In all experiments seeds of T or T plants were

In all experiments, 10 seeds of T2 or T3 plants were grown vertically in sterile square Petri image dishes (Corning, 431.301; 20 cm × 20 cm) under controlled conditions (day/night temperature of 28/25 °C, K-Ras(G12C) inhibitor 6 12 h photoperiod, and a light intensity of 500 ?Em−2 s−1) as described previously [52]. Briefly, after sterilization, the seeds were sown on square Petri dishes containing 250 mL of half strength Murashige and Skoog (MS/2) solid medium with the radicle oriented downwards. The MS/2 solid medium was composed of 2.15 g L−1 of Murashige and Skoog medium basal salt mixture (Duchefa Biochemie, M0221), 75 mg L−1 Murashige and Skoog vitamin mixture (Duchefa Biochemie, M0409) and 8 g L−1 of agarose type II (Sigma–Aldrich, A6877). For salt and mannitol medium, 7 g L−1 of NaCl (120 mM) and 21.9 g L−1 of mannitol (120 mM), respectively, were added to MS/2 medium before autoclaving. After 6 days of growth, the Chromatin remodeling lengths of the seminal root and second leaf (i.e., the leaf following the first incomplete leaf) were recorded for each of the 10 plantlets.
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