Validation by qPCR of reference genes for reproductive studies in the invasive apple snail Pomacea canaliculata

HERAS, HORACIO; HERAS, HORACIO; MARCOS SEBASTIAN DREON; MARCOS SEBASTIAN DREON; MARIA PILAR CADIERNO; MARIA PILAR CADIERNO

MALACOLOGIA

INST MALACOL

Lugar: Philadelphia; Año: 2018 vol. 62 p. 163 - 170

0076-2997

The South American freshwater gastropod Pomacea canaliculata is a highly invasive species.In introduced areas, it is a serious crop pest, responsible for great economic loss andecological damage. It is also a vector of the nematode Angiostrongylus cantonensis thatcauses human meningoencephalitis. Many aspects and particularly its reproduction havebeen extensively studied, but little research has been conducted on this species regardinggene expression. To meaningfully interpret quantitative PCR, a powerful technique to developthis kind of study, validation of reference genes is essential but until now has not been undertaken.We selected the female albumen gland for its major role in egg production in orderto evaluate the expression stability of the candidate reference genes EF1-α, RPL7, His H3.3,TUBB, 18S RNA, ACTB and GAPDH. Stability was analyzed under different reproductiveactivity conditions and defined based on three approaches: geNorm, NormFinder and thecomparative ΔCt method. NormFinder selected GAPDH and ACTB as the best option to beused for normalization, whereas geNorm and the comparative ΔCt method indicate RPL7,GAPDH, and 18S RNA as the most stable genes under the conditions studied. These resultswill facilitate reproductive studies, particularly those using qPCR to evaluate factors that mayaffect fecundity of this conspicuous invasive species.