Investigating the role of genes expressed in male reproductive tissues of Anastrepha obliqua on their reproductive success

Abstract

The West Indies fruit fly Anastrepha obliqua, as well as all other flies of the Tephritidae family, requires fleshy fruits during part of his life cycle to oviposit and serve as food for their larvae, rendering these fruits unviable for human consumption. This species is a significant pest in the cultivation of fruits, causing great damage because of its diverse host range and broad distribution throughout the American continent,. One of the existing methods for pest control is the Sterile Insect Technique (SIT) in which sterile males are released in the infested environment and compete with wild males, decreasing the amount of reproductive success and consequently the fly population of the next generation. The most widely used method for making sterile males is the use of radiation, but the side effects of this technique often make males less competitive. One way that this technique could be improved is by the use of genes related to sexual behavior that may be manipulated to make the male sterile while still competitive. In order to search for such genes we investigated differentially expressed genes in reproductive tissue of virgin males and post-copulatory males using next generation sequencing. We chose three genes: Attacin A, Obp56 and trypsin theta which will be investigated by real-time PCR (qPCR) for differential expression in different stages and tissues and select one of them that shows a promising differential expression pattern that might make it amenable to be considered to be silenced by RNA interference (RNAi). We will use qPCR and fertility tests and measurements to verify the efficiency of the silencing and its effects on the phenotype.