Abiotic Stress Response in Arabidopsis

Arabidopsis WT seeds were surface-sterilized as described in [43 (link)]. The sterilized seeds were germinated and grown on a nylon screen placed on top of ½ strength solid MS media with vitamins (PhytoTechnology Laboratories, Lenexa, KS, USA), supplemented with 0.8% (w/v) Phytoblend agar (Caisson Laboratories, Smithfield, UT, USA), and 1% (w/v) sucrose, in Petri dishes for two weeks. Arabidopsis seedlings were exposed to 100 mM D-mannitol, 100 mM NaCl, and 2.5 μM ABA in liquid culture. Plant shoot and root samples were harvested separately at 0, 6, 12, and 24 h time intervals; washed with deionized water, flash frozen in liquid nitrogen, and stored at −80 °C until RNA extraction and qRT-PCR analyses.