Evaluation of Phylogenetic Characteristics of Iranian Honeybee (Apis mellifera meda) Populations based on Mitochondrial ND2 Gene

For the identification of phylogenetic characteristics of honeybee populations, sampling was conducted from 31 provinces of Iran in spring and summer 2016. Phylogenetic characteristics were evaluated based on mitochondrial ND2 gene. The intergenic regions between ND2 and COI genes were compared in different populations of honeybees. After sequencing and alignment of the genes, the relationships among populations were analyzed by MrBayes 3.2 and PAUP 4.0 b10 softwares. Eight nucleotide differences were found among Iranian populations of honey bee (A. m.meda). The phylogenetic tree was drawn and Iranian populations of honeybee (A.m.meda) were divided into four groups. The results showed that samples of East Azarbaijan and Yazd were separated from other honeybee populations. In addition, these two honeybee populations had the highest intergenic region (ITS2 with 70 nucleotides). Not only, populations of Charmahal Bakhtiari, Tehran, Sistan and Blochestan, Mazandaran, Lorestan, Kordestan, Kermanshah, Kohkeloye and Boyerahmad, Southern Khorasan, Ilam, Golestan and Gazvin were grouped with each other but also, ITs2 lengths of these populations were 62 bp. ITs2 Lengths were 59 to 70 bp. ITs1 lengths were 20 bp except syriaca subspecies. The shortest length of intergenic region was related with ITs3 (with two nucleotides AT). Populations of Ardabil, Zanjan and Kerman were grouped with bootstrap of 91 percent. Additionally, populations of Alborze, Northern Khorasan, Razavi Khorasan, Esfahan, Shiraz, Semnan, Markazi, Khozestan, Hormozgan, Hamedan, Qom, Boshehr and Western Azarbayejan were grouped with each other. The honeybee populations were compared using two-parameter Kimura method. Results demonstrated that there was no nucleotide difference between Gilan population and A. m.carnica subspecies. The collected samples from Gilan were not A.m.meda subspecies and were grouped with A.m.carnica subspecies. A.m.carnica is not a native subspecies therefore honeybee queens have been imported illegally by some beekeepers. A.m.intermissa and A.m.scutellata showed the most genetic distance (0.01) in comparison with Iranian populations of honeybee (A.m.meda). Population comparisons of Alborz, Shiraz, Semnan, Markazi, Khozestan, Hormozgan, Hamedan, Qom, Boshehr and Western Azarbayejan showed that there was no genetic difference among populations. The phylogenetic tree could differentiate syriaca, intermissa, scutellata and mellifera subspecies from carnica and meda subspecies based on ND2 gene. Moreover, A.m.ligustica was differentiated from A.m.carnica with a substitution C→T.