DNA barcoding of Tribolium castaneum (Coleoptera: Tenebrionidae) from selected states in Nigeria based on mitochondrial DNA sequences

Abstract

Tribolium castaneum also known as red flour beetle is one of the most important pests of stored grain product with a cosmopolitan distribution in Nigeria and all over the world contributing to food spoilage. The aim of this study was to characterize the T. castaneum by morphometric and molecular analyses. Samples of yam flour with evidence of the red flour beetles present inside were obtained from four locations in Kwara, Kogi, Oyo and Ekiti states in Nigeria. Morphological and molecular identifications of T. castaneum were carried out using standard methods. A dissecting microscope was used to identify the beetles and measurements were taken using ImageJ. Genomic DNA was extracted and checked on 1.5% agarose gel to confirm the presence of DNA. Species-specific primers were used to amplify mitochondrial cytochrome oxidase I (COI) gene of T. castaneum and the PCR amplicon size was also checked on 1.5% agarose. Morphometric measurements showed that the highest mean number (33.00±4.24 mm) of T. castaneum larvae observed was recorded on day 61 in Ilorin and the lowest was in Iwo, Osun state (4.00±0.00 mm) on the same day. The mean of the total body length of larvae from sampling sites was (1.31±0.37 mm) with minimum and (1.63±1.14 mm) maximum lengths respectively. There was no significant difference (p>0.05) between the mean length of the larvae collected from the study locations. Aligned cytochrome oxidase subunit 1 (COI) sequences of 313bp were analyzed. Phylogenetic analysis inferred by maximum likelihood method showed that the T. castaneum sequences analyzed for this study and sequences obtained from GenBank formed a monophyletic group. The molecular and phylogenetic analyses confirmed the presence of a single species of T. castaneum. The results from this study showed low levels of genetic diversity and variability in the studied T. castaneum populations. The observed genetic similarity in T. castaneum could be due to the fact that they were probably from similar origin when compared with those in the GenBank database. However, further studies are needed with more samples to characterize T. castaneum species from stored food grains across Nigeria.

Keywords

• COI gene
• mitochondrial DNA
• PCR
• Tribolium