PCR–RFLP identification of endemic Chilean species of Rhagoletis (Diptera: Tephritidae) attacking Solanaceae

Abstract

Four species of the genus Rhagoletis are native to Chile: R. nova (Schiner), R. conversa, (Brèthes), R. penela Foote and R. tomatis Foote. Currently, identification of these species is based on morphological criteria, but their strong similarity makes precise recognition difficult. To clarify species separation for quarantine purposes, a reliable method based on a PCR-RFLP procedure is reported. A DNA region containing mitochondrial NADH dehydrogenase genes was selected as a target sequence for the analysis. The amplification products (c. 1 kb) were digested with either SspI or DdeI, yielding specific patterns that differentiated each of the endemic species. Complete nucleotide sequences were determined, confirming empirical restriction maps. This report updates information on the geographical distribution of Rhagoletis species in Chile.

Phylogeographical analysis of neotropical Rhagoletis (Diptera: Tephritidae): did the Andes uplift contribute to current morphological differences?

“…8), and the close phylogenetic affi nity found herein, support the notion that these species have undergone a recent divergence. Because different host plants have been described for these species, namely Solanum muricatum (Aiton) for R. nova, and Physalis sp., Solanum tomatillo and Solanum nigrum for R. conversa (Foote 1981;Frías 1992Frías , 2001Salazar et al 2002), a process of sympatric speciation by host shift could have promoted this divergence.…”

“…Bands were excised from gels and DNA was extracted using a Wizard PCR purification kit (Promega, USA). Purifi ed PCR products were ligated into the pGEM5-T cloning vector (Promega, USA) as described by Salazar et al 2002. DNA sequencing of the inserts in pGEM5-T was performed by the Departamento de Ciencias Ecológicas, Pontifi cia Universidad Católica de Chile.…”

DNA Diagnostics to Identify Internal Feeders (Lepidoptera: Tortricidae) of Pome Fruits of Quarantine Importance

“…Most DNA-based diagnostics for quarantine applications in arthropods have used restriction enzyme digestion of PCR products generated by ampliÞcation with universal primers (PCR-restriction fragmentlength polymorphism) (Armstrong et al 1997(Armstrong et al , 2003Lee and Lee 1997;Salazar et al 2002). PCR with species-speciÞc primers allows diagnosis in a shorter time by avoiding the 2-h or more incubation time required for DNA digestion.…”

“…Examples include Tetranychus mites (Lee and Lee 1997), Bactrocera fruit ßies (JeomHee et al 2000), Liriomyza leafminers (Scheffer et al 2001), and tortricid leafrollers (Sperling and Hickey 1995). Other mitochondrial genes also have proven to be useful, such as NADH in Rhagoletis fruit ßies (Salazar et al 2002). Most diagnostic protocols for arthropods have relied on restriction analysis of a PCR product (PCR-restriction fragment-length polymorphism) ampliÞed using universal primers (Simon et al 1994).…”

Fruit Flies (Diptera: Tephritoidea): Biology, Host Plants, Natural Enemies, and the Implications to Their Natural Control

“…In the Brazilian territory are reported three species (Ragoletis adusta Foote, from the state of São Paulo, R. ferruginea Hendel, in Bahia, Paraná, and Santa Catarina, and R. macquarti (Loew), in Goiás, and Minas Gerais (Foote, 1981;Ramírez et al, 2008), but the species of Rhagoletis have not been considered as key pests in Brazil. On the other hand, some species in this genus are pest of fruits in Peru and Chile (Salazar et al, 2002).…”