2 n morpholino ethanesulfonic acid mes

Wild type A. thaliana seeds, ecotype Columbia, were kindly supplied by Prof. Dr. Richard Strasser (Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria). Seeds were surface-sterilized in 70% (v/v) ethanol for 2 min, followed by 8 min in 5% NaClO (Sigma-Aldrich, St. Louis, MI, USA). Afterwards, the seeds were rinsed eight times with sterile distilled water. The sterilized Arabidopsis seeds were sown in vitro on solid ½ Murashige and Skoog (MS) medium (2.154 g/L MS basal salt (Duchefa Biocheme, Haarlem, Netherlands), 10 g/L sucrose (Duchefa Biocheme), 0.1 g/L myo-inositol (Duchefa Biocheme), 0.5 g/L MES—2-(N-morpholino)ethanesulfonic acid (Roth, Karlsruhe, Germany), and 8 g/L plant tissue culture agar (Duchefa Biocheme), pH 5.7). After stratification for 3 days at 4 °C in the dark the plates were transferred to a growth chamber at 21 °C with a 16/8 h light/dark photoperiod for 3-4 days for root assays and 9 days for leaf assays. N. benthamiana seeds were kindly provided by Dr. Verne A. Sisson (Oxford Tobacco Research station, Oxford, NC, USA). N. benthamiana plants were sown in commercial soil in pots and kept at 28 °C under normal light conditions with a 16/8 h light/dark photoperiod.

Wild type Arabidopsis thaliana seeds, ecotype Columbia 0, were kindly supplied by Prof. Dr. Richard Strasser (Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria). When grown in vitro, seeds were surface sterilized in 70% (v/v) ethanol for 2 min, followed by 8 min in 5% (v/v) NaOCl (Sigma-Aldrich, St Louis, MO, USA). Afterwards, the seeds were rinsed eight times with sterile distilled water. The sterilized Arabidopsis seeds were sown in vitro on solid ½ Murashige and Skoog (MS) medium (2.154 g/L MS basal salt [Duchefa Biocheme, Haarlem, The Netherlands], 10 g/L sucrose [Duchefa], 0.1 g/L myo-inositol [Duchefa], 0.5 g/L MES - 2-(N-morpholino)ethanesulfonic acid [Carl Roth, Karlsruhe, Germany], 8 g/L plant tissue culture agar [Duchefa], pH 5.7) supplemented with 10 mg/L phosphinothricin (PPT, Duchefa) or 75 µg/ml kanamycin (Duchefa) as selective agents during selection. Alternatively, Arabidopsis seeds were grown in pots containing commercial soil or individually grown in artificial soil (Jiffy-7, 44 mm Ø). Both plants grown in vitro and in soil were first stratified for 3 days at 4°C in the dark and then transferred to a growth chamber at 21°C with a 16/8 h light/dark photoperiod.