Illumina HiSeq 2000 paired end sequencing; Transcriptional response in leaves and root of potato plants to cadmium (Cd) exposure
Transcriptional response in leaves and root of potato plants to cadmium (Cd) exposure
Youngest leaves from the third row of the canopy were collected and flash frozen in liquid nitrogen and stored at -80 until further processing. Virus-free tubers of the variety Rooster were sprouted under constant diffused light conditions and planted in 10 L pots filled with a sand-soil mixture under controlled greenhouse conditions with 16 h light at 210C. The plants were watered from above daily with 1 L nutrient solution as described in Dunbar et.al (2003). Briefly, the nutrient solution comprised (mM) 3.0 KNO3, 1.0 MgSO4, 0.5 NaCl, 2.5 CaNO3, 0.5 K2SO4, and 0.5 KH2PO4. Micronutrients (in μM) were 20 B, 20 Mn, 10 Zn, 1.0 Cu, and 0.5 Mo. Iron (25 mM) was added as a complex with DTPA. The nutrient solution was adjusted to a pH of 5.5 with HCl.  During the growth period, the plants were rotated every day to have uniform light exposure. At 45 days of planting, the plants were randomly assigned into one of three treatments. Plants in the first treatment did not receive any Cd and acted as the experimental control, plants in the second treatment received 1 mg kg-1  Cd for 72 hours, and plants in the third treatment received 5 mg kg-1 Cd for 72 hours.  Cd was applied in the form of CdCl2 with the nutrient solution.  Total RNA extraction from leaf and root tissue was performed using Trizol reagent.  TruSeq RNA sample Prep Kit (Illumina, San Diego, CA, USA)