growth-protocol.md

Arabidopsis thaliana Heynh. Col–0 and its mutant bou‒2 (Li et al., 2003; Rosso et al., 2003), available from the Nottingham Arabidopsis Stock Centre as N370142 and harbouring a T-DNA insertion in the second exon of the BOU gene sequence, were used in this study. For sterilization, Arabidopsis seeds were washed twice with 70% (v/v) ethanol, containing 1% (v/v) Triton X‒100, twice with 100% ethanol and dried. Seeds were grown on 0.5 Murashige and Skoog (MS) medium (Murashige & Skoog 1962) with 0.8% (w/v) agar at pH 5.8. After cold stratification at 4°C to synchronize germination, seedlings were grown at 60% relative humidity, 100 – 150 µmol photons m-2 s-1, 12 h light / 12 h dark, 22°C / 18°C in a 3,000 ppm CO2-enriched atmosphere. Seedlings with two developed leaves were transferred to soil and cultivated under the conditions indicated above.