Genetic relationship and diversity analysis in four buffalo breeds of Pakistan.

Although, the water buffalo (Bubalus bubalis) is a backbone of the Pakistani dairy industry but a very limited knowledge at molecular and genetic level is available. Hence, the objective of current study was to analyze the genetic diversity of Pakistani buffaloes. Seventy D-loop partial sequence (880-bp) haplotypes with 71 polymorphic sites were detected among 212 individuals, collected form Punjab and Sindh provinces of Pakistan, mtDNA d-loop region was first amplified and sequenced. The UPGMA phylogenetic tree revealed that, the Ancient Pakistani buffalo Glade was more old and ancient as compare to Indian Riverine buffalo Glade. Whereas, the haplotype and nucleotide diversity (0.9561 +/- 0.00010 and 0.00988 +/- 0.00060) was higher in the Nili-Ravi, followed by Kundhi (0.9386 +/- 0.00020 and 0.01043 +/- 0.00083), Ravi (0.9130 +/- 0.016 and 0.01012 +/- 0.00112) and Nili (0.8874 +/- 0.054 and 0.00999 +/- 0.00164), respectively. Median Joining Network also disclosed two different clades in only Pakistani buffaloes, Ancient Pakistani buffalo and Sister Pakistani buffalo Glade same as in phylogenetic tree. However, Pakistani and Indian Riverine buffaloes did not share any haplotype with each other. The buffalo populations were separated from each other through of (>50 mutational steps). Further, the Ancient Pakistani Glade showed high time depth in phylogenetic tree by residing close to the base root and highest Ancestry in Median Joining network by having the most numbers of mutational steps as compared to other two Riverine buffalo clades. Our findings indicated that the Pakistani Riverine buffaloes diverged earlier. The genetic information obtained from the mtDNA d-loop is of utmost importance for the future genetic studies and improvements.