Molecular identification of Heliothine species by nuclear and mitochondrial PCR-RFLP profile

Abstract

The taxonomy of the Heliothinae subfamily is complex, often becoming an obstacle in the identification of these species, especially during the larval stage. An alternative is the use of molecular methods such as the PCR-RFLP technique (or Restriction Fragment Length Polymorphism), which helps to aid and complement the identification of some heliothine species. The aim of this study was to establish the PCR-RFLP profile using molecular markers for the identification of four heliothine species. Specific primers were constructed for the amplification of the cytochrome oxidase I (COI) and elongation factor 1 alpha (EF-1) genes for identifying Helicoverpa armigera, H. zea, H. gelotopoeon, and Chloridea virescens species. In the digestion of the COI gene with the BfaI enzyme, fragments of 294 bp and 321 bp were obtained for H. armigera, while there was no digestion for H. zea, H. gelotopoeon, and C. virescens. The HpaI enzyme produced fragments of 58 bp and 557 bp for H. gelotopoeon, while there was no digestion for H. armigera, H. zea, and C. virescens. The digestion of the EF-1 gene with the EcoRV enzyme produced fragments of 142 bp and 964 bp for C. virescens, while there was no cleavage for the other heliothines. These molecular markers can help the entomologists, aiding in the discrimination between H. zea, H. armigera, H. gelotopoeon, and C. virescens, and can also be used as a tool for monitoring the spread of these pests in agricultural regions.

Keywords

• molecular entomology, commodities
• cytochrome oxidase i
• helicoverpa
• nuclear elongation factor 1 alpha