Molecular Identification and Phylogenetic Analysis of Distinct Geographical Populations of Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) in Pakistan

The morphological identification using diagnostic morphological keys need tedious procedure of microscopic examination of male genital structures of adults and also require preparation of sample or specimens. Molecular identification and characterization for invasive insect pest based on mitochondrial cytochrome oxidase I (COI) and Inter transcribed spacer (ITS) is very important for correct identification. Red Palm Weevil (RPW) (Rhynchophorus ferrugineus) is an invasive palm pest and its mitochondrial cytochrome oxidase I (COI) and Inter transcribed spacer (ITS) based identification and genetic variation is very important because of its high polymorphism and existence of different Rhynchophorus spp. Here, for the first time, based on both COI and ITS primers, we have identified and characterized Red Palm Weevil (RPW) (R. ferrugineus) collected from four Provinces (KPK, Baluchistan, Sindh and Punjab) of Pakistan. The genetic analysis of various geographical population of Red Palm Weevil (RPW) (R. ferrugineus) was carried out through PCR. Phylogenetic analysis of nucleotides sequence done by framing maximum likelihood trees using MEGA6 indicated that there were no genetic variations among our R. ferrugineus strains showing 99-100% among themselves while in association with other Rhynchophorus strains reported from KSA, UAE, Greece and USA exposed 97 to 99% homology. Further, RPW population collected from 4 provinces of Pakistan shared 84, 85 and 87% homology for R. palmarum, R. vulneratus and R. cruentatus respectively. While, the evolutionary genetic divergence of our R. ferrugineus strains RPW1 (Acc. no: KY458178), RPW3 (Acc. no: KY458179) and RPW4 (Acc. no: KY458180) with other R. ferrugineus strains of other countries ranged 2-10 indicating maximum similarity index. We found value greater than 10 of evolutionary divergence for strains other than R. ferrugineus reported from other countries. This is the first evidence of R. ferrugineus widely occurrence, and its molecular identification and characterization from four provinces of Pakistan. This study will help us to develop species specific R. ferrugineus control measures. (C) 2018 Friends Science Publishers