Traditional techniques for identifying the red cotton bugs Dysdercus koenigii and Dysdercus cingulatus (Pyrrhocoridae: Hemiptera) depend on phenotypic and morphological data. These methods are suitable when species have well-defined structures and when huge taxonomic knowledge is at hand. In this study, the mitochondrial cytochrome oxidase I (COI) gene was selected for the molecular identification of D. koenigii and D. cingulatus. The sequence data showed that one species of the red cotton bug

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... ample was identified as Dysdercus koenigii with 35-100% similarity to other Dysdercus species present in the NCBI database, while the other species was identified as Dysdercus cingulatus with 88-94% similarity to other red cotton bugs species present in the NCBI database. The phylogenetic analysis revealed that our D. koenigii Seq (>180319003-A02-102-DK-.ab1) shares the same cluster with four D. koenigii isolates (ZSI/SRC_I.28B, ZSI/SRC_I.28, GQ306227.1, KJ459924.1). While the D. cingulatus Seq (>180319003-A02-103-DC-.abl) shares a cluster with D. cingulatus voucher (RO_DC2015). This is the first molecular identification of red cotton bugs (D. koenigii and D. cingulatus) reported in Pakistan. Thus, in the current study, molecular identification has been accepted as a reliable method for the identification of these agriculturally important insect pests.