Caracterização de ovos e larvas de espécies de Anastrepha (Diptera: Tephritidae) utilizando análises morfológicas e moleculares

Abstract

The identification of species of Anastrepha Schiner (Diptera: Tephritidae) is based on the shape of the female´s aculeus. However, it is the immature stages that are commonly intercepted at ports of entry. Thus, the correct identification of immature stages is highly desirable. This study aimed at contributing to the identification of the immature stages of Anastrepha species using a morphological and a molecular approach. The morphology of eggs of 11 species and larvae of eight species of Anastrepha was described in details for the first time. Diagnostic characters that can distinguish among the eggs of the 11 species include chorion ornamentation; location, presence or absence of aeropyles; location of the micropyle; pronounced rim of the chorion surrounding the micropyle and presence or absence of a respiratory appendage. The eggs of Anastrepha montei Lima e Anastrepha curitis Stone have a narrow patch of seemingly porous surface of the anterior pole. This character is reported for tephritid for the first time. Diagnostic characters that can distinguish among the larvae of the eight species studied include presence or absence of dorsal spinules on segments thoracic and abdominal, number of ventral spinules on the segments thoracic and abdominal, shape of the anal lobe, number of the posterior spiracular hairs and some characteristics of the cephalopharyngeal skeleton such as total length, size and shape of the mandible. PCR-RFLP (polymerase chain reaction associated to restriction fragment length polymorphism analysis) was used for the molecular characterization. Two regions of the mitocondrial DNA (COI and 16S) and one region of the nuclear DNA (ITS1) were screened using six restriction enzymes to screen 13 Anastrepha species. Interesting results to distinguish among species were obtained for the fragments 16S and ITS1 for Anastrepha atrigona Hendel, A. curitis, Anastrepha manihoti Lima, Anastrepha pulchra Stone, Anastrepha striata Schiner, Anastrepha sororcula Zucchi, and Anastrepha zenildae Zucchi. New associations of fruit flies with their hosts and parasitoids were reported for the first time.