MORPHOLOGICAL AND GENETIC CHARACTERIZATION OF VARIOUS Apis SPECIES CAPTURED FROM SELECTED SITES OF PUNJAB.

The specimens of Apis mellifera, A. dorsata and A. florea were collected during field surveys during July, 2018 to August, 2020 from selected sites of Punjab, Pakistan. These sites included Bhawalpur, Khaniwal, Kasur, Lahore, Chakwal and Rawalpindi. The captured specimens were identified first on the basis of morphology and then identified using mitochondrial DNA genes. A. mellifera workers, queens and drones were black in color. Apis dorsata workers were yellow, however, queens and drones were brown. Apis florea queens and workers were yellow banded and the drones were black. The phenol chloroform technique was used to extract whole genomic DNA from preserved honey bee tissues. The quality of the DNA was determined using gel electrophoresis, and the quantity was determined using a thermal scientific Nano-drop. COI primers were used for PCR amplification. All of the acquired specimens' DNA sequences revealed reliable and obvious species identification. GenBank was contacted when appropriate amplified DNA samples were uploaded, and accession numbers were acquired (MZ433243.1, MZ433277.1 and MZ433289.1). After removing problematic bases, the Apis mellifera, Apis dorsata, and Apis florea COI gene fragments were 670, 680, and 675, respectively. Closely comparable Apis sequences were collected from NCBI in blast queries and used in phylogenetic studies. Neighbor-Joining tree, Maximum Likelihood tree and Minimum Evolution tree of the genus Apis was constructed based on p-distance using MEGA X. The overall, genetic divergence of genus Apis was 0.098 ± 0.022. It can be concluded that the morphological characteristics provide information as an indicator for estimating genetic variations between honey bee populations. Further work is needed to explore more areas of Pakistan and molecular identification is required to report any new or subspecies from country. [ABSTRACT FROM AUTHOR]