Cenchrus americanus (genotype ICMR 1122) plants treated with Low VPD conditions
Total RNA was extracted from root tissues sampled from PM plants subjected to high and low VPD conditions by using 100 mg of tissue with RNeasy Plant mini kit (Qiagen, Germany), according to manufacturers instructions. The integrity of the RNA was checked by performing agarose gel electrophoresis and using BioAnalyzer (Agilent). sRNA fractions (~18 to 30 nt) were purified by running total RNA on a 15% polyacrylamide gel and eluting the portion corresponding to small RNAs. The obtained small RNA was sequentially ligated to 3 and 5 adapters using T4 RNA ligase 1 (NEB). The adapter-ligated sRNAs were used for cDNA synthesis using ProtoScript kit (NEB). The cDNA was further purified and subjected to sequencing using Illumina MiSeq platform. Adaptor trimming was performed, and low-quality reads were removed, and high-quality reads were further subjected downstream analysis