Genetic Diversity of Nili-Ravi from Nili and Ravi Buffalo Breeds of Pakistan

A genomic study was conducted on three buffalo breeds (Nili, Ravi and Nili-Ravi) by RAPD analyses using 25 random 10-mer primers to identify the genetic diversity among these breeds. Blood samples from forty seven true representative animals of three breeds were collected from their respective home tracts. Six genomic DNA samples from each breed were randomly selected and pooled for analysis. One hundred and forty six bands were observed from 20 primers with an average of 7.3 bands (2-12). Of these 146 DNA bands, 39 fragments (26.71%) showed polymorphism among three buffalo breeds, while the rest 107 bands (73.29%) were monomorphic. The level of DNA variation was observed low. The coefficient of genetic distance among these buffalo breeds of the Punjab province of Pakistan was very low (0.0748 to 0.1312). The maximum genetic distance was found between Nili and Nili-Ravi (0.1312), while, minimum genetic distance was found between Ravi and Nili-Ravi (0.0748). The genetic distance between Nili and Ravi was 0.1298. The survey during blood sampling showed that Nili and Ravi are two separate breeds still found in huge number in their respective home tracts, while the junctures have the new breed named Nili-Ravi. The results suggest that these three breeds must be maintained and improved with their own identity in their native areas

Evaluation of AMPK Genes as Candidates for Production Traits in Buffalo Breeds of Pakistan

The 5’-activated protein kinase (AMPK) also called PRKA is a heterotrimeric protein complex that regulates cellular metabolism in response to nutritional stress in mammals. Total length of AMPK gene is 6500 base pairs. AMPK γ3 has 884 base pairs comprises of exon 3, intron 3 and exon 4. AMPK γ3 gene was studied on DNA from three buffalo breeds (Nili, Ravi and Nili Ravi) of Pakistan. Amplification of this region was done from position 918 in exon to position 1802 just downstream of exon 4, using primers U1A-GAGCAAGGAGACAGCACTTCA (918-938) and U2B-ACCTGTAGCATGGTGTC GAAGA (1781- 1802) with Amplictaq Gold, at annealing temperature 60oC. Sequencing of PRKA γ3 UP SEQ1 5’-GACCTCAGCATCCAGGCT and PRKA γ3 UP SEQ2 5’-GACAGTAACTCCATCTTCCA was done. The results showed that at position 1069, Allelic frequency of C/T was 0.35, T/T was 0.05 and frequency of C/C was 0.40. At position 1343, frequency of A/G was 0.1 and A/A was 0.9. At 1381 position, frequency of C/T was 0.35, T/T was 0.05 and C/C was 0.6. At position 1488, the A/G was 0.35, A/A was 0.05and G/G was 0.6. The detected SNP at 1343 is A/G which is the ancient haplotypes of PRKAG3 in Nili Ravi buffalo breeds from Pakistan. These SNPs in AMPK gene can be used as marker for efficient utilization of energy in these three buffalo breeds of Pakistan that will increase the economic returns to farmers