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

DREON, MARCOS SEBASTIÁN; CADIERNO, MARÍA PILAR; HERAS, HORACIO

Córdoba

Congreso; LII Reunión Anual de la Sociedad Argentina de Bioquímica y Bilogía Molecular; 2016

Universidad Nacional de Córdoba

The South American freshwater gastropod Pomacea canaliculata is a highly invasive species. It is a serious crop pest, being responsible for great economical losses and ecological damages. It is also a vector of a nematode that causes human meningoencephalitis. Its reproduction, among other aspects, has been extensively studied, but very little research has been conducted regarding gene expression. We performed the validation of reference genes, an essential pre-requisite for accurate qPCR studies that has not been determined for this species before, neither in any other freshwater gastropod. We selected the female albumen gland which plays a central role in reproduction and evaluated the expression stability of 7 candidate reference genes: EF1-a, RPL7, His H3.3, TUBB, 18S RNA, ACTB, and GAPDH. Stability was analyzed under different reproductive activity conditions employing three different approaches: geNorm, NormFinder, and the comparative ΔCt method. NormFinder analysis selects GAPDH and ACTB pair as the best option to be used for normalization, whereas geNorm and the comparative ΔCt method indicate RPL7, GAPDH, and 18S RNA as the most stable ones under the studied conditions. These results will be helpful to continue developing reproductive studies and particularly to perform future evaluation of the factors that may affect fecundity on this conspicuous invasive species by qPCR.