Identification of Diatraea 1 spp. (Lepidoptera: Crambidae) based on Cytochrome oxidase II

Diatraea spp. (Lepidoptera: Crambidae) are a group of insects that are agriculture pests in 17 many economically relevant crops such as sugarcane, sorghum, corn and rice. Recognized 18 species for this genus respond differentially to natural enemies used in their biological 19 control, emphasizing the importance of species in a regional approach. Currently, 20 identification is based on the male genitalia. However, the availability of specimens collected 21 from field and subjectivity based on the character recognition can seriously hamper species 22 identification, and therefore result in inadequate pest management. To overcome this, 23 individuals of Diatraea spp. preliminarily classified male genitalia and obtained from reared 24 conditions and the field (both derived from natural populations occurring in Colombia) were 25 analyzed using genitalic morphometry and molecular biology specifically using a fragment 26 of the cytochrome oxidase subunit II (CO II) mitochondrial gene. Although morphometric 27 analysis did not show any overriding results regarding genitalia morphology, the 28 bioinformatics analyses of CO II sequences resulted in an adequate classification of the 29 individuals within the recognized species. It also, revealed that the occurrence of clades 30 associated with geographical distribution may be associated with cryptic species. The latter 31 was also confirmed by a Single-Strand Conformation Polymorphism (SSCP) methodology 32 evaluating the same fragment of CO II. This experimental approach allows properly 33 recognizing each species and in consequence is proposed as an effective tool in Diatraea 34 species identification.