mtDNA analysis
A total 140 samples (five samples per population) were used forCOI and cyt b analysis. PCR primers CB1 (5’-TATGTACTACCATGAGGACAAATATC-3’) and CB2 (5’-ATTACACCTCCTAATTTATTAGGAAT-3’) were used to sequence 428 bp region of mitochondrial cyt b region (Jermiin & Crozier, 1994) while LCO1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HC02198 (5’-TAAACTTCAGGGTGACCAAAAAATCA-3’) primers were used to sequence 625 bp region of mitochondrial COI region (Folmer, Black, Hoeh, Lutz, & Vrijenhoek, 1994). PCR amplifications were conducted with a 3 min denaturation at 94 °C followed by 35 cycles of
45 s denaturation at 94 °C, 1 min at 48 oC forcyt b and 47 oC for COI , 1.5 min at 72 °C. The final extension was applied at 72 °C for 10 min. Amplified products were purified and sequenced using forward and reverse primers.