Multiplex PCR-based identification of two sympatric stem borer species, <i>Sesamia cretica</i> and <i>Sesamia nonagrioides</i> (Lepidoptera: Noctuidae)

Stem borers of the genus Sesamia are major pests of cereals in tropical regions. In Iran, Sesamia cretica and Sesamia nonagrioides significantly affect maize and sugarcane, but distinguishing them at the larval stage is difficult due to morphological similarity. In this study, we developed and optimized a rapid, cost-effective multiplex PCR method for simultaneous identification of both species. Primers designed from GenBank sequences yielded specific bands of 367 bp for S. cretica and 438 bp for S. nonagrioides. PCR and sequencing confirmed the specificity of the designed primers, while morphological comparison of larvae and reared adults validated the molecular results. This molecular tool supports integrated pest management by improving identification accuracy, which could help reduce unnecessary pesticide applications.