A simply method for the addition of rotenone in Arabidopsis leaves.

MALIANDI, M. V.; RIUS, S.; BUSI, M. V.; DIEGO FABIAN GOMEZ CASATI

Rosario

Congreso; XLII Reunión SAIB; 2006

The NADH:ubiquinone oxidoreductase (complex I), a component of the respiratory chain, is a large multi-subunit complex and has been well characterized in bacteria, fungi, mammals and plants. Mechanistic studies showed that rotenone block the electron transport between a Fe-S cluster and ubiquinone. It has been described that several genes participating in protein import to mitochondria, molecular chaperones and genes involved in respiratory chain assembly are highly expressed after treatment of Arabidopsis cell cultures with rotenone. However, some methodological problems remain to be solved when used rotenone directly on Arabidopsis leaves instead of cell cultures. In the present work, we report a simple procedure for the treatment of Arabidopsis leaves with rotenone. The usefulness of the procedure was determined by the analysis of the expression two genes: COX19, involved in respiratory chain assembly and known to be induced in Arabidopsis cell cultures after rotenone treatment, and AOX, showing no alteration in the presence of rotenone. Results show that treated leaves have an increased expression of COX19 and no effects on AOX transcription. Moreover, rotenone/Triton X-100 incubated leaves presented an inhibition of oxygen uptake. The simplicity of the procedure shows this methodology useful for studying the effect of the addition of rotenone to a photosynthetic tissue in situ.