Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)

Abstract

Wolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity

Experimental hybridization and reproductive isolation between two sympatric species of tephritid fruit flies in the Anastrepha fraterculus species group

“…Such patterns could be explained by cytoplasmic incompatibility driven by the presence/absence of different strains of Wolbachia . In the fraterculus species group, A. ludens does not carry the bacterial symbiont (Martinez et al ., ), A. suspensa (Jeyaprakash & Hoy, ) and A. obliqua (Coscrato et al ., ) do, whereas cryptic species of A. fraterculus can display single and double strain infections (Selivon et al ., ; Cáceres et al ., ; Coscrato et al ., ; Marcon et al ., ). Novel strategies for area‐wide pest management take advantage of Wolbachia induction of cytoplasmic incompatibility for the development of the incompatible insect technique (IIT) (Sadiraki & Bourtzis, ).…”

Ongoing speciation within the Anastrepha fraterculus cryptic species complex: the case of the Andean morphotype

“…Some morphotypes of the A. fraterculus species complex have been found to carry single and even double infections of Wolbachia (Selivon et al, 2002;C aceres et al, 2009;Coscrato et al, 2009;Marcon et al, 2011), although several morphotypes have not been examined yet. They further conclude that a nuclear-cytoplasmic interaction may be at play because the cross of identical genotypes (F1 9 F1) did not result in reduced egg hatch.…”

“…We detected very similar patterns between the Andean morphotype and the four other A. fraterculus morphotypes (Mexican, Brazil-1, Brazil-3, Peruvian). Some morphotypes of the A. fraterculus species complex have been found to carry single and even double infections of Wolbachia (Selivon et al, 2002;C aceres et al, 2009;Coscrato et al, 2009;Marcon et al, 2011), although several morphotypes have not been examined yet. Post-zygotic isolation patterns in our study could be suggestive of cytoplasmic incompatibility due to Wolbachia, a hypothesis that could be corroborated by screening flies from the morphotypes evaluated here.…”

“…In general, heterotypic crosses in one male-female direction result in a reduction of F1 egg hatch, which vary in degree (Selivon et al, 1999(Selivon et al, , 2005C aceres et al, 2009;Rull et al, 2013) and some morphotypes have been found to carry Wolbachia infections (Selivon et al, 2002;C aceres et al, 2009;Coscrato et al, 2009;Marcon et al, 2011). In general, heterotypic crosses in one male-female direction result in a reduction of F1 egg hatch, which vary in degree (Selivon et al, 1999(Selivon et al, , 2005C aceres et al, 2009;Rull et al, 2013) and some morphotypes have been found to carry Wolbachia infections (Selivon et al, 2002;C aceres et al, 2009;Coscrato et al, 2009;Marcon et al, 2011).…”

“…All of the examined morphotypes display some degree of post-zygotic (postcopulatory) isolation. In general, heterotypic crosses in one male-female direction result in a reduction of F1 egg hatch, which vary in degree (Selivon et al, 1999(Selivon et al, , 2005C aceres et al, 2009;Rull et al, 2013) and some morphotypes have been found to carry Wolbachia infections (Selivon et al, 2002;C aceres et al, 2009;Coscrato et al, 2009;Marcon et al, 2011). Also, hybrid crosses in one direction between adults of some morphotypes have been found to produce an F1 female-biased progeny (Selivon et al, 1999).…”