DesC_3
Cicer arietinum
Protocols: The leaves of 12 chickpea plants (3 biological replicates for each of the 2 genotypes and 2 water regimes) were collected, immediately frozen in liquid nitrogen and stored at −80 °C The two chickpea genotypes were used: Desi PI598080 and Kabuli Flip07 318C, respectively sensitive and tolerant to drought stresses. Seeds were planted in pots filled with potting soil (horticultural soil of the COMPO SANA® company). On the 21st day after sowing (DAS) the seedlings were thinned to one healthy plant per pot. The plants were well-watered until the beginning of the treatment. After 12 weeks from the germination, pots were separated into two groups: the first group was watered to 70% of the evapo-transpired (EVT) water (control), while the second group was watered to 30% of the EVT water (water stress conditions). The experiment was conducted in a greenhouse under natural light. The average daily temperature inside the greenhouse was the following: 21°C when the plants germinated and 26° C when the stress started. Plants were fertilized with commercial fertilizer (NPK 7:7:7) once a week. Frozen leaves (100mg) have been grounded to a fine powder with a previously sterilized mortar and pestle using liquid nitrogen to prevent RNA degradation. RNA extraction was performed using 100 mg of grounded material using the RNeasy Plant Mini kit (Qiagen). Final elutions were performed using 60 μL of RNase-free water. RNA quality was determined using the Agilent 2100 Bioanalyzer (RNA 6000 nano kit - Agilent Technologies). Twelve transcriptomic libraries were prepared using Illumina mRNA Prep kit, following the manufacturer instructions and a unique dual index combinations were used for each sample for barcoding. The concentration of each library was determined using Qubit™ 4 Fluorometer (dsDNA High Sensitivity Kit - Invitrogen).