Ms basal salt medium

Nicotiana tabacum plants (cv. Petit Havana SR1) as well as transgenics were cultured in sterile conditions on Murashige and Skoog (MS) basal salt medium (Duchefa Biochemie, Haarlem, the Netherlands) containing 0.8% plant agar (Duchefa Biochemie) or in soil in a growth room at 25°C with a 16-h/8-h light/dark cycle.

Seeds were sterilized, stratified for 2–3 days at 4 °C, and plants were grown on plates on solid 0.5× Murashige and Skoog (MS) basal salt medium (Duchefa, Brussels, Belgium). Plants were analyzed on plates or transferred to soil (“Einheitserde,” Klasman, Switzerland) 10 days after germination. Alternatively, seeds were directly sown on soil. Plants were grown in Conviron growth chambers with mixed cold fluorescent and incandescent light as indicated for each experiment (80 or 130 μmol m^-2 s^-1, 21 ± 2°C) under long day (LD, 16 h/8 h day/night cycle) or short day (SD, 8 h/16 h day/night cycle) photoperiods. Flowering time was scored as described (Möller-Steinbach et al., 2010 (link)). Each presented flowering time analysis represents one out of at least two performed experiments.