Signalling events triggered in response to gibberellins during potato tuber sprouting

The aim of this study is to analize the genes that are up or down-regulated in response to sprouting promoting effectors such as GAs and light, in order to elucidate the signalling cascades triggered during this process. Half sections of certified pathogen tested Spunta tubers were put into sterile MS-agar (0.7%) containing flasks at 16„aC and subjected to different treatments: Dark controls: Tuber sections were incubated for 10 or 35 days in complete darkness. Light control: Tuber sections were incubated for 10 days under continuous light conditions. Treatments performed in complete darkness during 35 days: a) GA treatment: 5 uM gibberellic acid (C19H22O6) = GA3 (Sigma) b) Sucrose treatment: 1% sucrose c) GA & sucrose treatment: 5 uM GA3 (Sigma) + 1% sucrose Treatments performed in light (10 days)d) GA treatment : 5 uM GA3 (Sigma). Three independent biological replicates of each condition (3 controls and 4 treatments) wereperformed. At the moment of harvest (10 or 35 days) sprout length was measured and photographs of the different samples were taken. Sprouts were excised and frozen in liquid Nitrogen. Total RNA was obtained using the Trizol reagent (Invitrogen) according to the manufacturer¡¦s instructions. RNAs were quantified by OD: 260/280 and checked in agarose gels. Keywords: Loop design 25 hybs total