To understand mitogenome characteristics and reveal phylogenetic relationships of the genus Ostrinia, including several notorious pests of great importance for crops, we sequenced the complete mitogenomes of four species: Ostrinia furnacalis (Guenée, 1854), Ostrinia nubilalis (Hübner, 1796), Ostrinia scapulalis (Walker, 1859) and Ostrinia zealis (Guenée, 1854). Results indicate that the four mitogenomes-O. furnacalis, O. nubilalis, O. scapulalis, and O. zealis-are 15,245, 15,248, 15,311, and 15,208 bp in size, respectively. All four mitogenomes are comprised of 37 encoded genes and a control region. All 13 protein-coding genes (PCGs) initiate with ATN and terminate with TAN, with the exception of cox1 that starts with CGA, and cox1, cox2, and nad5 that terminate with an incomplete codon T. All transfer RNA genes (tRNAs) present the typical clover-leaf secondary structure except for the trnS1 (AGN) gene. There are some conserved structural elements in the control region. Our analyses indicate that nad6 and atp6 exhibit higher evolution rates compared to other PCGs. Phylogenetic analyses based on mitogenomes using both maximum likelihood (ML) and Bayesian inference (BI) methods revealed the relationship (O. palustralis + (O. penitalis + (O. zealis + (O. furnacalis + (O. nubilalis + O. scapulalis))))) within Ostrinia. Insects 2020, 11, 232 2 of 15East Asia, and Northwestern Africa. Within this third species group, three subgroups are recognized according to the differences in male mid-tibia. The first subgroup contains four species including O. nubilalis and O. furnacalis with smaller mid-tibia and lacking any groove or scales. The second subgroup includes two species, namely O. kurentzovi (Mutuura and Munroe, 1970) and O. narynensis (Mutuura and Munroe, 1970), with a moderately dilated tibia and a fringe of enlarged curved scales. The third subgroup consists of four species, including O. scapulalis and O. zealis, possessing a strongly dilated tibia and massive scales in the groove. Mutuura and Munroe (1970) stated that the three species groups within the genus Ostrinia are monophyletic and indicated that O. penitalis is the most primitive species. The second group was defined as monophyletic because the members share potential synapomorphies in male genitalia [2]. The third species group has been intensively studied on the basis of their morphology, pheromones, and DNA sequence. Based on the complexity of the male tibia, Mutuura and Munroe (1970) considered that the species with the smallest tibia is the most primitive, then the species with the medium tibia are intermediate, and finally, the species with the large tibia and massive tuft are the most derived.Frolov (1981, 1984) stated that the variation in the male mid-tibia was determined by two alleles. The Mt and Mt + were two alleles located on autosomes and controlled the size of the mid-tibia, while i and i + were two alleles located on Z-sex-linked chromosomes and controlled the groove appearance. Mt/Mt + and i/i + follow a Mendelian inheritance pattern ...

Nagiella occultalis Misbah & Yang, sp. n. from China is described and illustrated. This new species is very similar to N. quadrimaculalis (Kollar, 1844) in general morphological characters of forewing and male genitalia. Molecular evidence shows that these two species diverge in COI barcode region by more than 3.2%. Sequence divergence among the two species is congruent with subtle morphological differences. Wing venation and male genitalia of the two species are compared and illustrated.

Integrative taxonomic study of three species of the genus Tylostega revealed that the genetic distances of the COI gene among the tested species was relatively large (3.27-7.60%). The Automatic Barcode Gap Discovery (ABGD) system performed better than the Barcode Index Number (BIN) in discriminating closely related species. This work provides a molecular baseline for future integrative taxonomic study of Crambidae.