantes. Ph.D, University of Tunis-El Manar.
37. Rhouma-Chatti S, Baraket G, Dakhlaoui-Dkhil S. 2011. Molecular research on the genetic diversity of Tunisian date palm (Phoenix dactylifera L.) using the random amplified microsatellite polymorphism (RAMPO) and amplified fragment length polymorphism (AFLP) methods. African Journal of Biotechnology 10, 10352-10365.
38. Sakka H, Zehdi S, Ould Mohamed Salem A. 2003. Tunisian date-palm (Phoenix dactylifera L.) genotypes identification mediated by plastid PCR/RFLP based DNA. Journal of Genetic and Breeding 57, 259-264.
39. Sedra M.H, Lashermes H.P, Trouslot P. 1998. Identification and genetic diversity analysis of date palm (Phoenix dactylifera L.) varieties from Morocco using RAPD markers. Euphytica 103, 75-82.
40. Snoussi H, Du Jardin P, Ben Abdallah A. 2001. Assessment of genetic variation within date palm (Phoenix dactylifera L.) using amplified fragment length polymorphism (AFLP)- genotyping of apomictic seedlings as a case study. In proceeding of 2nd International Conference on Date Palm, Al-Ain United Arab Emirates March 25-27, 678-683.
41. Yusuf A.O, Culham A, Aljuhani W, Ataga C.D, Hamza A.M, Odewale J.O, Enaberue L.O. 2015. Genetic diversity of Nigerian date palm (Phoenix dactylifera) germplasm based on microsatellite markers. International Journal of Bio-Science and Bio-Technology 7(1), 121-132.
42. Zehdi S, Sakka H, Rhouma A, Ould Mohamed S.A, Marrakchi M, Trifi M. 2004a. Analysis of Tunisian date palm germplasms using simple sequence repeat primers. African Journal of Biotechnology 3(4), 215-219.
43. Zehdi S, Trifi M, Billotte N. 2004b. Genetic diversity of Tunisian date palms (Phoenix dactylifera L.) revealed by nuclear microsatellite polymorphism. Hereditas 141, 278-287.
44. Zehdi S, Trifi M, Ould Mohamed Salem A. 2002. Survey of inter-simple sequence repeat polymorphisms in Tunisian date palms (Phoenix dactylifera L.). Journal of Genetics and Breeding 56, 77-83.
45. Zehdi-Azouzi S, Rhouma S, Ould Mohamed Salem A. 2009. Comparative analysis of genetic diversity in Tunisian collections of date palm cultivars based on random amplified polymorphic DNA and inter simple sequence repeats fingerprints. Acta Horticulturae 814, 149-156.