Identification of suitable reference genes for real-time RT-PCR normalization in Bidens laevis during pesticide and cold stress.

LUKASZEWICZ GERMÁN; AMÉ, MARÍA VALERIA; MENONE, MIRTA LUJÁN

Cracovia

Encuentro; 3rd Young Environmental Scientists Meeting; 2013

SAC-SETAC

Environmental monitoring demands the use of new species and sensitive methods in order to get accurate regional biomonitoring. The aquatic macrophyte Bidens laevis L. (fam.: Asteraceae) has desirable growing characteristics for laboratory assays, has proven to respond positively to genotoxic compounds and showed enzymatic response when exposed to the pesticide endosulfan. Nowadays, gene expression offers crucial information when analysing responses to xenobiotic compounds. It is well known that Real Time PCR (RT-qPCR) is the preferred method for studying gene expression because of its sensitivity, precision and robustness but it requires the use of reference genes (RG) with stable expression among different treatments, as well as different tissues and growth stages. The aim of this study was to analyse the expression of six candidate genes, Elongation Factor 1-alfa (EF1a), Beta-Actin (BACT), Histone 3 (H3), Nicotinamide Adenine Dinucleotide Dehydrogenase (NADHD) and glyceraldehyde-3-phosphate-dehydrogenase (GADPH), in B. laevis in three tissues (root, stem and leaves) under four conditions: 1-control-: Large plants (>1500mg), temperature 22ºC, media: Hoagland solution; 2 -short-: Short plants (