Polymerase chain reaction (PCR) based molecular characterization of popular wheat varieties of Khyber Pukhtunkhwa (KPK) region of Pakistan

Abstract

Wheat breeders in Pakistan relied upon phenotypic traits as indices of genetic variation among different wheat varieties which may genotypically be closely related or vice versa. The main objective of the current study was to determine the genetic diversity among different popular wheat cultivars grown in the Khyber Pukhtunkhwa (KPK) region of Pakistan, which was previously not fully understood. 13 wheat cultivars namely Zarghoon, Haider-2000, Ghaznavi-98, Nowshera-96, Khyber-79, Dera-98, Tatara, Khatakwal, Saleem-2000, Fakhre Sarhad, Pirsabak-85, Takbeer and Inqilab-91 were used in the current study. Deoxyribonucleic acid (DNA) was isolated using standard protocols and procedures. Polymerase chain reaction (PCR) was carried out using randomly amplified polymorphic DNA (RAPD) primers to amplify DNA of all wheat genotypes. PCR product were electrophoresed on 2% agarose gel and visualized by staining with ethidium bromide under UV light. Unweighted pair group of arithmetic means (UPGMA) function was used to estimate genetic distance using bivariate data set generated from all loci of RAPD primers. Cluster analysis carried out and dendrogram was generated using computer program PopGene-32 on similarity matrix. The RAPD markers used in this study identified high rate of genetic polymorphism. Of these primers GLA-08 showed highest genetic diversity (43.42%), while GLA- 02 showed lowest genetic diversity (31.73%). Four major groups were identified amongst wheat genotypes. Fakhre Sarhad-Tatara and Khatakwal-Takbeer were found to be genetically closely related with a genetic distance of 6.4 and 9.7%, respectively. Zarghoon and Khatakwal are genetically very distant with a genetic distance of 64.8%. Molecular markers used in this study show high rate of genetic diversity that can be used to assist a breeding program for the improvement of wheat in KPK-Pakistan.

Key words: Wheat, molecular characterization, KPK, genetic distance.