Genetic Diversity and Relationships among Verbascum Species in Iraq by RAPD-PCR Technique

Abstract

Piyada Theerakulpisut, AL-Jewari Hazim, Al-Hadeethy Muazaz, BarusruxSahapat BarusruxSahapat, AL-Mshhdani Athiya, Jawad Mohammed M, AL-Khesraji Talib and Nantawan Kanawapee

This study was aimed to determine the genetic diversity and relationships among 20 local Verbascum species by using the Random Amplified Polymorphic DNA (RAPD) technique. To achieve this, Total genomic DNA of Verbascum plant was extracted from leaves using two different methods, by CTAB method using 2-3 g of leaves, with purity of 1.5-1.9, and also by using commercial kit, PCR reactions were carried out using 17 random decamere-nucleotide primers. Percentage of monomorphic and polymorphic bands varies for each primer used with each species and the lowest numbers of polymorphic bands were 2 with OPX16, which showed the polymorphism (50%), produced 4 bands which only 2 were polymorphic. OPA5 and OPW13 showed the highest number of bands (9) and all are polymorphic (100%). From total of 119 alleles that detected, 103 were polymorphic (86.55%) and 16 monomorphic Moreover. A total of 119 polymorphic bands were scored and used for the analysis of genetic distances. Band sizes varied between 100 and 1900 bp. According to the results of genetic distance and relationships illustrated, the ability to resolve genetic variation among different Verbascum species may relate to the number of polymorphic bands detected with marker technique. Dendrogram was constructed using UPGMA cluster analysis and depicted genetic relationships among 20Verbascum species, showed the 2 major clusters depending on their ancestor and their morphological traits. The information generated from this study can be used in the future for Verbascum breeding and improvement programs.