In vitro culture is the major contributing factor for transgenic vs. non-transgenic proteomic plant differences

Fonseca, Cátia; Planchon, Sébastien; Serra, Tânia; Chander, Subhash; Saibo, Nelson; Renaut, Jenny; Oliveira, M.Margarida; Batista, Rita

http://hdl.handle.net/10400.18/1831

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Authors

Oliveira, M.Margarida

Batista, Rita

Abstract(s)

The identification of similarities and differences between genetically modified plants and derived food/feed and their comparators plays a central role in risk assessment strategy. Our main goal was to better understand the relevance of transgene presence, genetic and epigenetic changes promoted by transgene insertion and in vitro culture in the potential unintended effects/differences between transgenic and conventional counterparts. In order to achieve this goal we have used Multiplex fluorescence 2D gel electrophoresis technology coupled with MS to characterize the proteome of three different rice lines (Oryza sativa L. ssp. japonica cv. Nipponbare): a control conventional counterpart, an Agrobacterium-transformed transgenic line (Ta) and a negative segregant (NSb), progeny of a different transgenic line. We have observed that Ta and NSb plant lines grouped together (only 1 differentially regulated spot - fold difference ≥ 1.5), separated from the control (49 spots with fold difference ≥1.5, in both Ta and NSb lines comparing to control line). Since the only feature in common between the tested transgenic and negative segregant lines is that they have both suffered in vitro culture procedures we hypothesised that stress caused by in vitro culture was, in this case, the most relevant factor contributing for the encountered proteomic differences. MS identifications of differentially regulated spots supported our hypothesis, demonstrating that Ta and NSb rice lines present several adjusted metabolic pathways and several proteins with altered expression that were previously associated with plant stress response.