Phylogeny and diversity study in Damani sheep through mitochondrial DNA D-loop nucleotides sequence

The current study is the first of its sort to evaluate the mitochondrial DNA (mtDNA) control region i.e. Displacement Loop (D-loop) in Damani sheep breed of Khyber Pakhtunkhwa Province, Pakistan. Limited work had been done on Damani sheep being recognized based on morphology and SSRs (Same Sequence Repeats) markers, which have not provided information about its origin. Thus, the current study was designed to investigate genetic diversity, relationship and matrilineal lineage of Damani sheep. Thirty (30) pure breed Damani sheep male and female were selected for blood sampling. The DNA was extracted from blood samples using Non-enzymatic salting out method and then (PCR) Polymerase Chain Reaction was performed for amplification of D-loop 910 bp region. The amplicons were purified and then sequenced using Sanger sequencing technique. The sequences of D-loop revealed sixty-seven (67) polymorphic positions which were further subdivided into six haplotypes. The mean composition of D-loop nucleotides sequences were A/T and G/C contents (62.08 and 38.56%, respectively). Mean haplotype (HD) and nucleotides diversity (π) of D-loop was 0.853, and 0.02867, respectively. Two main clades were detected in Phylogenetic tree of D-loop sequences including the wild sheep (Ovis ammon) and domestic sheep (Ovis aries). The clustering pattern of Damani sheep D-loop haplotypes showed common matrilineal lineage with Chinese and Russian native sheep further it was concluded that population of this breed harbors considerable genetic diversity. The findings of this study will be a baseline for effective breeding strategies and conservation management of this breed in future.