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DNA BARCoding to Resolve the Confusion in Identifying Tribolium Confusum and Tribolium Castaneum

Abu Faiz Md Aslam
Department of Zoology, Jahangirnagar University, Savar, Dhaka -1342, Bangladesh.

Sharmin Sultana
Department of Zoology, Jahangirnagar University, Savar, Dhaka -1342, Bangladesh.

Sumita Rani Das
Department of Zoology, Jahangirnagar University, Savar, Dhaka -1342, Bangladesh.

Abdul Jabber Howlader
Department of Zoology, Jahangirnagar University, Savar, Dhaka -1342, Bangladesh.

Abstract/ Executive Summary:

Tribolium confusum and Tribolium castaneum (Coleoptera: Tenebrionidae) are two very confusing pest species while identification is done on the basis of morphology only. Such pests are discovered in stored grain as immature stages, which further complicates the identification process. Accurate identification of these pests is urgently required for integrated pest management. In this research, DNA barcoding was used to identify these pests accurately at any life stage. A 658 bp fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene was analyzed. DNA barcode datasets of T. confusum (GeneBank Acc. no. MK120453.1) and T. castaneum (Acc. no. MK411585.1) were constructed. The nucleotide composition reveals that average AT contents (59.9%) were higher than the GC contents (38.6%). Phylogenetic analysis by the maximum likelihood method showed that both the species were originated from a common major clade. About 17.13% nucleotide differences were noted between the CO1 sequences by multiple sequence alignment. The interspecies nucleotide genetic distance (0.200) was calculated using Kimura 2 parameter. Haplotype analysis showed high genetic diversity (112 mutational steps) among them.

Keywords: Tribolium confusum, T. castaneum, COI gene, DNA barcoding

Location:

Start Date:

End Date:

Program Area: Pest Management

Commodity: DNA, Pest

Objectives:

To identify Tribolium confusum and Tribolium castaneum (Coleoptera Tenebrionidae): these pests are very confusing species that are required for integrated pest management. DNA barcoding was used to identify these pests accurately at any life stage

Methodology:

Insect collection: The populations of Tribolium confusum and T. castaneum used in this study were collected from the laboratory colony maintained in the Molecular Entomology Laboratory, Department of Zoology, Jahangirnagar University, Savar, Dhaka, Bangladesh. Insects were reared on flours at 25⁰C and 65 - 70% RH. Genomic DNA extraction: The genomic DNA was extracted from somatic tissue rich in mitochondria (e.g., leg or wing) using Wizard® Genomic DNA Purification Kit, USA, following the manufacturer’s protocol as briefly mentioned in Aslam et al. (2019). Tribolium flour beetles (Coleoptera : Tenebrionidae) are cosmopolitan pests of stored grains, and they are among the most important insect pests in grain processing and other storage facilities (Sokoloff 1972, Campbell et al. 2004). Flour beetles decrease the quality and quantity of stored products by introducing insect fragments and excrement (Nowaczyk et al. 2009) as well as causing contamination with potential adverse effects to human health (Stejskal and Hubert 2008). The genus Tribolium has 36 described species (Angelini and Jockusch 2008), 10 of which are stored products pests (Nakakita 1983). In Bangladesh, two species of flour beetles (T. casteneum and T. confusum) are commonly found (Banglapedia 2007). The red flour beetle, Tribolium castaneum (Herbst), and the confused flour beetle, T. confusum (Jacquelin du Val) (Coleoptera : Tenebrionidae), are sympatric and morphologically similar stored-product pests, and the latter is named because of confusion over its identity as it is so similar to the former (Walter 1990). These two pests are differently susceptible to different insecticides (Arthur 2008). Also, immature stages (larvae and pupae) of these pests are more susceptible compared to the adult stages (Arthur and Campbell 2008). To reduce the use of insecticides and improve the level of pest management, it is important for a pest manager to identify them properly. In routine stored-grain insect pest monitoring and port quarantine, it is difficult to identify species. These pests are usually found as body parts or immature stages, making it virtually impossible to identify the species on the basis of external morphological characteristics alone. However, T. confusum and T. castaneum are indistinguishable in the egg, larval and pupal stages, and difficult to distinguish in the adult stages without professional training. Therefore, it is necessary to develop a useful alternative method to identify the two sibling flour beetle species (Ming et al. 2014).

Source of Information: Bangladesh J. Zool. 47(2): 333-342, 2019

Article Link (Online Journal): DOI: https://doi.org/10.3329/bjz.v47i2.44344

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Achievement/Findings:

Overall, these findings will contribute to a better understanding of the identification of T. confusum and T. castaneum pests by COI gene. Moreover, DNA barcoding can help in identifying pests at any stage of their life, making it easier to control them and saving farmers from the cost of billion dollars from pest damage.

Knowledge Management: Journal