Salinity Stress Response in Camelina

Sterilized camelina seeds were planted at a depth of 2 cm in pots containing peat moss and were kept under the conditions of 16 h of light and a temperature of 25 °C with irrigation every three days. Then, the five-week-old seedlings were treated with salt (200 mM of NaCl) via irrigation, which was repeated after 24 h. After the salt treatment, leaves were collected at different time points (after 6, 24, and 72 h). The total RNA samples were extracted using an RNX kit (Sinaclon, Iran) and the cDNA was synthesized using a reverse transcriptase kit (Roche, Germany), according to manufacturer protocols. In the present study, five members of the SULTR family were selected for real-time PCR analysis. The genes were selected based on the phylogeny analysis. In addition, actin-2 (Csa15g026420) was used as a reference gene to normalize the expression data. Specific primers were designed using the Primer3 online software (version 4.1.0) [72 (link)], based on the coding sequences of the selected SULTR genes (Table S3). The expression patterns of the SULTR genes were analyzed using a Maxima SYBR Green/ROX qPCR Master Mix kit (Thermo Fisher, France) and the ABI Step One, according to manufacturer protocols. The expression levels of each SULTR gene were calculated using the delta Ct method [73 (link)], using three biological replicates.